MODEL U-5100 RATIO BEAM SPECTROPHOTOMETER

Please read through this manual carefully

and keep it in a safe place.

• Before using the instrument, read the safety

instructions and precautions carefully.
• Keep this manual in a safe place nearby so it can
be referred to whenever needed.

HITACHI
 INSTRUCTION MANUAL

FOR

MODEL U-5100 RATIO BEAM SPECTROPHOTOMETER

Hitachi High-Technologies Corporation

Copyright C Hitachi High-Technologies Corporation 2010. 1st Edition, 2010

All rights reserved. Printed in Japan. Part No. 3J2-9001 HT-F (H-LT)
Be sure to read through and understand the following points
with regard to this manual.

1. Information contained in this document is subject to

change without notice for improvement.
2. This manual is copyrighted by Hitachi High-Technologies
Corporation with all rights reserved.
No part of this manual may be reproduced, transmitted
or disclosed to a third party in any form or by any means
without the express written permission of Hitachi
High-Technologies Corporation.
3. Hitachi High-Technologies Corporation assumes no
liability for any direct, indirect, or consequential damages
arising from use not described in this manual.
Utmost care must be exercised when using the
instrument.
4. This document does not provide any warranty or
permission for industrial properties or any rights to grant
license lawfully and without infringement.
PREFACE
We thank you for purchasing the Model U-5100 Ratio Beam
Spectrophotometer.

This instrument is designed for measuring the absorbance and

transmittance of a sample.
Note that samples that may have been infected with bacteria or
viruses are not applicable to the instrument

This instrument is intended for use by persons having a

basic knowledge of chemical analysis procedures.
Keep in mind that improper use of analytical instruments,
chemicals or samples would result not only in wrong
analytical data but also in consequences adverse to safety.
Note that it is allowed only for persons having a basic
knowledge of chemical analysis procedures to use this
instrument.

Please read this instrument manual carefully before attempting

operation and acquaint yourself with this instrument for its
correct use.

ABOUT THIS MANUAL

This instruction manual has been prepared for the user of Model
U-5100 Ratio Beam Spectrophotometer. The operating
procedures and maintenance/inspection instructions for the
instrument are contained in this manual.

First of all, be sure to read “IMPORTANT” and “SAFETY

SUMMARY” at the beginning of this manual.

The contents of “IMPORTANT” and “SAFETY SUMMARY”

described hereafter apply to the accessories of this instrument
also.
IMPORTANT

Precautions on CE Conformity Marking

In consideration of use in the European countries, this instrument

bears the CE mark indicating the conformity to the requirements
mentioned below.

1. Electromagnetic Compatibility Requirement

This instrument is designed to satisfy the European Norm

EN61326-1 (2006) for the CE conformity marking through
conformity to the EMC Directive 2004/108/EC.

This instrument is classified as Class A of EN61326-1. So, this

instrument must not be used in domestic establishments nor in
establishments directly connected to a low voltage power supply
network which supplies buildings used for domestic purpose.

And this instrument is also designed to comply with table 1

"Basic immunity test requirements" in the above European
Norms. If the instrument is used near an intense
electromagnetic source, however, interfering noise may be given
to the instrument to cause an adverse effect on its performance
or functionality.

2. Safety Requirement

This instrument is also designed to satisfy the European Norm

EN61010-1 (2001) for the CE conformity marking through
conformity to the LVD Directive 2006/95/EC.
This instrument is requested to be used in a suitable
environment and grounded appropriately.

IMPORTANT - 1
Information for Users on WEEE (only for EU Countries)

This symbol is in compliance with the Waste Electrical and

Electronic Equipment directive 2002/96/EC (WEEE).
This symbol on the product indicates the requirement NOT to
dispose of the equipment as unsorted municipal waste, but use
the return and collection systems available.

Information on Disposal for Users

1. In the European Union

If you need to discard this product or discard user serviceable

parts:
Please contact your local sales representative or distributor who
will inform you of the recycle of the product.
You might be charged for the costs arising from take-back and
recycling.

2. In other Countries outside the EU

If you wish to discard this product, please contact your local

authorities and ask for the correct method of disposal.

IMPORTANT - 2
Warranty on Product

This product, inclusive of its accessories, is warranted to be free

from defects in material or workmanship under normal use within
the product specifications indicated in this manual and under
conditions given below.
This warranty is void if the instrument is not used according to
the instruction manual.

(1) Scope of Warranty

Any parts that prove to be defective in design or

workmanship during the warranty period will be repaired,
adjusted or replaced without charge. A substitute part
may be used for repair, or replacement with an equivalent
product may be made instead of repair. Such system
components as a personal computer and printer to be
updated frequently for improvement may not be available
in original versions at the time of replacement.
The manufacturer assumes no liability for any damage to
data or application software due to any possible fault or
failure of this instrument.

(2) Warranty Period

One year from the date of initial installation

(In case a separate warranty document has been issued,
the warranty period indicated in it takes precedence over
the above period)

IMPORTANT - 3
(3) Limitations and Exclusions on Warranty

Note that the following cases are excluded from the scope
of this warranty, i.e., these cases are beyond the coverage
of free-of-charge repair even during the warranty period
indicated above.

(a) Failure due to operation at a place not meeting the

installation requirements specified by the
manufacturer.

(b) Failure due to power supply voltage/frequency other

than specified by the manufacturer or due to
abnormality in power supply.

(c) Corrosion or deterioration of the piping due to

impurities contained in gas, compressed air or
cooling water supplied by the user.

(d) Corrosion of the electric circuits or deterioration of

the optical elements due to highly corrosive
atmospheric gas.

(e) Failure due to use of software, hardware or spare

parts not supplied by the manufacturer.

(f) Failure due to use not described in the manual or

improper repair not approved by the manufacturer.

(g) Failure due to maintenance or repair by other than

service engineer qualified by the manufacturer.

(h) Failure due to relocation or transport conducted not

under the supervision of the manufacturer after the
initial installation of the instrument.

(i) Failure due to disassembly, modification or relocation

not approved by the manufacturer.

(j) Failure due to acts of God, including fire, earthquake,

storm, flood, lightning, social disturbance, riot, crime,
insurrection, terrorism, war (declared or undeclared),
radioactive pollution, contamination with harmful
substances, etc.

IMPORTANT - 4
 (k) Failure of the hardware, or damage to the system
software, application software or data due to
computer virus infection.

(l) After disposal of this instrument, after its resale

without prior approval from the manufacturer,
consumable parts, and failure of any part that have
reached the end of its service life.

(m) Failure due to a life-limited part that has exceeded

the end of its useful lifetime.

(4) Disclaimer of Warranty

THE MANUFACTURER MAKES NO WARRANTIES,

EITHER EXPRESS OR IMPLIED, EXCEPT AS
PROVIDED HEREIN, INCLUDING WITHOUT LIMITATION
THEREOF, WARRANTIES AS TO MARKETABILITY,
MERCHANTABILITY, FOR A PARTICULAR PURPOSE
OR USE, OR AGAINST INFRINGEMENT OF ANY
PATENT. IN NO EVENT SHALL THE MANUFACTURER
BE LIABLE FOR ANY DIRECT, INCIDENTAL OR
CONSEQUENTIAL DAMAGES OF ANY NATURE, OR
LOSSES OR EXPENSES RESULTING FROM ANY
DEFECTIVE PRODUCT OR THE USE OF ANY
PRODUCT.
NO ORAL OR WRITTEN INFORMATION OR ADVICE
GIVEN BY THE MANUFACTURER, ITS DEALERS,
DISTRIBUTORS, AGENTS OR EMPLOYEES SHALL
CREATE A WARRANTY OR IN ANY WAY INCREASE
THE SCOPE OF THIS WARRANTY.

IMPORTANT - 5
Installation, Relocation and After-sale Technical Service

(1) Installation and Relocation

(a) Installation at delivery shall not be carried out by the

user. It shall be carried out by our sales
representative or the engineers who have been
trained and qualified for this purpose by us in order to
use the instrument safely and accurately.

(b) Before installation, the user shall make preparations

for satisfying the installation requirements in
accordance with this instruction manual.

(c) If relocation becomes necessary after initial

installation (delivery), please contact the dealer from
whom you purchased the instrument or our sales
representative.

(2) After-sales Service

(a) For after-sales service, contact our sales

representative or service office of our sales
representative.

(b) For service after the warranty period, consult our

sales representative or service office of our sales
representative with regard to a maintenance and
inspection service contract.

(c) The maintenance parts and consumables of the

instrument will be provided for 10 years after its
delivery. If a part or unit is unavailable due to the
discontinuance of its manufacture, a substitute part
or unit may be supplied, for which we request your
understanding. Even after the instrument has
reached its useful life, the parts and units will be
supplied where they are available, but this will not
assure an extension of the useful life.
In the case that the maintenance parts and
consumables are unavailable after a lapse of 10
years, please stop using the instrument and replace
the instrument with a new one.
Gas leak, water leak, electrical leak or ignition may
be caused if the instrument is used without
maintenance.

IMPORTANT - 6
 (d) Gas leak, water leak, electrical leak or ignition may
be caused if the maintenance parts and consumables
of the instrument is used without replacement.

(e) It may be impossible to supply the main unit

components other than the maintenance parts and
consumables due to the discontinuance of main unit
manufacture, etc. If the instrument becomes faulty,
it might be irreparable due to lack of such
components. In this case, please stop using the
instrument.

Technical Seminars and Training Courses for Users

We offer technical seminars and training courses at either our or

user’s facilities to ensure proper and safe operation of the
analytical instrument to its full performance. For further
information, contact our sales representative. (Applicants will
be charged.)

IMPORTANT - 7
Service Life of This Instrument

This instrument has a useful service life of seven years after the
date of its initial use (installation), which is estimated under the
condition that periodic maintenance, checkup, replacement of
life-limited parts, and repair of worn parts are carried out as
specified in the present instruction manual.
(In use of the instrument under standard operating conditions
(8h/day, 20 days/month))

Gas leak, water leak, electrical leak or ignition may be caused if

the maintenance parts and consumables of the instrument is
used without replacement.
Gas leak, water leak, electrical leak or ignition may be caused if
the instrument is used beyond the useful service life.

Other Precautions

1. Handling of Chemicals and Samples

(1) The user is responsible for following relevant legal

standards and regulations in handling, storage and
disposal of chemicals and samples used in analytical
operations with this instrument.

(2) Reagents, standard solutions and accuracy-control

samples shall be handled, stored and discarded as
instructed by the respective suppliers.

(3) Samples that may have been infected with bacteria or

viruses are not applicable to the instrument.)

2. Trademark Acknowledgments

Microsoft, Windows, Microsoft Excel, Microsoft Word, and

Windows XP are registered trademarks of Microsoft Corporation
of the USA. All other brand names and product names used in
this manual are registered trademarks or trade names of their
respective holders.

IMPORTANT - 8
 SAFETY SUMMARY
General Safety Guidelines

Before using Model U-5100 Ratio Beam Spectrophotometer, be

sure to read the following safety instructions carefully.

The hazard warnings which appear on the warning labels on the

product or in the manual have one of the following alert headings
consisting of a safety alert symbol and signal word DANGER,
WARNING or CAUTION.

: Safety alert symbol used for calling

attention to a potential hazard which
could cause personal injury.
To avoid possible injury or death,
observe all the safety messages
following this symbol.

DANGER : Indicates an imminently hazardous

situation which, if not avoided, will result
in death or serious injury.

WARNING : Indicates a potentially hazardous

situation which, if not avoided, can
result in death or serious injury.

CAUTION : Indicates a hazardous situation which, if

not avoided, can result in minor or
moderate injury.

NOTICE : Indicates a hazardous situation which, if

not avoided, can result in damage to
property.

In addition to the above, the following signal word is used to

indicate instructions for ensuring proper use of the product.

NOTE: Indicates an instruction for ensuring correct use

of the product and accurate analysis therewith.

SAFETY - 1
 SAFETY SUMMARY
Common Safety Precautions

Prior to Use

• Before using the instrument, be sure to read this instruction

manual carefully to attain a full understanding of its operations.

• Keep the instruction manual handy nearby so it can be

referred to whenever needed.

• Be sure to observe the procedures specified in the manual.

• Be sure to understand and follow all the safety instructions

given in the manual.

• Be sure to observe all the hazard warnings attached to the

instrument or provided in the manual. Failure to do so could
result in personal injury or damage to the instrument.

• Be sure to follow all the methods of use instructed in the

manual for proper application of the product.

• Absolutely avoid modifying the product, using non-specified

parts, or removing safety devices as it could be hazardous.

• Do not perform any operation or action other than described

in the manual.
On occurrence of any trouble in the instrument, notify your
nearest Hitachi High-Technologies Corporation sales
representative or service office of Hitachi High-Technologies
Corporation sales representative.

• When using chemicals for the instrument, be sure to provide

proper ventilation of the room. Inadequate ventilation could
endanger human health.

• Keep in mind that the hazard warnings in the manuals or on

the product cannot cover every possible case, as it is
impossible to predict and evaluate all circumstances
beforehand. Always be alert and use your common sense.

SAFETY - 2
 SAFETY SUMMARY
Common Safety Precautions (Continued)

• Wear appropriate protective equipment when using chemicals.

In the case of accidental contact with the skin and ingestion,
refer to the product safety data sheet to take first-aid action
and seek medical care.

In Use

If an abnormality such as unusual noise, odor, fuming or gas

leakage occurs during operation of the instrument, immediately
disconnect power to the instrument, and take proper safety
measures as required. Then, notify your nearest Hitachi High-
Technologies Corporation sales representative or service office
of Hitachi High-Technologies Corporation sales representative.

Installation, Maintenance, and Relocation

• At the time of delivery, installation of the instrument shall be

carried out by or under supervision of qualified service
personnel of the manufacturer or its authorized service agent
for ensuring safety and high accuracy in operation of the
instrument. It is not permitted for the user to carry out
installation.

• After completion of installation, check that all the standard

parts are equipped. If the instrument is made active with any
one of the standard parts not equipped, a failure could occur
to result in a hazardous condition.
If any item is missing or damaged or if you have any question,
notify the installation personnel at site or your nearest Hitachi
High-Technologies Corporation sales representative or
service office of Hitachi High-Technologies Corporation sales
representative.

SAFETY - 3
 SAFETY SUMMARY
Common Safety Precautions (Continued)

• The maintenance and checkup procedures to be taken by the

user are only those described in the manual. When taking
the maintenance and checkup procedures described in the
manual, attain a clear understanding of them.
Do not perform other maintenance and checkup procedures
to avoid jeopardizing safety and causing troubles in the
instrument.

• After installation, do not relocate the instrument. If the

instrument is relocated, vibration or impact to be applied
during relocation could cause a malfunction in the optical
components that have been adjusted precisely.

• If any warning/caution label has become illegible due to

deterioration with age or it has been damaged due to any
cause, notify your nearest Hitachi High-Technologies
Corporation sales representative or service office of Hitachi
High-Technologies Corporation sales representative for
replacement with a new one.

• Replace life-limited parts of the instrument with new ones at

intervals specified in this manual. Otherwise, a failure could
occur to result in gas leak, water leak, electrical leak or
ignition.
For other than the replacement procedures instructed in this
manual, contact your nearest Hitachi High-Technologies
Corporation sales representative or service office of Hitachi
High-Technologies Corporation sales representative.

SAFETY - 4
 SAFETY SUMMARY
Safety Instructions in This Manual

Shown below are the safety instructions contained in this manual

and their relevant sections in it.

DANGER Indications

The indication” DANGER” does not apply to this instrument.

WARNING Indications

Electric Shock due to Contact with Dangerous Voltage

• In contact with power supply voltage, you may receive an

electric shock to cause fatal or serious injury.
Before connecting the power cord, make sure that the
POWER switch of the spectrophotometer main unit is turned
off. Absolutely avoid modifying the product.
(Sections 1.5)

Electric Shock due to Improper Grounding

• To prevent an electric shock hazard, provide proper

grounding connection.
(Sections 1.7)

Electric Shock due to Contact with Inside of Instrument

• If you touch the internal electric parts of the instrument, you

may receive an electric shock. To prevent this, refer
inspection of the internal parts to qualified service personnel.
(Sections 6.6)

SAFETY - 5
 SAFETY SUMMARY
CAUTION Indications

Injury due to Gazing Directly at Xe Flash Lamp When Lit

• The Xe flash lamp radiates strong ultraviolet rays. Avoid

gazing at it directly when lit. For looking at the lamp, be sure
to wear goggles equipped with a function for cutting off
ultraviolet rays.
(Sections 5.2)

Fatigue due to Long-Hour Operation

• If you keep working with the display monitor and keyboard for
long hours, your eyes and body will be fatigued to jeopardize
your health. When working with the display monitor for a
long time, take a break for 10 to 15 minutes per hour for the
health of your eyes and body.
(Chapter 3)

Heavy Instrument

• This instrument weight as much as 15 kg. When carrying

this instrument, exercise care not to incur injury by dropping it
accidentally. Be sure to hold the right and left parts of the
instrument securely when moving it.
(Sections 1.1)

SAFETY - 6
 SAFETY SUMMARY
NOTICE

Disposal of Waste Solution

Be sure to collect waste solution and treat it for proper disposal

in accordance with the relevant laws and regulations regarding
water pollution control and sewage treatment.
Improper treatment of waste solution may result in environmental
pollution, and could also lead to a penalty.

Accuracy and Precision of Measured Values

Carry out periodic inspection and check whether the system is

operating normally. If necessary, conduct measurement on a
control sample.

Burst of a Lithium Battery

This spectrophotometer uses a lithium battery to backup the data

in the memory. A lithium battery may burst should it be handled
improperly. Be sure not to charge and dissolve it, or throw it to
the fire under any circumstance. It should be handled totally
separate from ordinary wastes.
When the lithium battery needs to be replaced (for example,
when an error message RAM NG is displayed frequently on the
screen), contact the dealer from whom you bought the
spectrophotometer or the nearest service department.
Please entrust the replacement to a service agent who finished
the technical training provided by our company.
(Replacement is charged after the warranty period of the
spectrophotometer.

SAFETY - 7
 SAFETY SUMMARY
NOTICE (Continued)

Electrical Checkups

• Make sure that power supply to the spectrophotometer is

100 V AC, 1 kVA or more (50 or 60 Hz). Voltage fluctuation
or noise entering the power line may have an adverse effect
on the spectrophotometer main unit and cause a malfunction
as well.

• Be sure to prepare a grounding wire together with power

cables and confirm that the wire has a ground resistance of
100 Ω or less (Class D grounding construction in Electric
Facility Technical Standards). Insufficient grounding may
make the instrument susceptible to noise from the exterior
and cause a stray voltage to appear in the main unit, which
would be hazardous to personnel.

SAFETY - 8
 CONTENTS

PREFACE

ABOUT THIS MANUAL

IMPORTANT ................................................................................................IMPORTANT-1
Warranty on Product ........................................IMPORTANT-1
Installation, Relocation and After-sale
Technical Service.............................................IMPORTANT-4
Technical Seminars and Training Courses
for Users...........................................................IMPORTANT-5
Caution on Disposal of Instrument ...................IMPORTANT-5
Service Life of This Instrument.........................IMPORTANT-6
Other Precautions ............................................IMPORTANT-6

SAFETY SUMMARY ....................................................................................SAFETY-1

General Safety Guidelines ..................................SAFETY-1
Common Safety Precautions ..............................SAFETY-2
Prior to Use .........................................................SAFETY-2
In Use..................................................................SAFETY-3
Installation, Maintenance, and Relocation ..........SAFETY-3
Safety Instructions in This Manual ......................SAFETY-5
DANGER Indications ..........................................SAFETY-5
WARNING Indications ........................................SAFETY-5
Electric Shock due to Contact with
Dangerous High Voltage (100 V) ...................SAFETY-5
Electric Shock due to Improper Grounding ....SAFETY-5
Electric Shock due to Contact with Inside
of Instrument ..................................................SAFETY-5
CAUTION Indications..........................................SAFETY-6
Injury due to Gazing Directly at Xe Flash
Lamp When Lit ...............................................SAFETY-6
Fatigue due to Long-Hour Operation..............SAFETY-6
Heavy Instrument ...........................................SAFETY-6
NOTICE ..............................................................SAFETY-7
Disposal of Waste Solution ............................SAFETY-7
Accuracy and Precision of Measured
Values ............................................................SAFETY-7
Burst of a Lithium Battery ...............................SAFETY-7
Electrical Checkups........................................SAFETY-8

-i-
1. INSTALLATION (REFERENCE INFORMATION FOR USER) ............................... 1-1
1.1 Unpacking ............................................................... 1-1
1.2 Installation Conditions ............................................. 1-2
1.2.1 Power Supply.............................................. 1-2
1.2.2 Installation Space........................................ 1-2
1.3 Environmental Conditions ....................................... 1-3
1.3.1 Operating Temperature............................... 1-3
1.3.2 Operating Humidity ..................................... 1-3
1.3.3 Storage Temperature.................................. 1-3
1.3.4 Storage Humidity ........................................ 1-3
1.3.5 Atmospheric Gas ........................................ 1-3
1.3.6 Other General Precautions ......................... 1-4
1.4 Check of Contents................................................... 1-5
1.5 Check of Power Voltage and Fuse .......................... 1-5
1.6 Connection of Cords................................................ 1-6
1.7 Connection of Power Cord and Ground Wire .......... 1-7

2. FUNCTIONS AND BASIC OPERATION ................................................................ 2-1

2.1 Features of Model U-5100....................................... 2-1
2.2 Name and Function of Each Part ............................ 2-1
2.2.1 Names and Functions of Parts of
Spectrophotometer Main Unit ..................... 2-1
2.2.2 Inside View of Sample Compartment.......... 2-4
2.3 Startup and Shutdown of Instrument ....................... 2-6
2.3.1 Startup of Instrument .................................. 2-6
2.3.2 Shutdown of Instrument .............................. 2-8
2.4 Basic Operation....................................................... 2-9
2.4.1 Operation Panel .......................................... 2-9
2.4.2 Basic Operation of Screen .......................... 2-12
2.4.3 How to Input Characters ............................. 2-15
2.4.4 How to Set Cell ........................................... 2-17
2.4.5 Demounting and Remounting of
6-cell Holder................................................ 2-19
2.4.6 Cautions on Operation ................................ 2-22
2.5 Basic Setting ........................................................... 2-23
2.5.1 Setting of Current Date and Time ............... 2-23
2.5.2 Setting of Printer ......................................... 2-25
2.5.3 Setting of Auto Lamp OFF Time ................. 2-27
2.5.4 Setting of 6-cell Mode ................................. 2-29
2.5.5 Selection of Language ................................ 2-31

3. FIRST MEASUREMENT ......................................................................................... 3-1

3.1 What this Product can Do........................................ 3-1
3.2 Automatic Continuous Measurement (6-cell
auto mode) .............................................................. 3-2

- ii -
 3.2.1 Determining Concentration of Solution ....... 3-3
3.2.2 Measuring Absorbance/Transmittance ....... 3-41
3.2.3 Measuring DNA Sample (measurement
through ratio calculation)............................. 3-60
3.2.4 Measuring a Spectrum................................ 3-79
3.3 Sample-by-sample Measurement (6-cell manual
mode) ...................................................................... 3-97
3.3.1 Determining Concentration of Solution ....... 3-98
3.3.2 Measuring Absorbance/Transmittance ....... 3-114
3.3.3 Measuring DNA Sample (measurement
through ratio calculation)............................. 3-121
3.3.4 Measuring a Spectrum................................ 3-128
3.3.5 Measuring Change with Time ..................... 3-134
3.4 Measurement with Enlarged Display Screen
(monitor) .................................................................. 3-147

4. FOR MORE EFFICIENT OPERATION ................................................................... 4-1

4.1 Loading or Deleting the Saved Data ....................... 4-1
4.1.1 Loading the Saved Data ............................. 4-1
4.1.2 Deleting the Saved Data ............................. 4-4
4.1.3 Printing the List of Saved Data ................... 4-7
4.1.4 Photometry Based on Saved Calibration
Curve Data.................................................. 4-10
4.2 Loading and Deleting the Saved Method ................ 4-14
4.2.1 Measurement via Loading of Saved
Method ........................................................ 4-14
4.2.2 Deletion of Saved Method........................... 4-17
4.2.3 Printing the List of Saved Methods ............. 4-20
4.3 Data Processing ...................................................... 4-23
4.3.1 Scale Change of Calibration Curve............. 4-23
4.3.2 How to Check Calibration Curve Factor,
Correlation Coefficient and Determination
Coefficient ................................................... 4-26
4.3.3 Deletion and Recovery of Calibration
Curve Data.................................................. 4-29
4.3.4 Scale Change (Wavelength/Time Scan
Data) ........................................................... 4-31
4.3.5 Peak Detection (Wavelength Scan
Data) ........................................................... 4-35
4.3.6 Data Tracing (Wavelength/Time Scan
Data) ........................................................... 4-38
4.3.7 Data Smoothing (Wavelength/Time Scan
Data) ........................................................... 4-41
4.3.8 Displaying Data in List (Wavelength/Time
Scan Data) .................................................. 4-44

- iii -
 4.4 Measurement in Statistical Calculation Mode ......... 4-47
4.5 Auto Start Function.................................................. 4-49
4.6 Introduction and Mounting Method of Separately
Available Options .................................................... 4-52
4.6.1 Single Cell Holder (Separately Available) ... 4-53
4.6.2 Mask for Micro Cell (Separately
Available) .................................................... 4-55
4.6.3 Rectangular Long-Path Cell Holder
(Separately Available)................................. 4-57
4.6.4 How to Return 6-Cell Holder ....................... 4-59
4.6.5 Measurement in 6-Cell Mode OFF
Status.......................................................... 4-60

5. PERFORMANCE CHECK....................................................................................... 5-1

5.1 Check with Instrument Main Unit Alone .................. 5-1
5.1.1 Wavelength Accuracy ................................. 5-3
5.1.2 Wavelength Repeatability ........................... 5-7
5.1.3 Noise Level (RMS)...................................... 5-10
5.1.4 Baseline Flatness........................................ 5-13
5.1.5 Baseline Stability......................................... 5-16
5.1.6 Hardware Check ......................................... 5-19
5.1.7 Report Printing ............................................ 5-21
5.1.8 Auto Check ................................................. 5-22
5.2 Check with Pen Type Low-Pressure Mercury
Lamp Available at Option ........................................ 5-24
5.2.1 Wavelength Accuracy (with Hg Lamp)........ 5-26
5.2.2 Wavelength Repeatability (with Hg
Lamp).......................................................... 5-28
5.2.3 Resolution ................................................... 5-30
5.2.4 Report Printing ............................................ 5-32
5.3 Wavelength Calibration ........................................... 5-33
5.3.1 Wavelength Calibration Method.................. 5-33

6. MAINTENANCE ...................................................................................................... 6-1

6.1 Lamp Usage ............................................................ 6-1
6.2 How to Clean the Instrument ................................... 6-3
6.3 Cleaning and Storing a Sample Cell ....................... 6-4
6.4 Replacement of Radiation Source Lamp................. 6-4
6.5 Caution on Lithium Battery ...................................... 6-4
6.6 Replacement of Power Fuse ................................... 6-5
6.7 Storage of Instrument.............................................. 6-7
6.8 If You Encounter a Trouble ..................................... 6-8
6.9 Specifications of Model U-5010 Ratio Beam
Spectrophotometer.................................................. 6-15

- iv -
APPENDIX ..................................................................................................... APPENDIX-1
Appendix a. Operating Principles of Model
U-5100......................................... APPENDIX-1
Appendix b. Absorption Spectrophotometry.... APPENDIX-4
Appendix c. Proper Use of
Spectrophotometer...................... APPENDIX-5

-v-
 1.1

1. INSTALLATION
(REFERENCE INFORMATION FOR USER)

This instrument shall be installed by service personnel qualified

by Hitachi-Technologies Corporation. The following description
is included as reference information on instrument configuration,
etc. for the user.

1.1 Unpacking

Unpack the shipping crate, carefully take out the

spectrophotometer and place it on a sturdy table or desk.
Take utmost care when taking out the instrument.

CAUTION
Caution on Carrying Heavy Instrument
This instrument weighs about 13 kg. There is a danger of
personal injury if you accidentally drop it. When carrying
the instrument, carefully handle it and securely hold it at
both sides.

Fig. 1-1 Model U-5100 Ratio Beam Spectrophotometer

1-1
1.2 Installation Conditions

1.2 Installation Conditions

Before installation, check the following conditions.

1.2.1 Power Supply

Power voltage : 100, 115, 220, 230 or 240 V

Voltage variation shall be within ±10% of the
rated voltage.
Frequency : 50 or 60 Hz
Frequency variation shall be within ±0.5 Hz of
the rated frequency.
Power capacity : 100 VA or more
Ground line : Grounding resistance 100 Ω or less.

1.2.2 Installation Space

Floor area: 400 mm (W) × 470 mm (D) or more

Provide a space of at least 200 mm at the front and
rear of the main unit.
(Select a level installation position capable of
withstanding a load of 30 kg and at a height of
300 mm or more above floor level.)

1-2
 1.1

1.3 Environmental Conditions

1.3.1 Operating Temperature

15 to 35 °C
For measurement under the most stable conditions, it is
recommended to install the instrument in a room which is air-
conditioned at 20 to 25 °C.

1.3.2 Operating Humidity

25 to 80%
No condensation is allowed.
At 30 °C or higher, keep a humidity of 70% or less.

1.3.3 Atmospheric Gas

(1) Acidic, alkaline and other gases which may corrode metals
significantly shall not be contained in the atmosphere.

(2) Gaseous organic solvents (particularly benzene and

thinner) which may dissolve paint.

1.3.4 Other General Precautions

(1) The instrument should not be exposed to direct sunlight

(which may deteriorate the optical performance or discolor
the housing. Avoid a position near a window.)

(2) Sensible vibration or shock should not be applied to the

instrument (otherwise the fine adjustment or mechanism
might get out of order).

(3) There should be no heat source such as a gas burner,

electric heater and oven near the instrument to prevent the
main unit cover being heated (to 70 °C or higher).

(4) The instrument should not be located near a device

producing a strong electric field (electric welding machine,
high-frequency electric furnace or pole transformer for
example).

1-3
1.3 Environmental Conditions

(5) The installation site should be free from excessive dust

(which may deteriorate optical performance).

(6) The instrument should be free from abrupt variation in

power voltage (which constitutes a noise source).

(7) Avoid frequently turning on/off the electric motor not

provided with a noise suppressor (of a stirrer, vibrator, etc.)
on the power line connected with the spectrophotometer
main unit.

NOTICE: The optical system in the spectrophotometer is

very delicate. And the control unit incorporates
parts constructing a high-density electronic
circuit which functions as a computer.
So careful consideration should be given to the
above items.

1-4
 1.4

1.4 Check of Contents

After unpacking, check the contents against the packing list.

If any part is missing or damaged, or if you have a question,
contact the dealer from whom you purchased the instrument.

1.5 Check of Power Voltage and Fuse

WARNING
Electric Shock due to Dangerous Voltage
An electric shock due to power voltage could result in death
or serious injury. Before connecting the power cord, make
sure that the power switch of the spectrophotometer main
unit is turned off.
Also, absolutely avoid disassembling or modifying the
instrument.

Check that the power line voltage to be applied meets the

operating voltage requirement of the spectrophotometer which is
indicated near its power connector.

Fuse holder

Fig. 1-2 Power Connector Section at Instrument Rear

Referring to section 6.6, check if a fuse having the capacity

indicated in Table 1-1 is used.

Table 1-1 Fuse Capacity and Part No.

Fuse Capacity Part No.

2 A (time lag) J821391

1-5
1.6 Connection of Cords

1.6 Connection of Cords

Connect the cords referring to Fig. 1-3.

WARNING
Electric Shock due to Dangerous Voltage
An electric shock due to power voltage could result in death
or serious injury. Before connecting the power cord, make
sure that the power switch of the spectrophotometer main
unit is turned off.

To AC power supply

Power cable

Printer cable

Printer

Printer
Model U-5100
Spectrophotometer

Fig. 1-3 Connection of Cords (for independent operation of

instrument)

NOTE: Separately prepare a printer having <1> and <2> given

below.
<1> Centronics cable
<2> Any of the following printer languages
(commands)

Printer Language
(command)
DPU
ESC/P V.2
PCL3

1-6
 1.7

1.7 Connection of Power Cord and Ground Wire

(1) Securely plug the power cord into the connector of the main
unit.

(2) When using a plug adapter or table tap, securely connect

its ground wire to the ground terminal.

WARNING
Improper grounding may result in an electric shock hazard.
Be sure to provide proper grounding connection.

To power outlet

Ground
Grounding terminal
resistance:
100 Ω or less Spectrophotometer

Ground wire (green)

Power cord
Underground 75 cm or more E910014 or E910001
underground
Copper plate

300 mm

Class D grounding
installation of electric
facility technical
standards)
2 mm

10 mm

COMMON HOT
COMMON S
W White

GROUND

HOT HOT GROUND COMMON

S Black Green
GROUND

Fig. 1-4 Grounding Connection

1-7
 2.2.1

2. FUNCTIONS AND BASIC OPERATION

2.1 Features of Model U-5100

This instrument is capable of quantitatively determining a liquid

sample and measuring the absorbance, transmittance,
absorption spectrum, transmission spectrum and
absorbance/transmittance change with time.
With this instrument, the light source is turned on only during
measurement. The instrument can thus save a larger amount
of electric power than the spectrophotometer with a continuous
light source. Also, the instrument is standard-equipped with the
automatically turning 6-cell measurement function, thus
permitting quantitative analysis through simple operation.

2.2 Name and Function of Each Part

2.2.1 Names and Functions of Parts of Spectrophotometer Main Unit

Data display

Sample
compartment

Operation
panel

Power switch
(lit in blue when
turned on)

Fig. 2-1 External View of Model U-5100 Ratio Beam Spectrophotometer

2-1
2.2 Name and Function of Each Part

<1> Operation panel : By using the keys on this panel, you

can operate the spectrophotometer.
<2> Data display : While watching this display panel,
operate the spectrophotometer and
check data.
<3> Sample
compartment : In this compartment, set a sample to be
measured.
<4> Power switch : Used to turn on/off the power supply.

<5> Remote selector switch

<6> Printer output connector

<7> LCD brightness control

Fig. 2-2 Left Side View of Spectrophotometer

<5> LOCAL/REMOTE
selector switch : Set this switch to the LOCAL side.
<6> Printer output
connector : Connect the printer cable to this
connector.
<7> LCD brightness
control : Used to adjust the LCD brightness.

2-2
 2.2.1

<8> Light source cover

Fig. 2-3 Rear View of Spectrophotometer

<8> Light source

cover : Opened/closed only when mounting
the pen-shaped low-pressure mercury
lamp (optionally available). For how
to mount the pen-shaped low-pressure
mercury lamp, refer to its instruction
manual.

<9> Drain

Fig. 2-4 Bottom View of Spectrophotometer

2-3
2.2 Name and Function of Each Part

<9> Drain : Tube for draining liquid accidentally

spilt in the sample compartment.
The spilt liquid is discharged to the
bottom of the instrument through this
drain. If you accidentally spill liquid in
the sample compartment, then carry
out cleaning according to section 6.2.

2.2.2 Inside View of Sample Compartment

An inside view of the sample compartment plus the 6-cell holder

and drain are described below.

6-cell holder

Drain

Fig. 2-5 Inside of Sample Compartment

Designation of cell
position

Fig. 2-6 Top View of 6-cell Holder

2-4
 2.2.2

6-cell holder:
Set cells in the 6-cell holder for measurement. This cell holder
can accommodate 6 cells. The cell position inscribed with
Autozero is used only for auto zero. Normally, auto zero is
executed with the cell at this position (hereafter referred to as
cell A). The cell positions inscribed with 1 to 5 are used for
sample measurement (hereafter referred to as cell 1, cell 2, ...
and cell 5).

Drain:
Tube for draining liquid accidentally spilt in the sample
compartment. The spilt liquid is discharged to the bottom of the
instrument through this drain. If you accidentally spill liquid in
the sample compartment, then carry out cleaning according to
section 6.2.

2-5
2.3 Startup and Shutdown of Instrument

2.3 Startup and Shutdown of Instrument

2.3.1 Startup of Instrument

1. Power on the Instrument.

(1) Check that the power switch lamp on the front of the
instrument is extinguished.

(2) Check that the LOCAL/REMOTE selector switch on the

left side of the instrument is set at the LOCAL side.

(3) Check that the power cable is connected between the

instrument and power outlet.

(4) Check that no sample is set in the sample

compartment. If any sample is set, take it out of the
sample compartment.

(5) Close the cover of the sample compartment.

(6) Press the power switch on the front of the instrument.

2. Display of Initialization Screen

(1) The initialization screen (Fig. 2-7) appears on the

display panel of the instrument.

Initializing...
HITACHI U-5100 Spectrophotometer

P/N : 3J25300-00
Copyright (C) Hitachi
High-Technologies Corporation 2010

Serial No. 1234-123

ROM Check OK
RAM Check OK
Lamp ON OK Usage: 1%
WL Initialize OK
WL Check OK

Fig. 2-7 Initialization Frame

2-6
 2.3.1

(2) Self-diagnosis and automatic adjustment are carried

out. The items to be checked are listed below.
For each item, “OK” is indicated if its function is normal
and “NG” if abnormal. If “NG” is indicated for ROM
Check, RAM Check, Lamp ON, WL Initialize or WL
Check, the system will stop. In this case, refer to “b.
Troubleshooting” in section 6.8. If “NG” is indicated
for RAM Check or Lamp ON, the system control can be
advanced by pressing the [CLEAR] key.
Upon normal completion of initialization, the Main
Menu screen will appear.

• ROM Check : ROM check

• RAM Check : RAM check
• Lamp ON : The system checks if the lamp is
turned on.
• Lamp Usage : For the lamp, the status of use is
indicated in percentage. A value
of 100% is a standard for service
life. If 100% is exceeded, then
execute performance check with
reference to section 5.1 and make
sure that the performance is within
the scope of the specifications.
If outside the scope of
specifications, contact the dealer
from whom you purchased the
instrument or your nearest
maintenance service office.
• WL Initialize : Check of wavelength drive system
• WL Check : Wavelength calibration at 484.6 nm

3. Display of Main Menu Screen

The Main Menu screen (Fig. 2-8) appears. From this

screen, you can proceed to each measurement item or
instrument setup item.
After starting the instrument, 2-hour warming-up is
necessary for the measurement requiring an instrument
stability equivalent to the specified baseline stability level
(0.0007 Abs/h) (for time scan or measurement in which
auto zero is not executed for a long time).

2-7
2.3 Startup and Shutdown of Instrument

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 2-8 Main Menu Screen

2.3.2 Shutdown of Instrument

1. Termination of Measurement

(1) Check that the power switch lamp on the front of the
instrument is lit.

(2) When measurement is in progress, wait until it is

terminated or press the [STOP] key. When monitor
display is provided, press the [RETURN] key.

(3) Save or print out data if desired. For details of save

and print operations, refer to Section 3.

2. Power off the Instrument.

Press the power switch on the front of the instrument to turn

off power. Check that the power switch lamp is
extinguished.

NOTICE: Avoid turning off the power switch immediately

after performing data save operation.
Otherwise data might not be saved. Wait for at
least 5 seconds or so after the save operation.

2-8
 2.4.1

2.4 Basic Operation

2.4.1 Operation Panel

Figure 2-9 shows the operation panel of the instrument.

The function of each key is explained in Table 2-1, and key
operation in combination with the [SHIFT] key in Table 2-2.

<5> <6> <7>

<1>

<2> <8>

<3> <13>
<10>
<4>

<9> <11> <12>

Fig. 2-9 Operation Panel

Table 2-1 Independent Key Operation (The bold-faced characters are entered in
sequence each time the relevant key is pressed.)

Key Description
<1> MAIN MENU Displays the Main Menu screen.
<2> MEAS.SCREEN Determines the current measuring conditions and changes the currently
displayed screen to the measurement screen.
<3> AUTO ZERO Adjusts the photometric value to 0 Abs in ABS mode and to 100%T in
%T mode.
<4> SHIFT By holding down this key and pressing another one, you can execute a
different command.
<5> 1 Enters 1.
On the character input screen, the following characters are entered.
Alphabet input : +, _, /, :, *, (, ), ;, 1
Numeric input : 1
2 Enters 2.
On the character input screen, the following characters are entered.
Alphabet input : a, b, c, A, B, C, 2
Numeric input : 2
3 Enters 3.
On the character input screen, the following characters are entered.
Alphabet input : d, e, f, D, E, F, 3
Numeric input : 3

2-9
2.4 Basic Operation

(cont’d)
Key Description
<5> 4 Enters 4.
On the character input screen, the following characters are entered.
Alphabet input : g, h, i, G, H, I, 4
Numeric input : 4
5 Enters 5.
On the character input screen, the following characters are entered.
Alphabet input : j, k, l, J, K, L, 5
Numeric input : 5
6 Enters 6.
On the character input screen, the following characters are entered.
Alphabet input : m, n, o, M, N, O, 6
Numeric input : 6
7 Enters 7.
On the character input screen, the following characters are entered.
Alphabet input : p, q, r, s, P, Q, R, S, 7
Numeric input : 7
8 Enters 8.
On the character input screen, the following characters are entered.
Alphabet input: t, u, v, T, U, V, 8
Numeric input: 8
9 Enters 9.
On the character input screen, the following characters are entered.
Alphabet input : w, x, y, z, W, X, Y, Z, 9
Numeric input : 9
⎯ Enters “-”(hyphen).
0 Enters 0.
. Enters “.” (period).
<6> GO TO WL Used for shifting the wavelength. Upon pressing this key, the guidance
given below appears. You can shift the wavelength by entering a
wavelength in a range of 190.0 to 1100.0 (in steps of 0.1 nm).
Input value. :_
WL(nm) [190.0 - 1100.0nm]

<7> PRINT Used for printing out data. Pressing this key after measurement prints
out a report. When it is pressed with the monitor screen displayed, the
current wavelength and photometric value are printed out. If pressed
with the data read by the cursor displayed, the cursor data is printed out.
<8> ► Used for moving the cursor to the right on the Meas. PARAM. setting
screen. Also used for moving to the next page when ◄ ► is indicated
in the lower part of the screen.
◄ Used for moving the cursor to the left on the Meas. PARAM. setting
screen. Also used for moving to the previous page when ◄ ► is
indicated in the lower part of the screen.
▲ Used for moving up the cursor.
▼ Used for moving down the cursor.
<9> ENTER Used for determining the entered character/numerical value or the set
item.
<10> CLEAR Used for erasing the entered character or numerical value.
<11> RETURN Used for returning to the previous screen.
<12> START Used for starting measurement, proceeding to the next measurement or
advancing to the next measurement screen.
<13> STOP Used for stopping measurement.

2 - 10
 2.4.1

Table 2-2 Key Operation in Combination with SHIFT Key (The bold-faced characters
are entered in sequence each time the relevant key is pressed.)

Key Description
[SHIFT] + Allows you to change the currently displayed screen to the monitor screen.
MEAS.SCREE
N
[SHIFT] + 1 On the character input screen, the following characters are entered.
Alphabet input : 1
Numeric input : +, _, /, :, *, (, ), ;, 1
[SHIFT] + 2 On the character input screen, the following characters are entered.
Alphabet input : 2
Numeric input : a, b, c, A, B, C, 2
[SHIFT] + 3 On the character input screen, the following characters are entered.
Alphabet input : 3
Numeric input : d, e, f, D, E, F, 3
[SHIFT] + 4 On the character input screen, the following characters are entered.
Alphabet input : 4
Numeric input : g, h, i, G, H, I, 4
[SHIFT] + 5 On the character input screen, the following characters are entered.
Alphabet input : 5
Numeric input : j, k, l, J, K, L, 5
[SHIFT] + 6 On the character input screen, the following characters are entered.
Alphabet input : 6
Numeric input : m, n, o, M, N, O, 6
[SHIFT] + 7 On the character input screen, the following characters are entered.
Alphabet input : 7
Numeric input : p, q, r, s, P, Q, R, S, 7
[SHIFT] + 8 On the character input screen, the following characters are entered.
Alphabet input : 8
Numeric input : t, u, v, T, U, V, 8
[SHIFT] + 9 On the character input screen, the following characters are entered.
Alphabet input : 9
Numeric input : w, x, y, z, W, X, Y, Z, 9
[SHIFT] + - Displays the data loading screen.
[SHIFT] + 0 Displays the method loading screen.
[SHIFT] + . Displays the 6-cell move screen. The guidance given below will appear.
By entering a cell number, it is possible to move to the target cell position directly.
Input value. :_
Cell No.[0 - 5, 0:for Autozero]

[SHIFT] + Changes over between alphabet input and numeric input on the character input
GO TO WL screen.
[SHIFT] + Feeds the printer paper. Note that this key operation is invalid when the printer
PRINT for PCL3 commands is used.
[SHIFT] + ► Turns the 6-cell holder counterclockwise.
[SHIFT] + ◄ Turns the 6-cell holder clockwise.
[SHIFT] + Clears a character before the cursor to move it back one position.
CLEAR

2 - 11
2.4 Basic Operation

2.4.2 Basic Operation of Screen

Explained here is the gateway to operation or the Main Menu

screen. For the menu items and setting items displayed on the
monitor, you can proceed with operation by use of the [◄], [►],
[▲] and [▼] keys, numeric keys and [ENTER] key on the
operation panel.

a. Main Menu Screen and Basic Operation

Start the instrument referring to section 2.3.1. The Main

Menu screen will appear.

<1> Screen title

Main Menu CELL A

1 Photometry
2 WL Scan <5> Guidance on
3 Time Scan cell position
4 Method Menu
5 Data Menu
6 System Setup <3> Cursor
7 Performance <2> Menu
8 Data Display

Select function. : _

<4> Operational guidance

<1> Screen title : Title of the currently displayed screen.

<2> Menu : Selectable menu items are indicated.
<3> Cursor : Indicates the currently selected item.
The cursor can be moved up and down
by pressing the [▲] and [▼] keys.
Move the cursor to the aimed-at item
and press the [ENTER] key, and you
can access to its details.

2 - 12
 2.4.2

<4> Operational
guidance : Guidance on operation is given for the
menu item selected by the cursor.
This guidance differs depending on the
screen active at the time. In the
subsequent process, carry out
operation according to the guidance.
On the Main Menu screen, “Select
function.: _” is indicated. By entering
the number preceding each menu item,
you can proceed to internal setting.
<5> Guidance on
cell position : Indicates the current measurement
position when the 6-cell holder is
mounted. If any other accessory than
the 6-cell holder, such as a single cell
holder, rectangular long path cell
holder or auto sipper (optionally
available), is used, indication varies
with the accessory. For details, refer
to Tables 2-3 and 2-4.

Table 2-3 Guidance on Cell Position in Use of 6-cell Holder

Guidance on
Status of 6-cell Holder
Cell Position
CELL A Indicated when “Autozero” on the 6-cell holder is at
the measurement position.
CELL 1 Indicated when “1” on the 6-cell holder is at the
measurement position.
CELL 2 Indicated when “2” on the 6-cell holder is at the
measurement position.
CELL 3 Indicated when “3” on the 6-cell holder is at the
measurement position.
CELL 4 Indicated when “4” on the 6-cell holder is at the
measurement position.
CELL 5 Indicated when “5” on the 6-cell holder is at the
measurement position.
CELL * Indicated when the 6-cell holder is turning.
CELL - Indicated when the cell position on the 6-cell holder is
not yet pinpointed.

2 - 13
2.4 Basic Operation

Table 2-4 Guidance on Cell Position in Use of Accessory

Guidance on
In-use Accessory Indicated
Cell Position
CELL S Single cell holder (optionally available)
Rectangular long path cell holder (optionally
available)
SIPPER Auto sipper (optionally available)

To display the Main Menu screen from another screen,

press the [MAIN MENU] key.

b. Measuring Condition Setting Screen

Described below is the basic operation of the measuring

condition setting screen when “Photometry” has been
selected on the Main Menu screen.

Photometry CELL A
1 Meas.PARAM. Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function.:_

Fig. 2-10 Measuring Condition Setting Screen

Two methods are available for the setting of measuring

conditions. Select either of them from the perspective of
your ease of use, or use them in combination.

1. Use of [◄], [►], [▲] and [▼] keys

Each item on the screen can be selected with the [◄],
[►], [▲] and [▼] keys. For setting “3 Curve” for
example, move the cursor to “3 Curve” with the [▲] or
[▼] keys and press the [►] key to enable setting of
each item in “3 Curve”. Set the conditions according
to the guidance. The cursor can be moved to the left
with the [◄] key.

2 - 14
 2.4.3

2. Use of numeric keys and [ENTER] key

Enter the number preceding the desired menu item
and press the [ENTER] key. For setting “3 Curve” for
example, enter <3> and press the [ENTER] key.
Then, set each item according to the operational
guidance and press the [ENTER]. To move the
cursor to the left, press the [RETURN] key.

2.4.3 How to Input Characters

Explained below is how to input characters. Character input is

done when entering the file name, sample name and
concentration unit.

a. Character Input Screen

Character input is done when entering the file name,

sample name and concentration unit. On each input
screen, the guidance shown in Fig. 2-11 appears. For the
file name, you can input up to 20 characters, and up to 8
characters for the sample name and concentration unit.

File Name：
File Name [Max: 20 characters]
ABC W X ( 1/ 1)
Indication of character
File name input screen input mode
ABC : Alphabet input
Input character.:_ 123 : Numeric input
Sample Name[Max: 8 characters]
ABC

Sample name input screen

Input character.:_
Unit Label[Max: 8 characters]
ABC

Concentration unit input screen

Fig. 2-11 Character Input Screen

2 - 15
2.4 Basic Operation

b. Alphabet Input and Numeric Input

Alphabet input mode and numeric input mode are available

for character input. In alphabet input mode, “ABC” is
indicated in the lower part of the screen, and “123” is
indicated in numeric mode. Alphabet input mode is initially
set on the character input screen. To change over
between alphabet input and numeric input, hold down the
[SHIFT] key and press the [GO TO WL] key.
In alphabet input mode, you can enter a numeric character
in numeric mode by holding down the [SHIFT] key and
pressing the numeric (1 to 9) key. In numeric input mode,
you can enter a character in alphabet input mode.
<Example>
When you hold down the [SHIFT] key and press the <5>
key in alphabet input mode, “5” is entered.
When you hold down the [SHIFT] key and press the <5>
key in numeric input mode, “j” is entered.

c. Character Transition in Alphabet Input Mode

In alphabet input mode, the character entered is changed

over as follows by pressing the key repeatedly:
Small letters of alphabet assigned to each key → Capital
letters of alphabet → numeric on each key
<Example>
With the <3> key, the character is repeatedly changed over
in the order of d → e → f → D → E → F → 3. (For the
other keys, refer to Table 2-1.)

d. Other Operations

For erasing one character, press the [CLEAR] key.

By pressing the [►] key, you can enter the same
alphabetical character repeatedly or place a space.
To determine the entered character(s), press the [ENTER]
key.

2 - 16
 2.4.4

2.4.4 How to Set Cell

This section explains about the cell, including cell selection,

sample volume required for measurement and how to set a cell
to the 6-cell holder.

a. About Cell

10 mm cell is generally used for the spectrophotometer.

This cell is 10 mm on an inner side.

Table 2-5 Type of Cell

Cell Name P/N Measurable Sample Volume

10 mm quartz cell 123-1004 1.7 to 3.5 mL
(optionally available)
10 mm glass cell 123-1010 1.7 to 3.5 mL
(optionally available)

b. Setting of Cell

The cell has transparent and frosted faces. It should be

held by the frosted faces. If held by the transparent faces,
its measurement face will be contaminated with fingerprints,
thus causing an error in measurement.

Frosted face Frosted face

(Hold by this face.) (Hold by this face.)

Transparent face
(measurement face)

Fig. 2-12 10 mm Cell Holder

2 - 17
2.4 Basic Operation

For measurement, set the cell so that the beam passes

through its transparent face. Figure 2-13 shows the
external view of the 6-cell holder. It turns for measurement
of each cell. It is therefore necessary to set the cell in
different directions according to the cell position.
Triangular marks are put on the 6-cell holder to indicate the
incident direction of beam. So set each cell so that its
transparent face is matched with the triangular mark.

The beam passes through

the face matched with the
triangular mark.

Fig. 2-13 External View of 6-cell Holder

c. Other Cells

In addition to the above-mentioned cell, you can use a

micro cell (sample volume: 340 to 600 µL) by utilizing the
optionally available single cell holder and micro cell mask.
You can also use a cell whose optical path length is
100 mm (sample volume: 17 to 35 mL) by utilizing the
optionally available rectangular long path cell holder.
For details of these options, refer to section 4.6.

2 - 18
 2.4.5

2.4.5 Demounting and Remounting of 6-cell Holder

The 6-cell holder is demounted when installing any accessory or

cleaning the sample compartment. Described below are the
procedures for demounting and remounting the holder.

a. Procedure for Demounting 6-cell Holder

(1) Loosen the hand screw at the center of the 6-cell

holder until being turned freely.

Hand screw

6-cell holder

Fig. 2-14 Preparation for Demounting of

6-cell Holder

(2) Hold the 6-cell holder by the top and vertically pull it
out of the sample compartment.

6-cell holder

Fig. 2-15 Demounting of 6-cell Holder

2 - 19
2.4 Basic Operation

(3) Put the demounted 6-cell holder in a case which can

protects against dust and keep it in a safe place.

6-cell holder

Fig. 2-16 Demounted 6-cell Holder

b. Procedure for Remounting 6-cell Holder

Given below is the procedure for remounting the 6-cell

holder demounted.

(1) Open the sample compartment. Check the 6-cell

holder driving shaft in the sample compartment.
The shaft has a notched portion (concave portion).
Let us refer to this portion as A.

Drive shaft in sample

compartment

Notched
portion (A)

Top view of
drive shaft

Fig. 2-17 Drive Shaft in Sample Compartment

2 - 20
 2.4.5

(2) Check the screw cramp section on the back side of the
6-cell holder to be remounted. The screw cramp
section includes a portion for alignment (convex
portion). Let us refer to this portion as B. Align the
6-cell holder by turning it so that the convex portion for
alignment fits into the notched portion of the drive shaft
in the sample compartment. Then, fit the 6-cell holder
to the drive shaft.

Screw cramp Portion for

section alignment (B)

Screw

Screw clamp section on

back side of 6-cell holder

Fig. 2-18 Back Side View of 6-cell Holder

(3) Fix the 6-cell holder by tightening the hand screw at

the center. The 6-cell holder has now been
remounted.

Hand screw

Fig. 2-19 Fixing of 6-cell Holder

2 - 21
2.4 Basic Operation

2.4.6 Cautions on Operation

Pay attention to the following points during operation and

measurement.

NOTICE: For sample measurement, securely close the

sample compartment cover. Also, avoid
opening the sample compartment cover during
measurement. Otherwise abnormal measured
values may be obtained.

NOTICE: The main unit has a function of saving the

measured data or measuring conditions.
However, the saved data may be erased if the
lithium battery for memory backup dies or
deteriorates. It is recommended to keep the
backup copy of valuable data on paper, etc.
without fail.

2 - 22
 2.5.1

2.5 Basic Setting

2.5.1 Setting of Current Date and Time

Set the current date and time. The date and time set here are
used for the date and time of analysis, printing, etc.

1. Startup of Instrument

Start the instrument according to section 2.3.1.

2. Main Menu Screen

The Main Menu screen (Fig. 2-20) appears. To select

“System Setup”, press the <6> key (System Setup) and
then [ENTER] key. Or select “System Setup” with the [▲]
or [▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 2-20 Main Menu Screen

3. System Setup Screen

The System Setup screen (Fig. 2-21) appears. To select

“Date and Time”, press the <3> key (Date and Time) and
then [ENTER] key. Or select “Date and Time” with the [▲]
or [▼] key and press the [ENTER] key.

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_

Fig. 2-21 System Setup Screen

2 - 23
2.5 Basic Setting

4. Setting of Date and Time

(1) The Date and Time setting screen (Fig. 2-22) appears.
Select each item indicated and make your entry.
For details, refer to Table 2-6.

System Setup CELL A

1 WL CALIB. Format Y/M/D
2 Lamp Usage Year 2009
3 Date and Time Month 12
4 Printer Setup Day 16
5 6 Cell Mode Hour 22
6 Auto Lamp OFF Minute 7
7 Language

Select function. : _
1 Y/M/D □
□ 2 M/D/Y □
3 D/M/Y

Fig. 2-22 Date and Time Setting Screen

Table 2-6 Date and Time Setting Parameters

Setting Item Description

Format Select a display format for the date.
[1] Y/M/D: Displays the date in year/month/day format.
[2] M/D/Y: Displays the date in month/day/year format.
[3] D/M/Y: Displays the data in day/month/year format.
Year Enter the current year.
Settable in a range of 2008 to 2099.
Month Enter the current month.
Input range: 1 to 12
Day Enter the current day.
Input range: 1 to 31
Hour Enter the current hour.
Input range: 0 to 23
Minute Enter the current minute.
Input range: 0 to 59

(2) After completion of setting, return to the Main Menu

screen with the [RETURN] key.

2 - 24
 2.5.2

2.5.2 Setting of Printer

Select a type of language for the connected printer.

1. Startup of Instrument

Start the instrument according to section 2.3.1.

2. Main Menu Screen

The Main Menu screen (Fig. 2-23) appears. To select

“System Setup”, press the <6> key (System Setup) and
then [ENTER] key. Or select “System Setup” with the [▲]
or [▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 2-23 Main Menu Screen

3. System Setup Screen

The System Setup screen (Fig. 2-24) appears. To select

“Printer Setup”, press the <4> key (Printer Setup) and then
[ENTER] key. Or select “Printer Setup” with the [▲] or [▼]
key and press the [ENTER] key.

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :

Fig. 2-24 System Setup Screen

2 - 25
2.5 Basic Setting

4. Setting of Printer

(1) The printer setup screen (Fig. 2-25) appears. Select

the item indicated and make your entry. For details,
refer to Table 2-7.

System Setup CELL A

1 WL CALIB. Type DPU
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_
1 DPU □
□ 2 PCL3 □
3 ESC/P(Cut)
4 ESC/P(Cont.)
□

Fig. 2-25 Printer Setup Screen

Table 2-7 Printer Setup Parameter

Setting Item Description

Type Select the type of printer language.
[1] DPU : Selected when the printer having
DPU commands is connected.
[2] PCL3 : Selected when the printer having
PCL/3 commands is connected.
[3] ESC/P (Cut) : Selected when connecting the printer
having ESC/P V2 commands and
using cut sheets.
[4] ESC/P (Cont.) : Selected when connecting the printer
having ESC/P V2 commands and
using continuous forms.
If the type of language is unknown, contact the dealer
from whom you purchased the instrument or your nearest
maintenance service office.

(2) After completion of setting, return the Main Menu screen

with the [RETURN] key.

2 - 26
 2.5.3

2.5.3 Setting of Auto Lamp OFF Time

This instrument has a function of automatically turning off the

lamp after a certain period of time to prevent the lamp staying on
unnecessarily for a long time in use of the enlarged display
screen (monitor). You can set this lamp OFF time.
For measurement with the enlarged display screen, refer to
section 3.4.

1. Startup of Instrument

Start the instrument according to section 2.3.1.

2. Main Menu Screen

The Main Menu screen (Fig. 2-26) appears. To select

“System Setup”, press the <6> key (System Setup) and
then [ENTER] key. Or select “System Setup” with the [▲]
or [▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 2-26 Main Menu Screen

3. System Setup Screen

The System Setup screen (Fig. 2-27) appears. To select

“Auto Lamp OFF”, press the <6> key (Auto Lamp OFF) and
then [ENTER] key. Or select “Auto Lamp OFF” with the
[▲] or [▼] key and press the [ENTER] key.

2 - 27
2.5 Basic Setting

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_

Fig. 2-27 System Setup Screen

4. Setting of Auto Lamp OFF Time

(1) The auto lamp OFF time setting screen (Fig. 2-28)
appears. Set the time for automatically turning off the
lamp when the enlarged display screen is active.
For details, refer to Table 2-8.

System Setup CELL A

1 WL CALIB. Lamp OFF 5 min
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_
□1 5 min □
2 15 min □
3 30 min

Fig. 2-28 Auto Lamp OFF Time Setting Screen

Table 2-8 Auto Lamp OFF Setting Parameter

Setting Item Description

Lamp OFF You can set the time for automatically turning
off the lamp when the enlarged display screen
(monitor) is active. It is recommended to
select [1] 5 min unless the enlarged display
screen is frequently used for measurement.
[1] 5 min
[2] 15 min
[3] 30 min

(2) After completion of setting, return to the Main Menu

screen with the [RETURN] key.

2 - 28
 2.5.4

2.5.4 Setting of 6-cell Mode

You can specify whether to control the 6-cell holder.

The setting needs to be changed for measurement with the
optionally available single cell holder or rectangular long path cell
holder in place of the 6-cell holder, or for measurement with the
6-cell holder in place of the single cell holder or rectangular long
path cell holder.

Table 2-9 Setting of 6-cell Mode

Cell Holder 6-cell Mode

6-cell holder ON
Single cell holder OFF
(optionally available)
Rectangular long path cell holder OFF
(optionally available)

1. Startup of Instrument

Start the instrument according to section 2.3.1.

2. Main Menu Screen

The Main Menu screen (Fig. 2-29) appears. To select

“System Setup”, press the <6> key (System Setup) and
then [ENTER] key. Or select “System Setup” with the [▲]
or [▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 2-29 Main Menu Screen

2 - 29
2.5 Basic Setting

3. System Setup Screen

The System Setup screen (Fig. 2-30) appears. To select

“6 Cell Mode”, press the <5> key (6 Cell Mode) and then
[ENTER] key. Or select “6 Cell Mode” with the [▲] or [▼]
key and press the [ENTER] key.

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_

Fig. 2-30 System Setup Screen

4. Setting of 6-cell Mode

(1) The 6-cell mode setting screen (Fig. 2-31) appears.

Select the item indicated and make your entry.
For details, refer to Table 2-10.

System Setup CELL A

1 WL CALIB. Mode ON
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_
1 ON □
□ 2 OFF

Fig. 2-31 6-Cell Mode Setting Screen

Table 2-10 6 Cell Mode Setting Item

Setting Item Description

Mode Select whether to validate control of the 6-cell
holder.
[1] ON : Selected for use of the 6-cell holder.
[2] OFF : Selected for use of the optionally
available single cell holder or rectangular
long path cell holder.

2 - 30
 2.5.5

(2) After completion of setting, return to the Main Menu screen

with the [RETURN] key.

Note：When Method is opened from Method Menu (Refer to

section 4.2.1 for details), Auto Start (Refer to section 4.5 for
details), and data is opened from Data Menu (Refer to section
4.1.1 for details), the setting of ON/OFF of A is subjected to
change.

2.5.5 Selection of Language

You can select a language on the data display between English

and Japanese in the following procedure.

1. Startup of Instrument

Start the instrument according to section 2.3.1.

2. Main Menu Screen

The Main Menu screen (Fig. 2-32) appears. To select

“System Setup”, press the <6> key (System Setup) and
then [ENTER] key. Or select “System Setup” with the [▲]
or [▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 2-32 Main Menu Screen

3. System Setup Screen

The System Setup screen (Fig. 2-33) appears. To select

“Language”, press the <7> key (Language) and then
[ENTER] key. Or select “Language” with the [▲] or [▼]
key and press the [ENTER] key.

2 - 31
2.5 Basic Setting

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_

Fig. 2-33 System Setup Screen

4. Selection of Language

(1) The language selection screen (Fig. 2-34) appears.

Select the item indicated and make your entry.
For details, refer to Table 2-11.

System Setup CELL A

1 WL CALIB. Language English
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. :_
1 English □
□ 2 日本語

Fig. 2-34 Language Selection Screen

Table 2-11 Language Setting Item

Setting Item Description

Language Select a language to be displayed by the instrument.
[1] English : Sets English display.
[2] 日本語 : Sets Japanese display.

(2) After completion of setting, a guide message “Power

OFF and restart system.” is indicated. Power off the
instrument and then restart it. Its language will be
changed.

Power OFF and restart system.

2 - 32
 3.1

3. FIRST MEASUREMENT
3.1 What this Product can Do

Determination of Solution Concentration:

This product can measure the absorbance of a solution and
determine the concentration from the measured value.
Automatic continuous measurement → section 3.2.1.
Sample-by-sample measurement → section 3.3.1.

Measurement of Absorbance/Transmittance:
This product can measure the absorbance and transmittance of
a solution at a maximum of 6 wavelengths.
Automatic continuous measurement → section 3.2.2.
Sample-by-sample measurement → section 3.3.2.

Estimation of DNA Purity:

This product can measure the sample absorbance (at 230 nm,
260 nm, 280 nm and 320 nm) and calculate the absorbance ratio
(A260/A280 or A260/A230) for DNA purity check.
Automatic continuous measurement → section 3.2.3.
Sample-by-sample measurement → section 3.3.3.

Measurement of Spectrum:
This product can measure the absorption spectrum or
transmission spectrum of a sample.
Measurement after automatic baseline
Correction → section 3.2.4.
Sample-by-sample measurement → section 3.3.4.

Measurement of Change with Time:

This product can measure the absorbance or
transmittance of a sample at a specified wavelength. →section
3.3.5.

CAUTION
Fatigue due to Long-hour Operation
Operating the instrument while watching the display in the
same posture for long hours will cause fatigue of the eyes
and body. When working with the display for a long time,
take a break for 10 to 15 minutes every hour to relax your
eyes and body.

3-1
3.2 Automatic Continuous Measurement (6-cell auto mode)

3.2 Automatic Continuous Measurement (6-cell auto mode)

Up to 6 samples can be measured automatically (auto zero

sample is contained; spectrum measurement is excluded.).
Automatic absorbance zero correction or baseline correction is
possible by using the position of cell A.

3
4 2

5 1
A
1 2 3 4 5

Determination of Solution Concentration → section 3.2.1.

Generation of calibration curve and
determination of unknown sample concentration → section
3.2.1
Determination of unknown sample concentration
by entered calibration curve factor → section 3.2.1.
Determination of unknown sample concentration
from saved calibration curve → section 4.1.4.
Measurement of Absorbance/Transmittance →section 3.2.2.
Measurement of DNA Purity →section 3.2.3.
Measurement of Spectrum →section 3.2.4.

3-2
 3.2.1

3.2.1 Determining Concentration of Solution

This instrument is used for generating a calibration curve and

determining the concentration of an unknown sample, and for
entering a calibration curve factor and determining the
concentration of an unknown sample.

Guide: When auto start is specified, the system automatically

sets up the conditions and advances to the
measurement screen phase of sequence after the
power switch is turned on. For how to specify auto
start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3..)

2. Setting of Measuring Conditions

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 3-1) will appear. To set each condition for
quantitative calculation, press the <1> key
(Photometry) and then [ENTER] key. Or select
“Photometry” with the [▲] or [▼] key and press the
[ENTER] key.

Main Menu CELL A

^1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
^5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 3-1 Main Menu Screen

(2) The Photometry screen (Fig. 3-2) appears. To set the

measuring conditions, press the <1> key (Meas.
PARAM.) and then [ENTER] key. Or select “Meas.
PARAM.” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

3-3
3.2 Automatic Continuous Measurement (6-cell auto mode)

Photometry CELL A
1 Meas.PARAM Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _

Fig. 3-2 Photometry Screen

(3) The measuring condition (Meas. PARAM.) screen

(Fig. 3-3) appears. To select concentration
measurement (CONC) in Data Mode, press the <1>
key (CONC) and then [ENTER] key. After that, with
the [▲] or [▼] key, set the number of wavelengths (No.
of WL), wavelength (WL) and initial delay (Delay). For
details of each parameter, refer to Table 3-1 and
Explanation 3-1.

Photometry CELL A
1 Meas.PARAM. Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _
□1 CONC □
2 ABS □
3 %T □4 DNA

Fig. 3-3 Measuring Condition (Meas. PARAM.)

Screen

3-4
 3.2.1

Table 3-1 “Meas. PARAM.” Setting Parameters

Setting Item Description

Data Mode For measuring the concentration of a solution, select
[1] CONC.
[1] CONC
[2] ABS : Used for measuring the absorbance.
(For details, refer to section 3.2.2..)
[3] %T : Used for measuring the transmittance.
(For details, refer to section 3.2.2..)
[4] DNA : Used for estimating the purity of DNA.
(For details, refer to section 3.2.3..)
No. of WL Set the number of wavelengths to be used for
measurement. Normally specify “1”. When
subtracting background absorption, specify “2” or “3”.
(→ For details of setting, refer to Explanation 3-1.)
WL1 (nm) to Enter a wavelength to be used for measurement.
WL3 (nm) This parameter is settable in a range of 190.0 to
1100.0 nm in steps of 0.1 nm.
When “1” is set for No. of WL:
Enter a wavelength to be used for measurement for
WL1.
When “2” is set for No. of WL:
For WL1, enter a wavelength at which background
absorption is obtained. For WL2, enter a
wavelength at which absorption deriving from the
target substance for quantitative determination is
obtained. In so doing, it is required that WL1 > WL2.
When “3” is set for No. of WL:
For WL1 and WL3, enter a wavelength at which
background absorption is obtained. For WL3, enter
a wavelength at which absorption deriving from the
target substance for quantitative determination is
obtained. In so doing, it is required that WL1 >
WL2 > WL3.
Delay (s) After the [START] key is pressed, the system waits
for the time period set here and then starts
measurement. This parameter is settable in a range
of 0 to 9999 sec in steps of 1 sec.
Set this parameter when you want to start
measurement after a specified time period, including
when measuring a sample after its temperature
reaches the room temperature or to start
measurement after completion of reaction.
Enter “0” if the delay time need not be set.

3-5
3.2 Automatic Continuous Measurement (6-cell auto mode)

Explanation 3-1 Setting of Number of Wavelengths

Number of
Description
Wavelengths
1 Most generally used. Specify a wavelength for WL1. A calibration curve is
generated from obtained absorbance A and the concentration of an unknown
sample is determined.
A = A(1)

A(1)
Absorbance

WL1
Wavelength
Absorption spectrum of unknown sample

2 Effective when the background is flat. For WL2, specify a wavelength at which
absorption deriving from the target substance for quantitative determination is
obtained and for WL1, specify a wavelength at which background absorption is
obtained. Absorbance A is calculated by the equation given below replacing the
obtained absorbance values with A(2) and A(1). A calibration curve is generated
from the value thus calculated and the concentration of an unknown sample is
determined.
A = A(2)－A(1)

A(2)
Absorbance

A(1)
A(1)
WL2 WL1
Wavelength
Absorption spectrum of unknown sample

3 Effective When a sample is turbid or the background is not flat. Absorbance A is

calculated by the equation given below replacing the absorbance values at 3
wavelengths with A(1), A(2) and A(3). A calibration curve is generated from the
value thus calculated and the concentration of an unknown sample is determined.
(WL1 − WL2) × A(3) + (WL 2 − WL3) × A(1)
A = A(2) −
WL1 − WL3
(where, WL1＞WL2＞WL3)
A(2)

A(3)
Absorbance

A(1)

WL3 WL2 WL1

Wavelength
Absorption spectrum of unknown sample

3-6
 3.2.1

(4) After completion of setting, return to the Photometry

screen by pressing the [RETURN] key or [◄] key.

3. Setting of Sample Conditions

(1) To set the sample conditions, press the <2> key

(Sample) and then [ENTER] key. Or select “Sample”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The sample condition (Sample) screen (Fig. 3-4)

appears.

Photometry CELL A
1 Meas.PARAM. Sample Name
2 Sample Sample ID 1
3 Curve Statistics OFF
4 Curve Data
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Input character. ：_
Sample Name [Max: 8 characters]
ABC

Fig. 3-4 Sample Condition (Sample) Screen

(3) To enter/select the sample conditions, press the

[ENTER] key. Or select each item with the [▲] or [▼]
key and make your setting. For details of each
parameter, refer to Table 3-2.

(4) After setting the sample name (Sample Name), sample

number (Sample ID), statistical calculation (Statistics)
and number of calculations (No. of Calc), return to the
Photometry screen by pressing the [RETURN] key or
[◄] key.

3-7
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-2 “Sample” Setting Parameters

Setting Item Description

Sample Up to 8 characters can be entered for the sample
Name name. The sample name entered here is printed in
the sample name field on the report. For the
character input procedure, refer to section 2.4.3.
Sample ID Set the first number to be assigned for sample
numbering.
This parameter is settable in a range of 1 to 9999.
If “9999” is specified for Sample ID, the samples are
numbered 9999 → 1 → 2 ....
Statistics Specify whether to perform statistical calculation.
1) ON : Statistical calculation is carried out.
2) OFF : Statistical calculation is not carried out.
In statistical calculation, the mean value (MEAN),
standard deviation (SD) and relative standard
deviation (RSD) are calculated according to the
equations given below for the quantitative values of
samples. This calculation is carried out for every
number of calculations (N) set by the following
parameter.
[Mean value]
N

∑X i
MEAN = i =1

N
(N = Number of calculations)

[Standard deviation]
2
⎛ N 2⎞ ⎛ N ⎞
⎜∑ X i ⎟ − ⎜∑ Xi ⎟ N
SD = ⎝ i =1 ⎠ ⎝ i =1 ⎠
N −1
(N = Number of calculations)

[Relative standard deviation]

SD
RSD = × 100
MEAN
No. of Calc This parameter is indicated when ON is specified for
Statistics. Set the number of samples for statistical
calculation. The number is settable in a range of 2
to 100. If “3” is set for No. of Calc, statistical
calculation is carried out for every 3 samples.

3-8
 3.2.1

4. Setting of Calibration Curve Conditions

(1) To set the calibration curve conditions, press the <3>

key (Curve) and then [ENTER] key. Or select “Curve”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The calibration curve condition (Curve) screen (Fig. 3-

5) appears.

Photometry CELL A
1 Meas.PARAM. Curve Type 1st Order
2 Sample Thru Zero ON
3 Curve No.of STD 3
4 Curve Data Upper LMT． 9999
5 6 Cell Lower LMT. 0.000
6 System Unit Label mg/l
7 Print
8 Save Method
9 Meas.Screen
Select function. : _
□1 1st Order □
2 Factor

Fig. 3-5 Calibration Curve Condition (Curve)

Screen

(3) To enter/select the calibration curve conditions, press

the [ENTER] key. Or select each item by pressing the
[▲] or [▼] key and make your setting. For details of
each parameter, refer to Table 3-3.

(4) After setting the calibration curve type (Curve Type),

through-zero function (Through Zero), upper
concentration limit (Upper LMT.), lower concentration
limit (Lower LMT.) and concentration unit (Unit Label),
return to the Photometry screen by pressing the
[RETURN] key or [◄] key.

3-9
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-3 “Curve” Setting Parameters

Setting Item Description

Curve Type Select a method for calculating the concentration of
an unknown sample from the following two.
1) 1st Order : Standards (standard solutions) are
measured, regression analysis
(least-squares method) is made
based on the relationship between
the standard concentrations and
obtained absorbance to generate a
calibration curve. The concentration
of an unknown sample is determined
from the calibration curve thus
obtained.
[Calculation formula]
For the calculation formula for the calibration
curve factor, refer to Explanation 3-2.
2) Factor : The slope and intercept of a
calibration curve are entered and the
concentration of an unknown sample
is determined based on its factor.
This method is used for determining
the concentration according to the
literature or calibration curve data
measured with other instrument, or
for calculating the concentration by
using the molar absorption coefficient
or ratio absorbance to multiply the
absorbance by factor.
[Calculation formula]
• When ABS = f(CONC) is selected for Curve
(calibration curve formula) on System screen
Y = A1･X +A0
X = (Y - A0) / A1
• When CONC = f(ABS) is selected for Curve
(calibration curve formula) on System screen
X = A1･Y +A0
where, X : Quantitative value of unknown
sample
Y : Absorbance of unknown sample
(measurement result)
A0, A1 : Input values
Thru Zero Select generation of a calibration curve which is
forced through the origin (concentration 0,
absorbance 0) or generation of a calibration curve
which is not forced through the origin.
1) ON : A calibration curve which is forced through
the origin is generated.
2) OFF : A calibration curve which is not forced
through the origin is generated.

3 - 10
 3.2.1

(cont’d)
Setting Item Description
Thru Zero When ON is selected, regression analysis is made
taking the intercept of the calibration curve as zero.
When OFF is selected, regression analysis is made
with the intercept of the calibration curve.
For calculation of the regression formula, refer to
Explanation 3-2.
For details of usage, refer to Explanation 3-3 and
Explanation 3-4.
No. of STD Set the number of standards (standard solutions) to
be measured in a range of 1 to 20.
Note that “1” is settable only when ON is selected for
Thru Zero.
Upper LMT. If the result of quantitative determination is larger than
the set value, “HI” is printed at the side of the result.
This indication is used for readily checking if the
result is within the normal concentration range.
This parameter is settable in a range of 0 to 9999.
Lower LMT. If the result of quantitative determination is smaller
than the set value, “LO” is printed at the side of the
result. This indication is used for readily checking if
the result is within the normal concentration range.
This parameter is settable in a range of 0 to 9999.
Unit Label Up to 8 characters can be entered for the unit of
concentration. (mg/L, %, mol/L and ppm for
example)
For the character input procedure, refer to section
2.4.3.

3 - 11
3.2 Automatic Continuous Measurement (6-cell auto mode)

Explanation 3-2 How to Calculate Regression Formula

Calibration Curve
Formula
Thru Zero ON Thru Zero OFF
(system
condition)
Abs = f(Conc.) Regression formula: Y = A1・X Regression formula: Y = A1・X +A0

A1 =
∑X Y n n
A0 =
(∑ X )(∑ Y ) − (∑ X )(∑ X Y )
n
2
n n n n

∑X n(∑ X ) − (∑ X )
2
2 2
n
n n

n(∑ X nYn ) − (∑ X n )(∑ Yn )

A1 =
( )
n ∑ X n − (∑ X n )
2 2

Conc. = f(Abs) Regression formula: X = A1・Y Regression formula: X = A1・Y +A0

A1 =
∑Y X n n
A0 =
(∑ Y )(∑ X ) − (∑ Y )(∑ Y X )
n
2
n n n n

∑Y n(∑ Y ) − (∑ Y )
2
2 2
n
n n

n(∑ Yn X n ) − (∑ Yn )(∑ X n )
A1 =
( )
n ∑ Yn − (∑ Yn )
2 2

X: Standard concentration
Y: Obtained standard absorbance
n : Number of standards

5. Setting of Calibration Curve Data

(1) To set the calibration curve data, press the <4> key
(Curve Data) and then [ENTER] key. Or select
“Curve Data” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

[When Curve - Curve Type: 1st Order is selected]

(2) The calibration curve data (Curve Data) screen (Curve

Type: 1st Order) (Fig. 3-6) appears.

3 - 12
 3.2.1

Photometry CELL A
1 Meas.PARAM. STD CONC
2 Sample 1 0.000
3 Curve 2 10.00
4 Curve Data 3 20.00
5 System
6 Print
7 Save Method
8 6 Cell
9 Meas.Screen
Input value. ： ＿
STD1 [0.000 - 9999]

Fig. 3-6 Calibration Curve Data (Curve Data)

Screen (Curve Type: 1st Order)

(3) Enter the concentration of each standard (standard

solution). After that, press the [ENTER] key.
You can enter the concentration of each standard by
using the [▲] or [▼] key. Here, key in the
concentration values according to the number of
standards entered on the calibration curve condition
(Curve) screen. The concentration can be entered in
a range of 0.000 to 9999.

[When Curve - Curve Type: Factor is selected]

(2)’ The calibration curve data (Curve Data) screen (Curve

Type: Factor) (Fig. 3-7) appears.

Photometry CELL A
1 Meas.PARAM. A0 0.0000
2 Sample A1 1.0000
3 Curve
4 Curve Data
5 System
6 Print
7 Save Method
8 6 Cell
9 Meas.Screen
Input value. ： ＿
A0 [±99999 – 0.0000]

Fig. 3-7 Calibration Curve Data (Curve Data)

Screen (Curve Type: Factor)

(3)’ Enter calibration curve factors. After that, press the

[ENTER] key. You can enter each factor by using the
[▲] or [▼] key. The factor can be entered in 5
significant digits in a range of 0.0000 to ±99999.
For how to set the calibration curve factors, refer to
Table 3-4.

3 - 13
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-4 How to Set Calibration Curve Factors

[Normal Usage]
Calibration Curve Formula (system Calibration Curve Formula (system
condition): condition):
ABS = f(CONC) CONC = f(ABS)
Calculation Absorbance = A1 × Concentration + Concentration = A1 × Absorbance +
formula A0 A0
Concentration = (Absorbance - A0)/A1
Use of literature
value or Absorbance Concentration
calibration curve (Y axis) (Y axis)
data measured
with other Concentration (X axis) Absorbance (X axis)
instrument
Enter A1: Slope value and A0: Y- Enter A1: Slope value and A0: Y-
intercept value with the absorbance on intercept value with the concentration
the Y axis and the concentration on the on the Y axis and the absorbance on
X axis. the X axis.
Use of molar When the molar absorption coefficient When the molar absorption coefficient
absorption is ε (M-1cm-1), the optical path length of is ε (M-1cm-1), the optical path length of
coefficients cell is L (cm) and the concentration is cell is L (cm) and the concentration is
C (µM), Absorbance = εCL/1000. C (µM), Absorbance = εCL/1000.
For determining concentration C, enter For determining concentration C, enter
A1: εL/1000 and A0: 0. A1: 1000/εL and A0: 0.
Use of ratio When the molar absorption coefficient When the molar absorption coefficient
absorbance is E1%1cm, the optical path length of cell is E1%1cm, the optical path length of cell
is L (cm) and the concentration is C is L (cm) and the concentration is C
(mg/L), Absorbance = E1%1cmCL/10000. (mg/L), Absorbance = E1%1cmCL/10000.
For determining concentration C, enter For determining concentration C, enter
A1: E1%1cmL/10000 and A0: 0. A1: 10000/E1%1cmL and A0: 0.

(4) After setting each standard concentration or calibration

curve factors, return to the Photometry screen by
pressing the [RETURN] key or [◄] key.

6. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <5> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

3 - 14
 3.2.1

(2) The 6-cell condition (6 Cell) screen (Fig. 3-8) appears

(Curve Type: 1st Order).

Photometry CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample STD Autozero STD1
3 Curve SAMP.Autozero ON
4 Curve Data Autozero int. 5
5 6 Cell No.of Sample 1
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _
□1 Auto □
2 Manual

Fig. 3-8 6-cell Condition (6 Cell) Screen

(3) To set each item on the 6-cell condition (6 Cell) screen,

press the [ENTER] key. Or select each item with the
[▲] or [▼] key and make your setting according to the
guidance. For details of each parameter, refer to
Table 3-5.

(4) After selection/input of the 6-cell mode (6 Cell Mode),

calibration curve auto zero (STD Autozero), sample
auto zero (SAMP. Autozero), auto zero interval
(Autozero int.) and number of samples (No. of Sample),
return to the Photometry screen by pressing the
[RETURN] key or [◄] key.

Table 3-5 “6 Cell” Setting Parameters

Setting Item Description

6 Cell Mode Specify a 6-cell operation mode for measurement.
1) Auto
In this mode, a sample is set at each position of the
6-cell holder beforehand and the holder is
automatically turned to automatically perform from
auto zero to standard/sample measurement. It is
recommended to use this mode if you have 6 cells
and there are many samples to be measured.
2) Manual
In this mode, standard/sample measurement is
carried out by using one position (cell 1 to 5
selectable) of the 6-cell holder. To eliminate the
influence of instrument drift (baseline fluctuation)
during measurement, auto zero can easily be
executed by setting a solution for auto zero to cell A.
In normal usage, set a standard or sample solution to
cell 1. (For details of the manual mode, refer to
section 2.5.4..)

3 - 15
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
Setting Item Description
Curve Select a sample to be used for auto zero (operation
Autozero for adjusting the absorbance to zero) in standard
(indicated measurement between STD1 and Blank.
only when 1) STD1
1st Order is
selected for Selected when the auto zero sample in standard
Curve Type) measurement is a standard solution whose
concentration is 0 (STD1). In standard
measurement, auto zero is automatically performed
with STD1. With STD1 selected, the absorbance is
adjusted to 0 with a solution whose concentration is
0. It is therefore recommended to specify 1) ON for
Thru Zero in Table 3-3.
2) Blank
Selected when the auto zero sample in standard
measurement is other than a standard solution whose
concentration is 0 (blank). In standard
measurement, auto zero is automatically performed
with this blank solution. Normally, select 2) OFF set
for Thru Zero in Table 3-3.
* For details of setting, refer to Explanation 3-3 and
Explanation 3-4.
SAMP. You can select a sample for auto zero in sample
Autozero measurement or specify avoidance of automatic
execution of auto zero.
1) ON
Selected when the auto zero sample in sample
measurement is the same as the one in standard
measurement. Auto zero is automatically carried
out.
2) Sample Blank
Selected when the auto zero sample in sample
measurement is a sample blank. Auto zero is
automatically carried out with the sample blank.
Used for calculating the quantitative value by
subtracting the sample blank absorbance from the
sample absorbance.
3) OFF
Auto zero is not carried out automatically in sample
measurement. You can carry out auto zero
manually.
* For details of setting, refer to Explanation 3-3 and
Explanation 3-4.
Autozero int. 1) 5
Auto zero is automatically carried out for every 5
measurements.
2) 1
Auto zero is automatically carried out for each
sample.
No. of Set the number of samples to be measured.
Sample The number is settable in a range of 1 to 150.

3 - 16
 3.2.1

Explanation 3-3 How to Set 6-cell Measurement Parameters without

Coloring Reagent

Standard solution Sample solution

Absorbance

Analyte of interest Absorbance of analyte

Absorbance-zero line by
Solvent auto zero with <1>
Cell

<1> <2> <3> <4>

STD1 STD2 STD3 Sample

Elements causing absorbance without using coloring reagent (not containing interfering substance)

A measured absorbance value is given as a total of absorptions due to various elements (cell,
solvent, coloring reagent, analyte of interest, interfering substance). In other words, this value
does not simply stand for an absorption due to the analyte alone.
For generating a calibration curve according to concentration levels 0, 1 and 2, solutions of
concentration levels 0, 1 and 2 are prepared for STD1, STD2 and STD3, respectively (see <1> to
<3> in the above figure). These STD’s have absorptions due to cell, solvent and analyte.
However, the absorbance required for actual quantitation is the one deriving from the analyte.
Therefore, absorptions due to cell and solvent need to be subtracted (by auto zero operation).
In this case, auto zero is carried out with STD 1 and the absorbance values of STD1 to STD3 are
measured, thereby generating a calibration curve.
And, for sample quantitation, the concentration of the sample is determined using the absorbance
after exclusion of cell and solvent. When the absorption other than those due to cell and solvent
derives from the analyte alone, the determined concentration equals the concentration of the
analyte as illustrated at <4> in the above figure. For quantitation of this sample with 6 Cell Mode
set to Auto, the STD Autozero, SAMP. Autozero and Thru Zero are set to STD1, ON or OFF and
ON, respectively. (Refer to the measurement parameters in “A” of the following table.)

Standard solution Sample solution

Absorbance

Interfering Absorbance of analyte

Analyte of interest
substance
Absorbance-zero line by
auto zero with <5>
Absorbance-zero line by
Solvent auto zero with <1>
Cell

<1> <2> <3> <5> <4>

STD1 STD2 STD3 Blank test Sample
Solution (interfering substance contained)

Elements causing absorbance without using coloring reagent (containing interfering substance)

3 - 17
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
In the case where a sample contains an interfering substance such as turbid or disturbing
component as illustrated at <6> in the above figure, the absorbance due to the interfering substance
will be added to the quantitative value of the analyte.
In this case, it is necessary to carry out a blank test (the same preparatory operation as for samples
by use of pure water or the like) and subtract the measured absorbance of the blank test solution by
auto-zero operation at the time of sample measurement or subtract the quantitative value of the
blank test solution from that of the sample in order to subtract the absorbance of the interfering
substance from that of the sample. For quantitation of the sample with 6 Cell Mode set to Auto and
subtraction by auto zero, the STD Autozero, SAMP. Autozero and Thru Zero are set to STD1,
SAMP. BLK and ON, respectively. (Refer to the measurement parameters in “B” of the following
table.)

Setting of Each Measurement Parameter without Coloring Reagent

A: Usual Method (in case B: Correction with Blank Test

of sample <4>) Solution (in case of sample <6>)
Application
For Subtracting Absorbance of
Common Usage
Blank Test Solution
STD Autozero STD1 STD1
SAMP. Autozero ON OFF Sample Blank
Thru Zero ON ON

3 - 18
 3.2.1

Explanation 3-4 How to Set 6-cell Measurement Parameters with Coloring Reagent

Standard solution Sample solution

Absorbance

Analyte of interest Absorbance of analyte

Coloring reagent

Absorbance-zero line by
Solvent auto zero with <1>
Cell

<0> <1> <2> <3> <4>

Blank STD1 STD2 STD3 Sample
(solvent and
cell)

Elements causing absorbance with coloring reagent (not containing interfering substance)

When using a coloring reagent, the absorption of a sample solution includes that of this reagent
besides those of cell and solvent. For subtracting the absorbance due to the coloring reagent as
well, auto zero is carried out with the solution at <1> in the above figure.
For sample quantitation, the concentration of the sample is determined using the absorbance after
exclusion of cell, solvent and coloring reagent. When the absorption other than those due to cell,
solvent and coloring reagent derives from the analyte alone, the determined concentration equals
the concentration of the analyte as illustrated at <4> in the above figure. For quantitation of this
sample with 6 Cell Mode set to Auto, the STD Autozero, SAMP. Autozero and Thru Zero are set to
STD1, ON or OFF and ON, respectively. (Refer to the measurement parameters in “D” of the
following table.)
Also, when the absorbance of the solution at <1> in the above figure is unstable with time, the
solution is not suited for auto zero operation. Therefore, it is recommended to carry out auto zero
with the blank (sample). In this case, STD Autozero, SAMP. Autozero and Thru Zero are set to
Blank, ON or OFF and ON, respectively. (Refer to the measurement parameters in “E” of the
following table.)

Standard solution Sample solution

Absorbance of analyte
Absorbance

Analyte of Interfering
interest substance Absorbance-zero line by
auto zero with <5>

Coloring Absorbance-zero line by

reagent auto zero with <1>
Solvent
Cell

<0> <1> <2> <3> <5> <4>

Blank STD1 STD2 STD3 Blank test Sample
(solvent and Solution (interfering substance contained)
cell)

3 - 19
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
In the case where a sample contains an interfering substance such as turbid or disturbing
component as illustrated at <6> in the above figure, the absorbance due to the interfering substance
will be added to the quantitative value of the analyte. In this case, it is necessary to carry out a
blank test (the same preparatory operation as for samples by use of pure water or the like) and
subtract the measured absorbance of the blank test solution by auto-zero operation at the time of
sample measurement or subtract the quantitative value of the blank test solution from that of the
sample in order to subtract the absorbance of the interfering substance from that of the sample.
For quantitation of the sample with 6 Cell Mode set to Auto and subtraction by auto zero, the STD
Autozero, SAMP. Autozero and Thru Zero are set to STD1, Sample Blank and ON, respectively.
(Refer to the measurement parameters in “F” of the following table.)

Setting of Each Measurement Parameter with Coloring Reagent

D: Usual Method E: Auto Zero with F: Correction with Blank

(in case of Blank (in case of Test Solution (in case of
sample <4>) sample <4>) sample <6>)
Application When STD1
Common Usage Absorbance is For Subtracting Absorbance
(auto zero with Unstable with Time of Blank Test Solution
STD1) (auto zero with
blank)
STD Autozero STD1 Blank STD1
SAMP. Autozero ON OFF ON OFF Sample Blank
Thru Zero ON OFF ON

3 - 20
 3.2.1

7. Setting of System Conditions

(1) To set the system conditions, press the <6> key

(System) and then [ENTER] key. Or select “System”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The system condition (System) screen (Fig. 3-9)

appears. Set the condition for the calibration curve
regression formula according to the guidance.
For detail, refer to Table 3-6.

Photometry CELL A
1 Meas.PARAM. Curve ABS=f(CONC)
2 Sample
3 Curve
4 Curve Data
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _
□1 ABS=f(CONC) □
2 CONC=f(ABS)

Fig. 3-9 System Condition (System) Screen

Table 3-6 “System” Setting Parameter

Setting Item Description

Curve Select a format for the calibration curve formula
between the following two.
1) ABS = f(CONC)
The calibration curve formula is expressed in the
format of (Absorbance = A1 × Concentration + A0).
This format is normally used.
2) CONC = f(ABS)
The calibration curve formula is expressed in the
format of (Concentration = A1 × Absorbance + A0).
This format is used only when the reference
calibration curve is expressed in the format of CONC
= f(ABS), or when Factor is selected for Curve Type
and the value obtained by adding a numerical value
to the absorbance multiplied by factor is taken as a
concentration.

(3) After setting the calibration curve factor (Factor), return to

the Photometry screen by pressing the [RETURN] key or
[◄] key.

3 - 21
3.2 Automatic Continuous Measurement (6-cell auto mode)

8. Setting of Printing Conditions

GUIDE: Specify print items if determined beforehand.

You can also specify print items after completion
of measurement. If you do not want to set the
printing conditions here, proceed to step 9.

(1) To set the printing conditions, press the <7> key (Print)
and then [ENTER] key. Or select “Print” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The printing condition (Print) screen (Fig. 3-10)

appears.

Photometry CELL A
1 Meas.PARAM. Print Date ON
2 Sample Run Date ON
3 Curve Method ON
4 Curve Data Curve ON
5 6 Cell STD ON
6 System Sample ON
7 Print
8 Save Method
9 Meas.Screen
Select function. : _
□1 ON □
2 OFF

Fig. 3-10 Printing Condition (Print) Screen

(3) To select the printing conditions, press the [ENTER]

key. Or select each item with the [▲] or [▼] key and
make your setting according to the guidance.
For details of each parameter, refer to Table 3-7.

(4) After specifying whether to print out the printing date

(Print Date), analysis date (Run Date), conditions
(Method), calibration curve (Curve), standard data
(STD) and sample data (Sample), return to the
Photometry screen by pressing the [RETURN] key or
[◄] key.

3 - 22
 3.2.1

Table 3-7 “Print” Setting Parameters

Location in
Setting Item Description
Fig. 3-11
Print Date [1] ON : Printing date is printed. <1>
[2] OFF : Printing date is not printed.
Run Date [1] ON : Analysis date is printed. <2>
[2] OFF : Analysis date is not printed.
Method [1] ON : Measuring conditions are <3>
printed.
[2] OFF : Measuring conditions are
not printed.
Curve [1] ON : Calibration curve is printed. <4>
[2] OFF : Calibration curve is not
printed.
STD [1] ON : Results of standard <5>
measurement are printed.
[2] OFF : Results of standard
measurement are not
printed.
Sample [1] ON : Results of sample <6>
measurement are printed.
[2] OFF : Results of sample
measurement are not
printed.

3 - 23
3.2 Automatic Continuous Measurement (6-cell auto mode)

Report：2010/04/10 15:12 ……….. <1>

0.800

<4>

0.000
CONC(mg/l)
0.000 1.000

Sample Name ：
File Name ：
Run Date ： 2010/04/10 13:34 ……….. <2>
Operator ：

Spectrophotometer
Model： U-5100 Spectrophotometer
Serial No.： 1234-567
ROM VER.： 3J25300
Option：

Instrument
Meas. Mode： Photometry Bandpass(nm)： 5.0
Data Mode： CONC 6 Cell Mode： Auto
No.of WL ： 1 STD Autozero： STD1
WL1(nm)： 540.0 SAMP.Autozero： ON <3>
Delay(s)： 0 Autozero int.： 5
No.of Sample： 3

STD
ID ABS CONC(mg/l) DIFF RD T
STD1 0.001 0.000 0.000 -0.024 -0.026
STD2 0.008 0.010 0.000 -0.060 -0.065
STD3 0.033 0.040 0.000 -0.053 -0.058
STD4 0.086 0.100 -0.004 -1.538 -1.663 <5>
STD5 0.169 0.200 -0.004 -1.498 -1.621
STD6 0.413 0.500 0.000 -0.189 -0.205
STD7 0.659 0.800 0.002 0.689 0.745

Curve Type： 1st Order

Curve： ABS = f(CONC)
Thru Zero： ON
Lower LMT.： 0.000
Upper LMT．： 0.800 <3>
Curve Factor： A0= 0.0000 A1= 0.8261
Factor： Correlation Coef.:R= 1.0000 Determination Coef.:R2= 0.9999

Sample
ID ABS CONC(mg/l)
1 0.001 0.000
2 0.008 0.010 <6>
3 0.033 0.040

Fig. 3-11 Example of Printout in Photometry Mode

Setting of measuring conditions has now been completed.

For saving the set conditions, proceed to step 9. If they need
not be saved, then proceed to step 10.

3 - 24
 3.2.1

9. Saving of Measuring Conditions

GUIDE: When the set measuring conditions need not be

saved, proceed to step 10.

(1) To save the set measuring conditions, press the <8>

key (Save Method) and then [ENTER] key. Or select
“Save Method” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(2) The measuring condition saving (Save Method) screen

(Fig. 3-12) appears.

Save Method CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

File name Measurement Saving

mode date

File Name：_
File Name [Max: 20 characters]
ABC W X ( 1/ 1)

Fig. 3-12 Measuring Condition Saving (Save

Method) Screen (with saved data)

(3) When other measuring conditions are saved in

advance, a list of saved measuring conditions appears
on the main menu. At this time, the file name,
measurement mode (PHT: Photometry, WLS:
Wavelength Scan, TMS: Time Scan) and saving date
are indicated.

(4) For the measuring conditions to be saved, enter a file

name within 20 characters and press the [ENTER] key.

3 - 25
3.2 Automatic Continuous Measurement (6-cell auto mode)

NOTE:
• If a file of the same name as entered already exists in
the same measurement mode, the following guidance
appears.
Already exists. Overwrite it? ：_＿
1 Yes
□ 2 No
□

For overwriting, press the <1> key (Yes). To avoid

overwriting, press the <2> key (No) and rename the file
to be saved.
• A total of 50 method files can be saved for all
measurement modes.
• If 50 files are already registered, the following message
appears.
No.of files is full.

In this case, delete files referring to section 4.2.2 and

retry saving.
• The method for which auto start (for details, refer to
section 4.5) is set is preceded by “*”.

(5) Return to the Photometry screen by pressing the

[RETURN] key.

10. Measurement of Standard Solutions

GUIDE: When 2) Factor is selected for Curve Type in

step 4, standard solution measurement is not
carried out. So proceed to step 11.

(1) To determine the measuring conditions set in the

preceding steps, press the <9> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key.

(2) The standard setting screen (Fig. 3-13) appears.

3 - 26
 3.2.1

Photometry CELL A
[Info]
Set STD to Cell No..
Cell A STD1 (0.000)
Cell number Cell 1 -
Cell 2 STD2 (10.00) Standard
Cell 3 STD3 (20.00) concentration

Standard name Example of display

START:Meas.: _ with “3” set for
number of
standards

Fig. 3-13 Standard Setting Screen

On the screen, displayed from the left are the cell

number, standard name and standard concentration.
Set the corresponding standard to each cell. If “-”
(hyphen) is indicated for the standard name, avoid
setting a standard in the relevant cell. When STD1 is
set for “STD Autozero”, set the cells as shown in Table
3-8. The instrument measures standards in the
sequence shown in Table 3-9. When Blank is set for
“STD Autozero”, set the cells as shown in Table 3-10.
The instrument measures standards in the sequence
shown in Table 3-11. Measurement is carried out for
every 5 standards until the number set in “No. of STD”
is reached.

Table 3-8 How to Set Cells with STD Selected for

STD Autozero

Cell A Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round STD1 Not set STD2 STD3 STD4 STD5
2nd round STD1 STD6 STD7 STD8 STD9 STD10
3rd round STD1 STD11 STD12 STD13 STD14 STD15
4th round STD1 STD16 STD17 STD18 STD19 STD20

3 - 27
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-9 Auto Zero Interval and Operation of Instrument (1)

Sequence of Operation
Cell Position Operation
Auto Zero Interval: 5 Auto Zero Interval: 1
Setting of standards (1st round)
1 1 Cell A Auto zero
2 2 Cell A STD1 measurement
⎯ 3 Cell A Auto zero
3 4 Cell 2 STD2 measurement
⎯ 5 Cell A Auto zero
4 6 Cell 3 STD3 measurement
⎯ 7 Cell A Auto zero
5 8 Cell 4 STD4 measurement
⎯ 9 Cell A Auto zero
6 10 Cell 5 STD5 measurement
Setting of standards (2nd round)
7 11 Cell A Auto zero
8 12 Cell 1 STD6 measurement
⎯ 13 Cell A Auto zero
9 14 Cell 2 STD7 measurement
Repeated until the specified number of standards are all measured.

Table 3-10 How to Set Cells with Blank Selected for

STD Autozero

Cell A Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Blank STD1 STD2 STD3 STD4 STD5
2nd round Blank STD6 STD7 STD8 STD9 STD10
3rd round Blank STD11 STD12 STD13 STD14 STD15
4th round Blank STD16 STD17 STD18 STD19 STD20

3 - 28
 3.2.1

Table 3-11 Auto Zero Interval and Operation of Instrument (2)

Sequence of Operation
Cell Position Operation
Auto Zero Interval: 5 Auto Zero Interval: 1
Setting of standards (1st round)
1 1 Cell A Auto zero
2 2 Cell 1 STD1 measurement
⎯ 3 Cell A Auto zero
3 4 Cell 2 STD2 measurement
⎯ 5 Cell A Auto zero
4 6 Cell 3 STD3 measurement
⎯ 7 Cell A Auto zero
5 8 Cell 4 STD4 measurement
⎯ 9 Cell A Auto zero
6 10 Cell 5 STD5 measurement
Setting of standards (2nd round)
7 11 Cell A Auto zero
8 12 Cell 1 STD6 measurement
⎯ 13 Cell A Auto zero
9 14 Cell 2 STD7 measurement
Repeated until the specified number of standards are all measured.

(3) Upon completion of setting, press the [START] key.

Measurement will then start.

(4) During measurement, the screen shown in Fig. 3-14 is

displayed.

Photometry 600.0nm 0.099ABS CELL A

ID ABS CONC
STD1 0.001 0.000
STD2 10.00
STD3 20.00
STD4 30.00
STD5 40.00
Example of display
Meas.STD2...
with “3” set for
number of
standards

Fig. 3-14 Screen Displayed during STD2 Measurement

GUIDE: To stop measurement, press the [STOP] key.

To restart measurement, press the [START] key.

3 - 29
3.2 Automatic Continuous Measurement (6-cell auto mode)

(5) When “6” or more is specified for “No. of STD”, the

results obtained from measurement of STD1 to STD5
are indicated as shown in Fig. 3-15. This screen also
appears after completion of 2nd/3rd-round
measurement. Pressing the [START] key displays the
next standard setting screen. Pressing the <1> key
(Curve Confirmation) or <2> key (Remeasure) allows
the processing or remeasurement shown in Table 3-12.

Photometry CELL A
ID ABS CONC
STD1 0.001 0.000
STD2 0.099 10.00
STD3 0.203 20.00
STD4 0.300 30.00
STD5 0.405 40.00

START:Continue; Select function.:_＿

□1 Curve Confirmation □2 Remeasure

Fig. 3-15 Example of Display after 1st-round

Standard Measurement

Table 3-12 Guidance during Standard Measurement

Setting Item Description

[1] Curve You can generate a calibration curve with the
Confirmation standards measured before now and confirm it.

Photometry/Curve CELL A
R=0.9999 R2=0.9999
0.405
+
+
ABS
+
+
0.001+
0.000 CONC 40.00
Select function. : _
□1 Scale □2 Factor □
3 STD

[1] Scale : Allows you to change the scale of

calibration curve.
[2] Factor : Indicates the factor, correlation
coefficient and determination
coefficient of calibration curve.
[3] STD : Indicates the result of standard
solution measurement.
[2] Remeasure You can remeasure the last-measured standard.

3 - 30
 3.2.1

(6) When the specified number of standards have all been

measured, the screen shown in Fig. 3-16 appears.
When you select the <1> key (Sample Meas.), the
system terminates standard measurement and
proceeds to sample measurement. Selecting the <2>
key (Curve), <3> key (Save) and <4> key (Print) allows
the processings shown in Table 3-13.

Photometry CELL A

ID ABS CONC
STD6 0.498 50.00
STD7 0.602 60.00
STD8 0.707 70.00
STD9 0.800 80.00
STD10 0.893 90.00

Select function. : _
□1 Sample Meas. □
2 Curve □
3 Save □
4 Print

Fig. 3-16 Guidance after Standard Measurement

(No. of STD: 10)

Table 3-13 Guidance after Standard Measurement

Setting Item Description

[1] Sample The system proceeds to sample measurement.
Meas.
[2] Curve You can generate a calibration curve with the
measured standards and confirm it. When this
function is selected, the following screen appears.

Photometry/Curve CELL A
R=0.9999 R2=0.9999
0.893 +
+
+
+
ABS +
+
+
0.001+ +
0.000 CONC 90.00
Select function. : _
□1 Scale □
2 Factor □
3 STD

3 - 31
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
Setting Item Description
[2] Curve [1] Scale : Allows you to change the scale of
calibration curve.
[2] Factor : Indicates the factor, correlation
coefficient and determination coefficient
of calibration curve.
[3] STD : Indicates the result of standard solution
measurement. Also allows
remeasurement of a standard solution,
deletion of standard data unnecessary
for generation of a calibration curve or
retrieval of deleted data.
[3] Save You can save the measured standard data.
When this function is selected, the following screen
appears.

Save Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
□1 Save □
2 Sort
W X ( 1/ 1)

[1] Save : Allows you to save the result data under

a new file name.
[2] Sort : Allows you to rearrange the presently
saved data in the order of file names or
dates.
[4] Print You can select items to be printed. When this
function is selected, the screen given below appears.
For each item, specify whether to print out (ON or
OFF).

Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Curve ON
STD ON
Sample ON

Select function. : _
□1 ON □
2 OFF
W X ( 1/ 1)

3 - 32
 3.2.1

11. Measurement of Sample Solutions

(1) The sample setting screen (Fig. 3-17) appears.

Photometry CELL A
[Info]
Set samples to Cell No..
Cell A STD1 (0.000)
Cell 1 Sample1
Cell 2 Sample2
Cell 3 Sample3
Cell 4 Sample4
Cell 5 Sample5

START:Meas.:

Fig. 3-17 Sample Setting Screen

On the screen, displayed from the left are the cell

number and sample name. Set the corresponding
sample to each cell. If “-” (hyphen) is indicated for the
sample name, avoid setting a sample in the relevant
cell. When ON is selected for “SAMP. Autozero”, set
the samples as shown in Table 3-14, as shown in
Table 3-15 when Sample Blank is selected and as
shown in Table 3-16 when OFF is selected.
The instrument measures samples in the sequence
shown in Table 3-17.
When OFF is set for “SAMP. Autozero”, auto zero can
be executed by pressing the [AUTOZERO] key when
the guidance on sample setting is indicated.
For details, refer to Fig. 3-18.

3 - 33
3.2 Automatic Continuous Measurement (6-cell auto mode)

<1> Press the [AUTOZERO] key when the sample setting screen is
displayed.

Photometry CELL A
[Info]
Set samples to Cell No..
Cell A -
Cell 1 Sample1
Cell 2 Sample2
Cell 3 Sample3
Cell 4 Sample4
Cell 5 Sample5

START:Meas.:＿

Sample setting screen

<2> The auto zero execution screen appears. Set an auto zero
sample to cell A. The lamp is lit for 30 seconds and the current
measured value is indicated so that you can check if execution
of auto zero is necessary.

Photometry 600.0nm 0.001ABS CELL A

[Info]

Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Auto zero execution screen

<3> Press the [AUTOZERO] key. Auto zero is executed and the
sample setting screen reappears.

Fig. 3-18 Manual Auto Zero with OFF Set for

SAMP. Autozero

Upon completion of sample setting according to the

guidance, press the [START] key. Measurement will
then start. Measurement is carried out for every 5
samples. Guidance is given until the specified
number of samples (No. of Sample on 6 Cell screen)
are all measured.

GUIDE: For how to set a sample to the cell holder, refer

to section 2.4.4..

3 - 34
 3.2.1

Table 3-14 How to Set Cells with ON Selected for SAMP. Autozero

Cell A* Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round STD1 or blank Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
2nd round STD1 or blank Sample 6 Sample 7 Sample 8 Sample 9 Sample 10
3rd round STD1 or blank Sample 11 Sample 12 Sample 13 Sample 14 Sample 15
4th round and Repeated until the specified number of samples are all measured.
subsequent

* Set the solution selected in “Curve Autozero”. When Factor is selected for Curve Type, set STD1
or blank.

Table 3-15 How to Set Cells with Sample Blank Selected for SAMP. Autozero

Cell A Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Sample blank Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
2nd round Sample blank Sample 6 Sample 7 Sample 8 Sample 9 Sample 10
3rd round Sample blank Sample 11 Sample 12 Sample 13 Sample 14 Sample 15
4th round and Repeated until the specified number of samples are all measured.
subsequent

Table 3-16 How to Set Cells with OFF Selected for SAMP. Autozero

Cell A* Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Not set STD1 STD2 STD3 STD4 STD5
2nd round Not set STD6 STD7 STD8 STD9 STD10
3rd round Not set STD11 STD12 STD13 STD14 STD15
4th round and Repeated until the specified number of samples are all measured.
subsequent

* Upon completion of each round, you can execute auto zero with cell A if needed. For details,
refer to Fig. 3-18.

3 - 35
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-17 Auto Zero Interval and Operation of Instrument

SAMP. Autozero: ON
SAMP. Autozero: OFF Auto Zero Auto Zero Cell Position Operation
Interval: 5 Interval: 1
Setting of samples (1st round)
⎯ 1 1 Cell A Auto zero
1 2 2 Cell 1 Sample 1 measurement
⎯ ⎯ 3 Cell A Auto zero
2 3 4 Cell 2 Sample 2 measurement
⎯ ⎯ 5 Cell A Auto zero
3 4 6 Cell 3 Sample 3 measurement
⎯ ⎯ 7 Cell A Auto zero
4 5 8 Cell 4 Sample 4 measurement
⎯ ⎯ 9 Cell A Auto zero
5 6 10 Cell 5 Sample 5 measurement
Setting of samples (2nd round)
⎯ 7 11 Cell A Auto zero
6 8 12 Cell 1 Sample 6 measurement
⎯ ⎯ 13 Cell A Auto zero
7 9 14 Cell 2 Sample 7 measurement
Repeated until the specified number of samples are all measured.

(2) During measurement, the screen shown in Fig. 3-19 is

displayed.

Photometry 600.0nm 0.101ABS CELL A

ID ABS CONC
1

Meas.Sample2...

Fig. 3-19 Screen Displayed during Sample

Measurement

3 - 36
 3.2.1

(3) When “6” or more is specified for “No. of Sample”, the

results obtained from measurement of samples 1 to 5
are indicated as shown in Fig. 3-16.
This measurement result screen also appears after
completion of 2nd/3rd-round measurement.
Pressing the [START] key displays the next sample
setting screen. Set the next samples according to the
guidance. (For operation when the <1> key or <2>
key is pressed, refer to Table 3-12.)

Photometry CELL A
ID ABS CONC
1 0.001 0.000
2 0.101 10.01
3 0.410 40.90
4 0.300 30.00
5 0.205 20.50

START:Continue; Select function.: ＿

□1 Curve Confirmation □2 Remeasure

Fig. 3-20 Example of Display after 1st-round

Sample Measurement

(4) When the specified number of samples have all been

measured, the screen shown in Fig. 3-21 appears.
For the processing function assigned to each key, refer
to Table 3-18.

Photometry CELL A
ID ABS CONC
6 0.301 30.00
7 0.251 25.11
8 0.419 40.90
9 0.110 11.00
10 0.234 23.50

Select function. : _
□1 Meas. □
2 Curve □
3 Save □
4 Print

W X( 2/ 2)

Fig. 3-21 Example of Display after Sample

Measurement
(number of samples: 10)

3 - 37
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-18 Guidance after Sample Measurement

Setting Item Description

[1] Meas. You can make setting related to measurement.
New Meas., Continue and Remeasure are selectable.
[1] New Meas. : Selected when newly measuring
samples with the same calibration
curve.
[2] Continue : Allows you to start measurement
from the number following the
previously measured sample.
When this function is selected, the
guidance given below appears.
Enter the number of samples to be
measured and press the [ENTER]
key.

Input value. ： ＿
No.of Sample [1-140]

Display with 10 samples measured

[3] Remeasure : Selected when remeasuring a

measured sample. When this
function is selected, the guidance
given below appears. Select a
sample to be remeasured with the
[▲] or [▼] key, set the sample to
cell 1 and press the [START] key to
carry out remeasurement.
To execute auto zero, press the
[AUTOZERO] key.

Photometry CELL A
ID ABS CONC
6 0.301 30.00
7 0.251 25.11
8 0.419 40.90
9 0.110 11.00
10 0.234 23.50

Set sample to Cell1(Sample10).: _

START:Meas.; AUTOZERO:Autozero
W X( 2/ 2)

3 - 38
 3.2.1

(cont’d)
Setting Item Description
[2] Curve You can generate a calibration curve with the
measured standards and confirm it. When this
function is selected, the following screen appears.

Photometry/Curve CELL A
R=0.9999 R2=0.9999
0.893 +
+
+
+
ABS +
+
+
0.001+ +
0.000 CONC 90.00
Select function. : _
□1 Scale □
2 Factor □
3 STD

For details of this function, refer to section 4.3.

[1] Scale : Allows you to change the scale of
calibration curve.
[2] Factor : Indicates the factor, correlation
coefficient and determination coefficient
of calibration curve.
[3] STD : Indicate the result of standard solution
measurement. Also allows
remeasurement of a standard solution,
deletion of a standard unnecessary for
generating a calibration curve and
retrieval of deleted data.
[3] Save You can save the measured data. When this
function is selected, the following screen appears.

Save Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
□1 Save □
2 Sort
W X ( 1/ 1)

[1] Save : Allows you to save data under a new file

name.
[2] Sort : Allows you to rearrange the currently
saved data in the order of file names or
dates.

3 - 39
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
Setting Item Description
[4] Print You can select items to be printed. When this
function is selected, the screen given below appears.
For each item, select whether to print out (ON or
OFF).

Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Curve ON
STD ON
Sample ON

Select function. : _
□1 ON □
2 OFF

3 - 40
 3.2.2

3.2.2 Measuring Absorbance/Transmittance

This instrument is capable of measuring the absorbance and

transmittance of a solution at a maximum of 6 wavelengths.

GUIDE: When auto start is specified, the system automatically

sets up the conditions and advances to the
measurement screen phase of sequence after the
power switch is turned on. For how to specify auto
start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3..)

2. Setting of Measuring Conditions

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 3-22) will appear.

(2) To set each condition for quantitative calculation, press

the <1> key (Photometry) and then [ENTER] key.
Or select “Photometry” with the [▲] or [▼] key and
press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 3-22 Main Menu Screen

3 - 41
3.2 Automatic Continuous Measurement (6-cell auto mode)

(3) The Photometry screen (Fig. 3-23) appears.

Photometry CELL A
1 Meas.PARAM Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _

Fig. 3-23 Photometry Screen

(4) To set the measuring conditions, press the <1> key

(Meas. PARAM.) and then [ENTER] key. Or select
“Meas. PARAM.” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(5) The measuring condition (Meas. PARAM.) screen (Fig.

3-24) appears.

Photometry CELL A
^1 Meas.PARAM. Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 6 Cell
6 System
^7 Print
8 Save Method
9 Meas.Screen
Select function. : _
□1 CONC □2 ABS □
3 %T □4 DNA

Fig. 3-24 Measuring Condition (Meas. PARAM.)

Screen

(6) For Data Mode, press the <2> key (ABS) for
absorbance measurement or <3> key (%T) for
transmittance measurement and then press the
[ENTER] key.

(7) With the [▲] or [▼] key, set the number of

wavelengths (No. of WL) and wavelength (WL).
For details of each parameter, refer to Table 3-19.

3 - 42
 3.2.2

(8) After completion of setting, return to the Photometry

screen by pressing the [RETURN] key or [◄] key.

Table 3-19 “Meas. PARAM.” Setting Parameters

Setting Item Description

Data Mode For measuring the absorbance, select 2) ABS.
For measuring the transmittance, select 3) %T.
1) CONC : Used for generating a calibration curve
and determining the concentration.
(For details, refer to section 3.2.1..)
2) ABS : Used for measuring the absorbance.
3) %T : Used for measuring the transmittance.
4) DNA : Used for estimating the purity of DNA.
(For details, refer to section 3.2.3..)
No. of WL Set the number of wavelengths to be used for
measurement.
This parameter is selectable in a range of 1 to 6.
WL1 (nm) to Enter a wavelength to be used for measurement.
WL6 (nm) This parameter is settable in a range of 190.0 to
1100.0 nm in steps of 0.1 nm.
Delay (s) After the [START] key is pressed, the system waits
for the time period set here and then starts
measurement. This parameter is settable in a range
of 0 to 9999 sec in steps of 1 sec.
Set the parameter when you want to start
measurement after a specified time period, including
when measuring a sample after its temperature
reaches the room temperature and when starting
measurement after completion of reaction. Enter “0”
if the delay time need not be set.

3. Setting of Sample Conditions

(1) To set the sample conditions, press the <2> key

(Sample) and then [ENTER] key. Or select “Sample”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The sample condition (Sample) screen (Fig. 3-25)

appears.

3 - 43
3.2 Automatic Continuous Measurement (6-cell auto mode)

Photometry CELL A
1 Meas.PARAM. Sample Name
2 Sample Sample ID 1
3 6 Cell
4 Print
5 Save Method
6 Meas.Screen

Input character. ： ＿
Sample Name [Max: 8 characters]

Fig. 3-25 Sample Condition (Sample) Screen

(3) To enter/select the sample conditions, press the

[ENTER] key. Or select each item with the [▲] or [▼]
key and make your setting. For details of each
parameter, refer to Table 3-20.

(4) After setting the sample name (Sample Name) and

sample number (Sample ID), return to the Photometry
screen by pressing the [RETURN] key or [◄] key.

Table 3-20 “Sample” Setting Parameters

Setting Item Description

Sample Up to 8 half-size alphanumeric characters can be
Name entered for the sample name. The sample name
entered here is printed in the sample name field on
the report. For the character input procedure, refer
to section 2.4.3.
Sample ID Set the first number to be assigned for sample
numbering.
This parameter is settable in a range of 1 to 9999.
If “9999” is specified for Sample ID, the samples are
numbered 9999 → 1 → 2 ....

4. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

3 - 44
 3.2.2

(1) To set the 6-cell conditions, press the <3> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The 6-cell condition (6 Cell) screen (Fig. 3-26) appears.

Photometry CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample SAMP.Autozero ON
3 6 Cell Autozero int. 5
4 Print No.of Sample 1
5 Save Method
6 Meas.Screen

Select function. : _
□1 Auto □
2 Manual

Fig. 3-26 6-cell Condition (6 Cell) Screen

(3) To set each item on the 6-cell condition (6 Cell) screen,

press the [ENTER] key. Or select each item with the
[▲] or [▼] key and make your setting according to the
guidance. For details of each parameter, refer to
Table 3-21.

(4) After selecting/entering the 6-cell mode (6 Cell Mode),

sample auto zero (SAMP. Autozero), auto zero interval
(Autozero int.) and number of samples (No. of Sample),
return to the Photometry screen by pressing the
[RETURN] key or [◄] key. Setting of measuring
conditions has now been completed. For saving the
set conditions, proceed to step 6. If they need not be
saved, then proceed to step 7.

3 - 45
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-21 “6 Cell” Setting Parameters

Setting Item Description

6 Cell Mode Specify a 6-cell operation mode for measurement.
1) Auto
In this mode, a sample is set at each position of the
6-cell holder beforehand and the holder is
automatically turned to automatically perform from
auto zero to standard/sample measurement. It is
recommended to use this mode if you have 6 cells
and there are many samples to be measured.
2) Manual
In this mode, standard/sample measurement is
carried out by using one position (cell 1 to 5
selectable) of the 6-cell holder. To eliminate the
influence of instrument drift (baseline fluctuation)
during measurement, auto zero can easily be
executed by setting a solution for auto zero to cell A.
In normal usage, set a standard or sample solution to
cell 1. (For details of the Manual mode, refer to
section 3.3.2..)
SAMP. You can select an auto zero sample in sample
Autozero measurement or specify avoidance of automatic
execution of auto zero.
1) ON
Auto zero is automatically carried out in sample
measurement. The auto zero interval is set in
Autozero int.
2) OFF
Auto zero is not carried out automatically in sample
measurement. You can carry out auto zero
manually.
Autozero int. 1) 5
Auto zero is automatically carried out for every 5
measurements.
2) 1
Auto zero is automatically carried out for each
sample.
No. of Set the number of samples to be measured.
Sample This parameter is settable in a range of 1 to 150.

3 - 46
 3.2.2

5. Setting of Printing Conditions

GUIDE: Specify print items if determined beforehand.

You can also specify print items after completion
of measurement. If you do not want to set the
printing conditions here, proceed to step 6.

(1) To set the printing conditions, press the <4> key (Print)
and then [ENTER] key. Or select “Print” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The printing condition (Print) screen (Fig. 3-27)

appears.

Photometry CELL A
1 Meas.PARAM. Print Date ON
2 Sample Run Date ON
3 6 Cell Method ON
4 Print Sample ON
5 Save Method
6 Meas.Screen

Select function. : _
□1 ON □
2 OFF

Fig. 3-27 Printing Condition (Print) Screen

(3) To select the printing conditions, press the [ENTER]

key. Or select each item with the [▲] or [▼] key and
make your setting according to the guidance. For
details of each parameter, refer to Table 3-22.

(4) After specifying whether to print out the printing date

(Print Date), analysis date (Run Date), conditions
(Method) and sample data (Sample), return to the
Photometry screen by pressing the [RETURN] key or
[◄] key.

3 - 47
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-22 “Print” Setting Parameters

Location in
Setting Item Description
Fig. 3-28
Print Date [1] ON : Printing date is printed. <1>
[2] OFF : Printing date is not printed.
Run Date [1] ON : Analysis date is printed. <2>
[2] OFF : Analysis date is not printed.
Method [1] ON : Measuring conditions are <3>
printed.
[2] OFF : Measuring conditions are
not printed.
Sample [1] ON : Results of sample <4>
measurement are printed.
[2] OFF : Results of sample
measurement are not
printed.

Report：2010/04/10 15:12 ……….. <1>

Sample Name：
File Name：
Run Date： 2010/04/10 13:34 ……….. <2>
Operator：

Spectrophotometer
Model： U-5100 Spectrophotometer
Serial No.： 1234-567
ROM VER.： 3J25300
Option：

Instrument
Meas.Mode： Photometry Bandpass(nm)： 5.0
Data Mode： ABS 6 Cell Mode： Auto
No.of WL： 1 SAMP.Autozero： ON <3>
WL1(nm)： 540.0 Autozero int.： 5
Delay(s)： 0 No.of Sample： 3

Sample
ID ABS
1 0.001 <4>
2 0.008
3 0.033

Fig. 3-28 Example of Printout in Absorbance/Transmittance

Measurement

3 - 48
 3.2.2

6. Saving of Measuring Conditions

GUIDE: When the set measuring conditions need not be

saved, proceed to step 7.

(1) To save the set measuring conditions, press the <5>

key (Save Method) and then [ENTER] key. Or select
“Save Method” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(2) The measuring condition saving (Save Method) screen

(Fig. 3-29) appears.

Meas.PARAM. CELL A
1 CheckB (at 500nm) PHT 2010/10/10
2 Quality Check PHT 2010/03/30
3 ABS meas.(at 200nm) PHT 2010/04/29

File name Measurement Saving

mode date

File Name：_
File Name [Max: 20 characters]
W X ( 1/ 1)

Fig. 3-29 Measuring Condition Saving (Save

Method) Screen

(3) When other measuring conditions are saved in

advance, a list of saved measuring conditions appears
on the main menu. At this time, the file name,
measurement mode (PHT: Photometry, WLS:
Wavelength Scan, TMS: Time Scan) and saving date
are indicated.

(4) For the measuring conditions to be saved, enter a file

name within 20 characters and press the [ENTER] key.

3 - 49
3.2 Automatic Continuous Measurement (6-cell auto mode)

NOTE:
• If a file of the same name as entered already exists in
the same measurement mode, the following guidance
appears.
Already exists. Overwrite it? :_
1 Yes □
□ 2 No

For overwriting, press the <1> key (Yes). To avoid

overwriting, press the <2> key (No) and rename the file
to be saved.
• A total of 50 method files can be saved for all
measurement modes.
• If 50 files are already registered, the following message
appears.
No.of files is full.

In this case, delete files referring to section 4.2.2 and

retry saving.
• The method for which auto start (for details, refer to
section 4.5.) is set is preceded by “*”.

(5) Return to the Photometry screen by pressing the

[RETURN] key.

3 - 50
 3.2.2

7. Measurement of Samples

(1) To determine the measuring conditions set in the

preceding steps, press the <6> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key. The sample setting screen (Fig. 3-30)
appears.

Photometry CELL A
[Info]
Set samples to Cell No..
Cell A Blank
Cell 1 Sample1
Cell 2 Sample2
Cell 3 Sample3
Cell 4 Sample4
Cell 5 Sample5

START:Meas.:

Fig. 3-30 Sample Setting Screen

On the screen, displayed from the left are the cell

number and sample name. Set the corresponding
sample to each cell. If you do not know what sample
should be set to cell A (blank), refer to Explanation 3-5
in absorbance measurement and Explanation 3-6 in
transmittance measurement.

3 - 51
3.2 Automatic Continuous Measurement (6-cell auto mode)

Explanation 3-5 Method of Auto Zero in Absorbance Measurement

Cell A
Absorbance

Substance B
Absorbance of analyte
Substance A

Absorbance-zero line in
auto zero with cell and solvent
Solvent
Cell
<1> <2> <3>
Cell and Sample 1 Sample 2
solvent

Auto Zero for Measuring Absorbance Except for Cell and Solvent

A measured absorbance is given as a total absorbance of various elements (such as cell, solvent,
substances contained in sample). Therefore, the analyst must extract necessary absorbance from
the total absorbance.
Assume the following have been prepared; a solvent contained in cell <1> and 2 sample solutions;
sample solution 1 containing substance A in cell <2> and sample solution 2 containing substances
A and B in cell <3>. Reference should be made to the above figure ([Auto Zero for Measuring
Absorbance except for Cell and Solvent]). We see that the absorbance of the solution in cell <2>
derives from the cell, solvent and substance A, and that of the solution in cell <3> derives from the
cell, solvent and substances A and B. For targeting the absorbance except for cell and solvent,
auto zero is carried out with the cell containing a solvent alone. As a result, the absorbance of only
the substance A is measurable with <2> and a sum of the absorbance values due to substances A
and B is measurable with <3>.

Cell A
Absorbance

Substance B Absorbance of analyte

Substance A
Absorbance-zero line in
auto zero with sample 1

Solvent
Cell
<1> <2> <3>
Cell and Sample 1 Sample 2
solvent

Auto Zero for Measuring Absorbance of Substance B

For measuring the absorbance of only the substance B, auto zero operation is carried out with
sample 1 in cell <2>. By this operation, the absorbance of only the substance B in cell <3> can be
measured. [Refer to “Auto Zero for Measuring Absorbance of Substance B” in the above figure.]
For extracting the absorbance of an analyte of interest, the absorbance due to elements other than
the analyte can be subtracted by performing auto zero operation with a solution having a
composition which does not contain the analyte. The solution subjected to the auto zero operation
is called a “blank” in absorbance/transmittance measurement. You should select an optimum
blank in consideration of its role explained here.

3 - 52
 3.2.2

Explanation 3-6 Method of Auto Zero in Transmittance Measurement

Cell A

Transmittance 100% Line in

Auto Zero with Air
Transmittance

Transmit-
tance
100%
Transmittance of
analyte
<1> <2> <3>
Air Sample 1 Sample 2

Auto Zero with Transmittance of Air Set at 100%

In transmittance measurement, it is important what condition is to be defined as transmittance

100%. For measuring the transmittance of a sample with the transmittance in air assumed to be
100%, auto zero operation is carried out with nothing contained in cell A except for air. Refer to
the above figure (“Auto Zero with Transmittance of Air Set at 100%”). Then, by measuring
samples 1 and 2, their transmittance values with respect to that of air will be obtainable.

Cell A

Transmittance 100% Line in

Auto Zero with Sample 1
Transmittance

Transmittance
100%

Transmittance of
analyte

<1> <2> <3>

Air Sample 1 Sample 2

Auto Zero with Transmittance of Cell and Solvent Set at 100%

For transmittance measurement on assumption that the transmittance of sample 1 is 100%, auto
zero operation is carried out with this sample contained in cell A. By this method, the
transmittance of sample 2 will be obtainable with respect to that of sample 1. The sample
subjected to the auto zero operation is called a “blank” in absorbance/transmittance measurement.
You should select an optimum blank in consideration of its role explained here.

3 - 53
3.2 Automatic Continuous Measurement (6-cell auto mode)

If “-” (hyphen) is indicated for the sample name, avoid

setting a sample in the relevant cell. When ON is
selected for “SAMP. Autozero”, set samples as shown
in Table 3-23, and as shown in Table 3-24 when OFF
is selected. The instrument measures samples in the
sequence shown in Table 3-25.

Table 3-23 How to Set Cells with ON Selected for SAMP. Autozero

Cell A* Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Blank Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
2nd round Blank Sample 6 Sample 7 Sample 8 Sample 9 Sample 10
3rd round Blank Sample 11 Sample 12 Sample 13 Sample 14 Sample 15
4th round and Repeated until the specified number of samples are all measured.
subsequent

Table 3-24 How to Set Cells with OFF Selected for SAMP. Autozero

Cell A* Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Not set STD1 STD2 STD3 STD4 STD5
2nd round Not set STD6 STD7 STD8 STD9 STD10
3rd round Not set STD11 STD12 STD13 STD14 STD15
4th round and Repeated until the specified number of samples are all measured.
subsequent

* Upon completion of each round, you can execute auto zero with cell A if needed. For details,
refer to Fig. 3-31.

3 - 54
 3.2.2

Table 3-25 Auto Zero Interval and Operation of Instrument

SAMP. Autozero: ON
SAMP. Autozero: OFF Auto Zero Auto Zero Cell Position Operation
Interval: 5 Interval: 1
Setting of samples (1st round)
⎯ 1 1 Cell A Auto zero
1 2 2 Cell 1 Sample 1 measurement
⎯ ⎯ 3 Cell A Auto zero
2 3 4 Cell 2 Sample 2 measurement
⎯ ⎯ 5 Cell A Auto zero
3 4 6 Cell 3 Sample 3 measurement
⎯ ⎯ 7 Cell A Auto zero
4 5 8 Cell 4 Sample 4 measurement
⎯ ⎯ 9 Cell A Auto zero
5 6 10 Cell 5 Sample 5 measurement
Setting of samples (2nd round)
⎯ 7 11 Cell A Auto zero
6 8 12 Cell 1 Sample 6 measurement
⎯ ⎯ 13 Cell A Auto zero
7 9 14 Cell 2 Sample 7 measurement
Repeated until the specified number of samples are all measured.

When OFF is set for “SAMP. Autozero”, auto zero can

be executed by pressing the [AUTOZERO] key when
the guidance on sample setting is indicated.
For details, refer to Fig. 3-31.

3 - 55
3.2 Automatic Continuous Measurement (6-cell auto mode)

(1) Press the AUTOZERO key when the sample setting screen is
displayed.

Photometry CELL A
[Info]
Set samples to Cell No..
Cell A -
Cell 1 Sample1
Cell 2 Sample2
Cell 3 Sample3
Cell 4 Sample4
Cell 5 Sample5

START:Meas.:

Sample setting screen

(2) The auto zero execution screen appears. Set an auto zero
sample to cell A.

Photometry CELL A

[Info]

Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Auto zero execution screen

(3) Auto zero is executed and the sample setting screen reappears.

Fig. 3-31 Manual Auto Zero with OFF set for

SAMP. Autozero

Upon completion of sample setting according to the

guidance, press the [START] key. Measurement will
then start. Measurement is carried out for every 5
samples. Guidance is given until the specified
number of samples are all measured.

GUIDE: For how to set a sample to the cell holder, refer

to section 2.4.4.

3 - 56
 3.2.2

(2) During measurement, the screen shown in Fig. 3-32 is

displayed.

Photometry 600.0nm 0.101ABS CELL A

ID ABS
1 0.001
2 0.101
3

Meas.Sample3...

Fig. 3-32 Screen Displayed during Sample

Measurement

(3) When “6” or more is specified for “No. of Sample”, the

results obtained from measurement of samples 1 to 5
are indicated as shown in Fig. 3-33.
This measurement result screen also appears after
completion of 2nd/3rd-round measurement.
Pressing the [START] key displays the next sample
setting screen. Set the next samples according to the
guidance. (Pressing the <1> key (Remeasure) allows
remeasurement of the previous sample.)

Photometry CELL A
ID ABS
1 0.001
2 0.101
3 0.410
4 0.300
5 0.205

START:Continue; Select function.: ＿

1 Remeasure
□

Fig. 3-33 Example of Display after 1st-round

Sample Measurement

(4) When the specified number of samples have all been

measured, the screen shown in Fig. 3-34 appears.
For the processing function assigned to each key, refer
to Table 3-26.

3 - 57
3.2 Automatic Continuous Measurement (6-cell auto mode)

Photometry CELL A
ID ABS
6 0.301
7 0.251
8 0.419
9 0.110
10 0.234

Select function. : _
1)Meas. 2)Save 3)Print

Fig. 3-34 Example of Display after Sample

Measurement
(number of samples: 10)

Table 3-26 Guidance after Sample Measurement

Setting Item Description

Meas. You can make setting related to measurement.
“New Meas.”, “Continue” and “Remeasure” are
selectable.
[1] New Meas. : Selected when newly measuring
samples with the same calibration
curve.
[2] Continue : Allows you to start measurement
from the number following the
previously measured sample.
When this function is selected, the
guidance given below appears.
Enter the number of samples to be
measured and press the [ENTER]
key.

Input value. ： ＿
No.of Sample [1-140]

Display with 10 samples measured

[3] Remeasure : Selected when remeasuring a

measured sample. When this
function is selected, the guidance
given below appears. Select a
sample to be remeasured with the
[▲] or [▼] key, set the sample to
cell 1 and press the [START] key to
carry out remeasurement.
To execute auto zero, press the
[AUTOZERO] key.

3 - 58
 3.2.2

(cont’d)
Setting Item Description
Meas. Photometry CELL A
ID ABS CONC
6 0.301 30.00
7 0.251 25.11
8 0.419 40.90
9 0.110 11.00
10 0.234 23.50

Set sample to Cell1(Sample10).: _＿

START:Meas.; AUTOZERO:Autozero
W X( 2/ 2)

Save You can save the measured data. When this

function is selected, the following screen appears.

Save Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
□1 Save □
2 Sort
W X ( 1/ 1)

[1] Save : Allows you to save data under a new file

name.
[2] Sort : Allows you to rearrange the currently
saved data in the order of file names or
dates.
Print You can select items to be printed. When this
function is selected, the screen given below appears.
For each item, select whether to print out (ON or
OFF).

Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Sample ON

Select function. : _
□1 ON □
2 OFF

3 - 59
3.2 Automatic Continuous Measurement (6-cell auto mode)

3.2.3 Measuring DNA Sample (measurement through ratio calculation)

This instrument is capable of measuring the absorbance (at 230 nm, 260 nm, 280 nm and
320 nm) of a DNA sample and calculating the absorbance ratio (A260/A280 and A260/A230)
for check of the DNA purity. The instrument is also used for measuring the absorbance at 2
wavelengths and calculating the absorbance ratio or absorbance difference between the
values thus obtained.

Guide: When auto start is specified, the system automatically

sets up the conditions and advances to the
measurement screen phase of sequence after the
power switch is turned on. For how to specify auto
start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3..)

2. Setting of Measuring Conditions

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 3-35) will appear.

(2) To set each measuring condition, press the <1> key

(Photometry) and then [ENTER] key. Or select
“Photometry” with the [▲] or [▼] key and press the
[ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 3-35 Main Menu Screen

3 - 60
 3.2.3

(3) The Photometry screen (Fig. 3-36) appears.

Photometry CELL A
1 Meas.PARAM. Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 6 Cell
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _

Fig. 3-36 Photometry Screen

(4) To set the measuring conditions, press the <1> key

(Meas. PARAM.) and then [ENTER] key. Or select
“Meas. PARAM.” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(5) The measuring condition (Meas. PARAM.) screen (Fig.

3-37) appears.

Photometry CELL A
1 Meas.PARAM. Data Mode CONC
2 Sample No.of WL 1
3 Curve WL1(nm) 600.0
4 Curve Data Delay(s) 0
5 System
6 Print
7 Save Method
8 6 Cell
9 Meas.Screen
Select function. : _
□1 CONC □2 ABS □3 %T □
4 DNA

Fig. 3-37 Measuring Condition (Meas. PARAM.)

Screen

(6) For Data Mode, press the <4> key (DNA) and then
[ENTER] key to select DNA measurement.

(7) With the [▲] or [▼] key, set the number of pairs of
wavelengths (WL pair No.), background correction
(Bkgd. CORR.), wavelength (WL), correction
wavelength (CORR. WL) and initial delay time (Delay).
For details of each parameter, refer to Table 3-27.

(8) After completion of setting, return to the Photometry

screen by pressing the [RETURN] key or [◄] key.

3 - 61
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-27 “Meas. PARAM.” Setting Parameters

Setting Item Description

Data Mode For estimating the purity of DNA, select 4) DNA.
[1] CONC : Used for generating a calibration curve
and determining the concentration.
(For details, refer to section 3.2.1.)
[2] ABS : Used for measuring the absorbance.
(For details, refer to section 3.2.2.)
[3] %T : Used for measuring the transmittance.
(For details, refer to section 3.2.2.)
[4] DNA : Used for estimating the purity of DNA.
WL pair No. Set up the number of pairs of wavelengths to be used
for measurement.
Select whether to perform ratio calculation for one
pair of wavelengths or two pairs of wavelengths.
[1] WL pair No. 1 : Selected for ratio calculation for
one pair (either A260/A280
(absorbance ratio between 260
nm and 280 nm) or A260/A230).
[2] WL pair No. 2 : Selected for ratio calculation for
two pairs (both A260/A280 and
A260/A230).
Bkgd. Specify whether to perform background correction.
CORR. [1] ON : Selected when subtracting background
absorption.
[2] OFF: Selected when not subtracting
background absorption.
WL1 (nm) to Enter a wavelength to be used for measurement.
WL4 (nm) This parameter is settable in a range of 190.0 to
1100.0 nm in steps of 0.1 nm. The following
calculations are performed using the absorbance
values at the set wavelengths.
[WL pair No.: 1, Bkgd. CORR.: OFF]
Absorbance ratio = ABS (WL1)/ABS (WL2)
Absorbance difference = ABS (WL1) - ABS (WL2)
[WL pair No.: 1, Bkgd. CORR.: ON]
Absorbance ratio = (ABS (WL1) -
ABS (CORR. WL))/(ABS (WL2) - Data (CORR. WL))
Absorbance difference = ABS (WL1) - ABS (WL2)
[WL pair No.: 2, Bkgd. CORR.: OFF]
Absorbance ratio (WL: 1, 2) = ABS (WL1)/ABS (WL2)
Absorbance ratio (WL: 3, 4) = ABS (WL3) - ABS
(WL4)
[WL pair No.: 2, Bkgd. CORR.: ON]
Absorbance ratio (WL: 1, 2) = (ABS (WL1) -
ABS (CORR. WL))/(ABS (WL2) - ABS (CORR. WL))
Absorption ratio (WL: 3, 4) = (ABS (WL3) -
ABS (CORR. WL))/(ABS (WL4) - ABS (CORR. WL))
CORR. WL Set a wavelength to be used for background
correction. This parameter is indicated only when
ON is selected for Bkgd. CORR.
The parameter is settable in a range of 190.0 nm to
1100.0 in steps of 0.1 nm.

3 - 62
 3.2.3

(cont’d)
Setting Item Description
Delay (s) After the [START] key is pressed, the system waits
for the time period set here and then starts
measurement. This parameter is settable in a range
of 0 to 9999 sec in steps of 1 sec.
Set the parameter when you want to start
measurement after a specified time period, including
when measuring a sample after its temperature
reaches the room temperature. Enter “0” if the delay
time need not be set.

Explanation 3-7 Set Wavelengths in DNA Measurement

On the absorption spectrum of an aqueous nucleic acid solution, absorption has the minimum
point near 230 nm and the maximum point near 260 nm. The wavelength of maximum absorption
varies with the base content and sequence in the nucleic acid. It is known that the ratio of
A260/A230 is approximately 1.8 with DNA and 2.0 with RNA.
Also, because the coexisting protein have a maximum absorption point at 280 nm, sample purity
can be estimated by taking the ratio of A260/A280. When the ratio is smaller than 1.8 in the case
of DNA, protein may be coexistent. Conversely, it is known that the ratio becomes larger under the
coexistence of RNA.

Abs

1.5

1.0

0.5

0.0 nm
200 300 400
Absorption spectrum of salmon testis DNA

Excerpt from BUNSEKI KAGAKU BINRAN, 5th revised edition by The Japan Society of Analytical
Chemistry (page 400)

3 - 63
3.2 Automatic Continuous Measurement (6-cell auto mode)

Explanation 3-8 Background Correction

Bkgd.CORR Setting method

OFF Background correction is avoided. Absorbance ratio will be calculated using the
obtained absorbance values.
Absorbance ratio = A(1)/A(2)

Absorbance

A(1)
A(2)

WL2 WL1
Wavelength
Absorption spectrum

ON The background will be corrected. This is effective when the background level is
even along the wavelength axis. The absorbance at the set correction wavelength
will be subtracted from those at the measuring wavelengths. Calculation is carried
out according to the following equation.
Absorbance ratio = (A(1) - A(correction))(A(2) - A(correction))
Absorbance

A(1)
A(2)

A (correction)
WL2 WL1 Correction
wavelength
Wavelength
Absorption spectrum

3 - 64
 3.2.3

3. Setting of Sample Conditions

(1) To set the sample conditions, press the <2> key

(Sample) and then [ENTER] key. Or select “Sample”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The sample condition (Sample) screen (Fig. 3-38)

appears.

Photometry CELL A
1 Meas.PARAM. Sample Name 1
2 Sample Sample ID 1
3 6 Cell Statistics OFF
4 Print
5 Save Method
6 Meas.Screen

Input character. ： ＿
Sample Name [Max: 8 characters]
ABC

Fig. 3-38 Sample Condition (Sample) Screen

(3) To enter/select the sample conditions, press the

[ENTER] key. Or select each item with the [▲] or [▼]
key and make your setting. For details of each
parameter, refer to Table 3-28.

(4) After setting the sample name (Sample Name), sample

number (Sample ID) and statistical calculation
(Statistics), return to the Photometry screen by
pressing the [RETURN] key or [◄] key.

3 - 65
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-28 “Sample” Setting Parameters

Setting Item Description

Sample Up to 8 half-size alphanumeric characters can be
Name entered for the sample name. The sample name
entered here is printed in the sample name field on
the report. For the character input procedure, refer
to section 2.4.3.
Sample ID Set the first number to be assigned for sample
numbering.
This parameter is settable in a range of 1 to 9999.
If “9999” is specified for Sample ID, the samples are
numbered 9999 → 1 → 2 ....
Statistics For the sample absorbance ratio and absorbance
difference, the mean value (MEAN), standard
deviation (SD) and relative standard deviation (RSD)
are calculated according to the equations given
below. This calculation is carried out for every
number of calculations (N). The number of
calculations is set in the following setting item.
[Mean value]
N

∑X i
MEAN = i =1

N
(N = Number of calculations)

[Standard deviation]

2
⎛ N 2⎞ ⎛ N ⎞
⎜∑ Xi ⎟ − ⎜∑ Xi ⎟ N
SD = ⎝ i =1 ⎠ ⎝ i =1 ⎠
N −1

(N = Number of calculations)

[Relative standard deviation]

SD
RSD = × 100
MEAN

No. of Calc Set the number of samples for statistical calculation.

This parameter is indicated when ON is set for
Statistics. It is settable in a range of 2 to 100.
When “3” is set, statistical calculation is carried out
for every 3 samples.

3 - 66
 3.2.3

4. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <3> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The 6-cell condition (6 Cell) screen (Fig. 3-39) appears.

Photometry CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample SAMP.Autozero ON
3 6 Cell Autozero int. 5
4 Print No.of Sample 1
5 Save Method
6 Meas.Screen

Select function. : _
□1 Auto □2 Manual

Fig. 3-39 6-cell Condition (6 Cell) Screen

(3) To set each item on the 6-cell condition (6 Cell) screen,

press the [ENTER] key. Or select each item with the
[▲] or [▼] key and make your setting according to the
guidance. For details of each parameter, refer to
Table 3-29.

(4) After selecting/entering the 6-cell mode (6 Cell Mode),

sample auto zero (SAMP. Autozero), auto zero interval
(Autozero int.) and number of samples (No. of Sample),
return to the Photometry screen by pressing the
[RETURN] key or [◄] key.

3 - 67
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-29 “6 Cell” Setting Parameters

Setting Item Description

6 Cell Mode Specify a 6-cell operation mode for measurement.
[1] Auto
In this mode, a sample is set at each position of the
6-cell holder beforehand and the holder is
automatically turned to automatically perform from
auto zero to standard/sample measurement. It is
recommended to use this mode if you have 6 cells
and there are many samples to be measured.
[2] Manual
In this mode, standard/sample measurement is
carried out by using one position (cell 1 to 5
selectable) of the 6-cell holder. To eliminate the
influence of instrument drift (baseline fluctuation)
during measurement, auto zero can easily be
executed by setting a solution for auto zero to cell A.
In normal usage, set a standard or sample solution to
cell 1.
(For details of the Manual mode, refer to section
3.3.3..)
SAMP. You can select an auto zero sample in sample
Autozero measurement or specify avoidance of automatic
execution of auto zero.
[1] ON
Auto zero is automatically carried out in sample
measurement. The auto zero interval is set in
Autozero int.
[2] OFF
Auto zero is not carried out automatically in sample
measurement. You can carry out auto zero
manually.
Autozero int. [1] 5
Auto zero is automatically carried out for every 5
measurements.
[2] 1
Auto zero is automatically carried out for each
sample.
No. of Set the number of samples to be measured.
Sample This parameter is settable in a range of 1 to 150.

3 - 68
 3.2.3

5. Setting of Printing Conditions

GUIDE: Specify print items if determined beforehand.

You can also specify print items after completion
of measurement. If you do not want to set the
printing conditions here, proceed to step 6.

(1) To set the printing conditions, press the <4> key (Print)
and then [ENTER] key. Or select “Print” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The printing condition (Print) screen (Fig. 3-40)

appears.

Photometry CELL A
1 Meas.PARAM. Print Date ON
2 Sample Run Date ON
3 6 Cell Method ON
4 Print Sample ON
5 Save Method
6 Meas.Screen

Select function. : _
□1 ON □2 OFF

Fig. 3-40 Printing Condition (Print) Screen

(3) To select the printing conditions, press the [ENTER]

key. Or select each item with the [▲] or [▼] key and
make your setting according to the guidance.
For details of each parameter, refer to Table 3-30.

(4) After specifying whether to print out the printing date

(Print Date), analysis date (Run Date), conditions
(Method) and sample data (Sample), return to the
Photometry screen by pressing the [RETURN] key or
[◄] key.

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3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-30 “Print” Setting Parameters

Location in
Setting Item Description
Fig. 3-41
Print Date [1] ON : Printing date is printed. <1>
[2] OFF : Printing date is not printed.
Run Date [1] ON : Analysis date is printed. <2>
[2] OFF : Analysis date is not printed.
Method [1] ON : Measuring conditions are <3>
printed.
[2] OFF : Measuring conditions are
not printed.
Sample [1] ON : Results of sample <4>
measurement are printed.
[2] OFF : Results of sample
measurement are not
printed.

Report：2010/04/10 15:12 ……….. <1>

Sample Name：
File Name：
Run Date： 2010/04/10 13:34 ……….. <2>
Operator：

Spectrophotometer
Model： U-5100 Spectrophotometer
Serial No.： 1234-567
ROM VER.： 3J25300
Option：

Instrument
Meas.Mode： Photometry Bandpass(nm)： 5.0
Data Mode： DNA 6 Cell Mode： Auto
WL pair No.：
Bkgd.CORR.：
1
OFF
SAMP.Autozero：
Autozero int.：
ON
5
<3>
WL1(nm)： 260.0 No.of Sample： 3
WL2(nm)： 280.0
Delay(s) 0

Sample
ID WL1(nm) WL2(nm) ABS DIFF ABS Ratio
(260.0) (280.0) <4>
1 0.423 0.268 0.155 1.578
2 0.423 0.269 0.154 1.572

Fig. 3-41 Example of Printout in DNA Measurement

(5) Setting of measuring conditions has now been

completed. For saving the set conditions, proceed to
step 6. If they need not be saved, then proceed to
step 7.

3 - 70
 3.2.3

6. Saving of measuring conditions

GUIDE: When the set measuring conditions need not be

saved, proceed to step 7.

(1) To save the set measuring conditions, press the <5>

key (Save Method) and then [ENTER] key. Or select
“Save Method” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(2) The measuring condition saving (Save Method) screen

(Fig. 3-42) appears.

Meas.PARAM. CELL A
1 CheckB (at 500nm) PHT 2010/10/10
2 Quality Check PHT 2010/03/30
3 DNA meas. PHT 2010/04/29

File name Measurement Saving

mode date

File Name：_
File Name [Max: 20 characters]
W X ( 1/ 1)

Fig. 3-42 Measuring Condition Saving

(Save Method) Screen

(3) When other measuring conditions are saved in

advance, a list of saved measuring conditions appears
on the main menu. At this time, the file name,
measurement mode (PHT: Photometry, WLS:
Wavelength Scan, TMS: Time Scan) and saving date
are indicated.

(4) For the measuring conditions to be saved, enter a file

name within 20 characters and press the [ENTER] key.

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3.2 Automatic Continuous Measurement (6-cell auto mode)

NOTE:
• If a file of the same name as entered already exists in
the same measurement mode, the following guidance
appears.
Already exists. Overwrite it? :_
1) Yes 2) No

For overwriting, press the <1> key (Yes). To avoid

overwriting, press the <2> key (No) and rename the file
to be saved.
• A total of 50 method files can be saved for all
measurement modes.
• If 50 files are already registered, the following message
appears.
No.of files is full.

In this case, delete files referring to section 4.2.2 and

retry saving.
• The method for which auto start (for details, refer to
section 4.5.) is set is preceded by “*”.

(5) Return to the Photometry screen by pressing the

[RETURN] key.

7. Measurement of Samples

(1) To determine the measuring conditions set in the

preceding steps, press the <6> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key. The sample setting screen (Fig. 3-43)
appears.

Photometry CELL A

Set samples to Cell No..

Cell A Blank
Cell 1 Sample1
Cell number Cell 2 Sample2 Sample name
Cell 3 Sample3
Cell 4 Sample4
Cell 5 Sample5

START:Meas.:＿＿

Fig. 3-43 Sample Setting Screen

3 - 72
 3.2.3

On the screen, displayed from the left are the cell

number and sample name. Set the corresponding
sample to each cell. If “-” (hyphen) is indicated for the
sample name, avoid setting a sample in the relevant
cell. When ON is selected for “SAMP. Autozero”, set
samples as shown in Table 3-31, and as shown in
Table 3-32 when OFF is selected. The instrument
measures samples in the sequence shown in Table 3-
33.

Table 3-31 How to Set Cells with ON Selected for SAMP. Autozero

Cell A* Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Blank Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
2nd round Blank Sample 6 Sample 7 Sample 8 Sample 9 Sample 10
3rd round Blank Sample 11 Sample 12 Sample 13 Sample 14 Sample 15
4th round and Repeated until the specified number of samples are all measured.
subsequent

Table 3-32 How to Set Cells with OFF Selected for SAMP. Autozero

Cell A* Cell 1 Cell 2 Cell 3 Cell 4 Cell 5

1st round Not set STD1 STD2 STD3 STD4 STD5
2nd round Not set STD6 STD7 STD8 STD9 STD10
3rd round Not set STD11 STD12 STD13 STD14 STD15
4th round and Repeated until the specified number of samples are all measured.
subsequent

* Upon completion of each round, you can execute auto zero with cell A if needed. For details,
refer to Fig. 3-44.

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3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-33 Auto Zero Interval and Operation of Instrument

SAMP. Autozero: ON
SAMP. Autozero: OFF Auto Zero Auto Zero Cell Position Operation
Interval: 5 Interval: 1
Setting of samples (1st round)
⎯ 1 1 Cell A Auto zero
1 2 2 Cell 1 Sample 1 measurement
⎯ ⎯ 3 Cell A Auto zero
2 3 4 Cell 2 Sample 2 measurement
⎯ ⎯ 5 Cell A Auto zero
3 4 6 Cell 3 Sample 3 measurement
⎯ ⎯ 7 Cell A Auto zero
4 5 8 Cell 4 Sample 4 measurement
⎯ ⎯ 9 Cell A Auto zero
5 6 10 Cell 5 Sample 5 measurement
Setting of samples (2nd round)
⎯ 7 11 Cell A Auto zero
6 8 12 Cell 1 Sample 6 measurement
⎯ ⎯ 13 Cell A Auto zero
7 9 14 Cell 2 Sample 7 measurement
Repeated until the specified number of samples are all measured.

When OFF is set for “SAMP. Autozero”, auto zero can

be executed by pressing the [AUTOZERO] key when
the guidance on sample setting is indicated.
For details, refer to Fig. 3-44.

3 - 74
 3.2.3

<1> Press the [AUTOZERO] key when the sample setting screen is
displayed.

Photometry CELL A
[Info]
Set samples to Cell No..
Cell A -
Cell 1 Sample1
Cell 2 Sample2
Cell 3 Sample3
Cell 4 Sample4
Cell 5 Sample5

START:Meas.:＿

Sample setting screen

<2> The auto zero execution screen appears. Set an auto zero
sample to cell A. The lamp is lit for 30 seconds and the current
measured value is indicated so that you can check if execution
of auto zero is necessary.

Photometry 600.0nm 0.001ABS CELL A

[Info]

Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Auto zero execution screen

Fig. 3-44 Manual Auto Zero with OFF set for

SAMP. Autozero

Upon completion of sample setting according to the

guidance, press the [START] key. Measurement will
then start. Measurement is carried out for every 5
samples. Guidance is given until the specified
number of samples are all measured.

GUIDE: For how to set a sample to the cell holder, refer

to section 2.4.4.

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3.2 Automatic Continuous Measurement (6-cell auto mode)

(2) During measurement, the screen shown in Fig. 3-45 is

displayed.

Photometry 280.0nm 0.268ABS CELL A

ID ABS
3 WL1 （ 260.0nm）
WL2 （ 280.0nm）

ABS DIFF=
ABS Ratio=

Meas.Sample3...

Fig. 3-45 Screen Displayed during Sample

Measurement

(3) When “6” or more is specified for “No. of Sample”, the

results obtained from measurement of samples 1 to 5
are indicated as shown in Fig. 3-46. This
measurement result screen also appears after
completion of 2nd/3rd-round measurement.
Pressing the [START] key displays the next sample
setting screen. Set the next samples according to the
guidance. (Pressing the <1> key (Remeasure) allows
remeasurement of the previous sample.)

Photometry CELL A
ID ABS
3 WL1 （ 260.0nm） 0.423
WL2 （ 280.0nm） 0.268

ABS DIFF= 0.155

ABS Ratio= 1.582

START:Continue; Select function.:＿

□1 Remeasure

Fig. 3-46 Example of Display after 1st-round

Sample Measurement

(4) When the specified number of samples have all been

measured, the screen shown in Fig. 3-47 appears.
For the processing function assigned to each key, refer
to Table 3-34.

3 - 76
 3.2.3

Photometry CELL A
ID ABS
10 WL1 （ 260.0nm） 0.423
WL2 （ 280.0nm） 0.268

ABS DIFF= 0.155

ABS Ratio= 1.582

START:Continue; Select function.: ＿

□1 Meas. □
2 Save □3 Print

Fig. 3-47 Example of Display after Sample

Measurement
(number of samples: 10)

Table 3-34 Guidance after Sample Measurement

Setting Item Description

[1] Meas. You can make setting related to measurement.
“New Meas.”, “Continue” and “Remeasure” are
selectable.
[1] New Meas. : Selected when newly measuring
samples with the same calibration
curve.
[2] Continue : Allows you to start measurement
from the number following the
previously measured sample.
When this function is selected, the
guidance given below appears.
Enter the number of samples to be
measured and press the [ENTER]
key.

Input value. ： ＿
No.of Sample [1-140]

Display with 10 samples measured

[3] Remeasure : Selected when remeasuring a

measured sample. When this
function is selected, the guidance
given below appears. Select a
sample to be remeasured with the
[▲] or [▼] key, set the sample to
cell 1 and press the [START] key to
carry out remeasurement.
To execute auto zero, press the
[AUTOZERO] key.

3 - 77
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
Setting Item Description
[1] Meas. Photometry CELL A
ID ABS
10 WL1 （ 260.0nm） 0.423
WL2 （ 280.0nm） 0.268

ABS DIFF= 0.155

ABS Ratio= 1.582

Set sample to Cell1(Sample10).: _＿

START:Meas.; AUTOZERO:Autozero
W X( 10/ 10)

[2] Save You can save the measured data. When this
function is selected, the following screen appears.

Save Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
□1 Save □
2 Sort
W X ( 1/ 1)

[1] Save : Allows you to save data under a new file

name.
[2] Sort : Allows you to rearrange the currently
saved data in the order of file names or
dates.
[3] Print You can select items to be printed. When this
function is selected, the screen given below appears.
For each item, select whether to print out (ON or
OFF).

Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Sample ON

Select function. : _
□1 ON □
2 OFF

3 - 78
 3.2.4

3.2.4 Measuring a Spectrum

This instrument is capable of measuring the transmission spectrum and absorption spectrum.
By setting the blank to cell A, one sample can be measured after automatic execution of
baseline correction.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Setting of Measuring Conditions

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 3-48) will appear. To set each condition for
wavelength scan, press the <2> key (WL Scan) and
then [ENTER] key. Or select “WL Scan” with the [▲]
or [▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 3-48 Main Menu Screen

(2) The WL Scan screen (Fig. 3-49) appears. To set the

measuring conditions, press the <1> key (Meas.
PARAM.) and then [ENTER] key. Or select “Meas.
PARAM.” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

3 - 79
3.2 Automatic Continuous Measurement (6-cell auto mode)

WL Scan CELL A
1 Meas.PARAM. Data Mode ABS
2 Sample Start WL(nm) 600.0
3 6 Cell End WL(nm) 400.0
4 System Upper Scale 1.000
5 Peak PARAM. Lower Scale 0.000
6 Print Scan Speed 400
7 Save Method Delay(s) 0
8 Meas.Screen Data Int. Normal

Select function. : _

Fig. 3-49 WL Scan Screen

(3) The measuring condition (Meas. PARAM.) screen (Fig.

3-50) appears. With the [▲] or [▼] key, set the data
mode (Data Mode), start wavelength (Start WL), end
wavelength (End WL), scan speed (Speed), initial
delay (Delay), Y-axis upper limit (Upper Scale) and Y-
axis lower limit (Lower Scale) according to the
guidance. For details of each parameter, refer to
Table 3-35. After completion of setting, return to the
WL Scan screen by pressing the [RETURN] key or [◄]
key.

WL Scan CELL A
1 Meas.PARAM. Data Mode ABS
2 Sample StartWL(nm) 600.0
3 6ｾﾙ End WL(nm) 400.0
4 System Upper Scale 1.000
5 Peak PARAM. LowerScale 0.000
6 Print Scan Speed 400
7 Save Method Delay(s) 0
8 Meas.Screen Data Int. Normal

Select function. : _
1 ABS □
□ 2 %T

Fig. 3-50 Measuring Condition (Meas. PARAM.)

Screen

3 - 80
 3.2.4

Table 3-35 “Meas. PARAM.” Setting Parameters

Setting Item Description

Data Mode Determine the unit of scale on the Y axis.
[1] ABS : Used for measuring the absorption
spectrum (spectrum with absorbance on
the Y axis).
[2] %T : Used for measuring the transmission
spectrum (spectrum with transmittance on
the Y axis).
Start WL Enter a wavelength for starting wavelength scanning.
(nm) This parameter is specifiable in a range of 200.0 to
1100.0 nm in steps of 0.1 nm.
Make your setting so that Start WL < End WL.
End WL (nm) Enter a wavelength for ending wavelength scanning.
This parameter is specifiable in a range of 190.0 to
1090.0 nm in steps of 0.1 nm.
Make your setting so that Start WL < End WL.
Upper Scale Enter a Y-axis upper limit for the spectrum displayed
during measurement.
This parameter is specifiable in the following range.
ABS : -9.999 to 9.999
%T : -999.9 to 999.9
Lower Scale Enter a Y-axis lower limit for the spectrum displayed
during measurement.
This parameter is specifiable in the following range.
ABS : -9.999 to 9.999
%T : -999.9 to 999.9
Scan Speed Set up a wavelength scanning speed. The following
7 speeds are selectable.
[1] 40 [2] 100 [3] 200 [4] 400 [5] 800 [6] 1200
[7] 2400
The data interval varies with the scan speed. For
this interval, a small value is set at low speed and a
large value at high speed. For details, refer to the
description of “Data Int.”
Delay (s) After the [START] key is pressed, the system waits
for the time period set here and then starts
measurement. This parameter is settable in a range
of 0 to 9999 sec in steps of 1 sec.
Set the parameter when you want to start
measurement after a specified time period, including
when measuring a sample after its temperature
reaches the room temperature and when starting
measurement after completion of reaction. Enter “0”
if the delay time need not be set.

3 - 81
3.2 Automatic Continuous Measurement (6-cell auto mode)

(cont’d)
Setting Item Description
Data Int. Set up a data interval. “Normal” and “Fine” are
selectable.
[1] Normal : Usually selected.
[2] Fine : Permits measurement at short data
intervals as compared with Normal.
However, noise is more likely to be
generated on a spectrum, because the
data acquisition time per point is
shorter than when Normal is selected.
The actual data interval differs
depending on the scan speed and
measuring wavelength range.
Whichever is longer is made effective.
In case of scanning at 400 nm/min in a
range of 600 to 400 nm for example,
the data interval is set to 2 nm when
Normal is selected, and to 1 nm when
Fine is selected.

Scan speed and data interval

Scan Speed Data Sampling Interval (nm)
(nm/min) Normal Fine
40 0.2 0.1
100 0.5 0.2
200 1 0.5
400 2 1
800 3 2
1200 5 3
2400 5 5

Data interval according to measuring wavelength

range
Measuring wavelength Data
range (nm) interval (nm)
λ > 500 1.0
500 ≥ λ > 200 0.5
200 ≥ λ > 100 0.2
100 ≥ λ 0.1

3. Setting of Sample Conditions

(1) To set the sample conditions, press the <2> key

(Sample) and then [ENTER] key. Or select “Sample”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

3 - 82
 3.2.4

(2) The sample condition (Sample) screen (Fig. 3-51)

appears. Enter a sample name here. For details,
refer to Table 3-36. After input, return to the WL Scan
screen by pressing the [ENTER] key or [◄] key.

WL Scan CELL A
^1 Meas.PARAM. Sample Name
2 Sample
3 6 Cell
4 System
5 Peak PARAM.
6 Print
^7 Save Method
8 Meas.Screen

Input character. ： ＿
Sample Name [Max: 8 characters]

Fig. 3-51 Sample Condition (Sample) Screen

Table 3-36 Input of Sample Name

Setting Item Description

Sample Up to 8 half-size alphanumeric characters can be
Name entered for the sample name. The sample name
entered here is printed in the sample name field on
the report. For the character input procedure, refer
to section 2.4.3.

4. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <3> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

3 - 83
3.2 Automatic Continuous Measurement (6-cell auto mode)

(2) The 6-cell condition (6 Cell) screen (Fig. 3-52) appears.

To set each item on the 6-cell condition (6 Cell) screen,
press the [ENTER] key. Or select each item with the
[▲] or [▼] key and make your setting according to the
guidance. For details of each parameter, refer to
Table 3-37.

WL Scan CELL A
^1 Meas.PARAM. 6 Cell Mode Auto
2 Sample No.of Sample 1
3 6 Cell
4 System
5 Peak PARAM.
6 Print
^7 Save Method
8 Meas.Screen

Select function. : _
□1 Auto □
2 Manual

Fig. 3-52 6-cell Condition (6 Cell) Screen

Table 3-37 “6 Cell” Setting Parameters

Setting Item Description

6 Cell Mode Select a 6-cell operation mode for measurement from
the following.
[1] Auto
By setting a baseline sample and unknown samples
to the 6-cell holder, it is possible to automatically turn
the 6-cell holder and automatically measure one
sample after baseline correction.
[2] Manual
Baseline correction and sample measurement are
performed manually. It is required to manually move
the cell and execute baseline correction.
(For details of the Manual mode, refer to section
3.3.4..)
No. of Set the number of samples to be measured.
Sample This parameter is settable in a range of 1 to 5.

(3) After setting each item, return to the WL Scan screen by

pressing the [RETURN] key or [◄] key.

5. Setting of System Conditions

(1) To set the system conditions, press the <4> key

(System) and then [ENTER] key. Or select “System”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

3 - 84
 3.2.4

(2) The system condition (System) setting screen (Fig. 3-

53) appears. Set a response according to the
guidance. For detail, refer to Table 3-38.

WL Scan CELL A
^1 Meas.PARAM. Response Medium
2 Sample
3 6 Cell
4 System
5 Peak PARAM.
6 Print
^7 Save Method
8 Meas.Screen

Select function. : _
□1 Fast □
2 Medium □
3 Slow

Fig. 3-53 System Condition (System) Setting

Screen

Table 3-38 Response Setting Parameter

Setting Item Description

Response Select from the following 3 response speeds.
[1] Fast : Used for high-resolution measurement
with respect to the wavelength. Note
that the noise level is high as
compared with Medium and Slow.
[2] Medium: Used for ordinary measurement.
[3] Slow : Used to reduce a variation in
photometric value. This response
speed is unsuitable for high-resolution
measurement as compared with Fast
and Medium.

(3) After completion of response setting, return to the WL

Scan screen by pressing the [RETURN] key or [◄] key.

6. Setting of Peak Detection Conditions

(1) To set the peak detection conditions, press the <5>

key (Peak PARAM.) and then [ENTER] key.
Or select “Peak PARAM.” with the [▲] or [▼] key and
press the [ENTER] or [►] key.

(2) The peak detection condition (Peak PARAM.) setting

screen (Fig. 3-54) appears. Specify a threshold and
sensitivity here according to the guidance. For details,
refer to Table 3-39.

3 - 85
3.2 Automatic Continuous Measurement (6-cell auto mode)

WL Scan CELL A
^1 Meas.PARAM. Threshold 0.010
2 Sample Sensitivity 2
3 6 Cell
4 System
5 Peak PARAM.
6 Print
^7 Save Method
8 Meas.Screen

Input value. ： ＿
Threshold[0.001-1.000]

Fig. 3-54 Peak Detection Condition (Peak

PARAM.) Setting Screen

Table 3-39 “Peak PARAM.” Setting Parameters

Setting Item Description

Threshold Set up a condition for detecting a peak and valley
from the measured spectrum. In peak detection,
peaks including noise are detected if the threshold
level is reduced. Conversely, small peaks cannot be
detected if the threshold level is increased. As just
described, it is a threshold level that determines the
detectability in the Y-axis direction of a spectrum.
If the minimum difference in photometric value
between the peak and valley (peak-valley difference
shown in the following figure) is larger than the
threshold level, they are detected as a peak and
valley.

Detected peak and valley: Minimum peak-valley

difference > Threshold level

This parameter is specifiable in the following range.

ABS : 0.001 to 1.000
valley difference

%T : 0.1 to 100.0
Minimum peak-

Peak
Absorbance or
transmittance

Valley

Wavelength

3 - 86
 3.2.4

(cont’d)
Setting Item Description
Sensitivity Set up a condition for detecting a peak and valley
from the measured spectrum.
It is sensitivity that determines the detectability in the
X-axis direction of a spectrum. For detection of a
sharp peak, select sensitivity 1. For detection of a
broad peak, select sensitivity 8. The following 4
levels are selectable.
[1] 1
[2] 2
[3] 4
[4] 8
For details, refer to Explanation 3-9.

(3) After completion of calculation condition setting, return

to the WL Scan screen by pressing the [RETURN] or
[◄] key.

3 - 87
3.2 Automatic Continuous Measurement (6-cell auto mode)

Explanation 3-9 Peak Detection Sensitivity

The setting of peak detection sensitivity is employed for judging whether data has increased or
decreased by comparing the data at the present base point and the data after N points, based on
which a peak will be detected. Explanation is given here in the case of peak detection where peak
sensitivity is set at level 1 (data after 6 points to be used for comparison).

When the data after N points is larger than at the present base point, it is taken as a rise.
When smaller, it is taken as a fall. Thus, data at the next base point is compared with that after N
points. This sequence is repeated for rise or fall judgments. (See Fig. 1.) When a rise occurs
N/2 times in succession followed by N/2-times succession of a fall, it is judged that a peak is
detected. Also, when a fall occurs N/2 times in succession followed by N/2-times succession of a
rise, it is judged that a valley is detected. (See Fig. 2.)

D5 D6
D4 D7
D3 D8
D2 D9
D1 D10
D0 D11

Fall
Fall
Fall
Rise
Rise
Rise

N points
Fig. 1 Schematic Diagram of Peak

Table 1 Rule on Rise or Fall Judgment

Base-point data D0 D1 D2 D3 D4 D5
Data after N points D6 D7 D8 D9 D10 D11
Rise/fall judgment Rise Rise Rise Fall Fall Fall

Sensitivity setting assigns a concrete number to “N” which represents the number of points after the
base point to be used for peak detection. Table 2 lists the relation between sensitivity level and
data checking interval. At sensitivity level 1, data after 6 points is checked. At sensitivity level 8,
data after 48 points is checked. Therefore, sensitivity level 1 should be selected for detecting
peak/valley in a sharp spectrum, and sensitivity level 8 for detecting peak/valley in a spectrum
having a moderate curve.

Table 2 Sensitivity Level and Data Checking Interval

Sensitivity Level Interval (N points)

1 6 points
2 12 points
4 24 points
8 48 points

3 - 88
 3.2.4

7. Setting of Printing Conditions

GUIDE: Specify print items if determined beforehand.

You can also specify print items after completion
of measurement. If you do not want to set the
printing conditions here, proceed to step 8.

(1) To set the printing conditions, press the <6> key (Print)
and then [ENTER] key. Or select “Print” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The printing condition (Print) screen (Fig. 3-55)

appears. Make your setting according to the
guidance. For details of each parameter, refer to
Table 3-40.

WL Scan CELL A
1 Meas.PARAM. Auto Print OFF
2 Sample Print Date ON
3 6 Cell Run Date ON
4 System Method ON
5 Peak PARAM. Graph ON
6 Print Peak Table ON
7 Save Method Data List OFF
8 Meas.Screen

Select function. : _
□1 ON □2 OFF

Fig. 3-55 Printing Condition (Print) Screen

(3) After completion of setting for Auto Print, Print Date,

Run Date, Method, Graph, Peak Table, Data List and
Int., return to the WL Scan screen by pressing the
[RETURN] key or [◄] key.

3 - 89
3.2 Automatic Continuous Measurement (6-cell auto mode)

Table 3-40 “Print” Setting Parameters

Location in
Setting Item Description
Fig. 3-56
Auto Print [1] ON : Automatic printing is carried ⎯
out after measurement.
[2] OFF : Automatic printing is not
carried out after
measurement.
Print Date [1] ON : Printing date is printed. <1>
[2] OFF : Printing date is not printed.
Run Date [1] ON : Analysis date is printed. <2>
[2] OFF : Analysis date is not printed.
Method [1] ON : Measuring conditions are <3>
printed.
[2] OFF : Measuring conditions are
not printed.
Graph [1] ON : Spectrum is printed. <4>
[2] OFF : Spectrum is not printed.
Peak Table [1] ON : Peak table is printed. <5>
[2] OFF : Peak table is not printed.
Data List [1] OFF : Numerical data of <6>
spectrum is not printed.
[2] All Data : Numerical data of
spectrum is all printed.
[3] Interval : Numerical data of
spectrum is printed at
the specified intervals.
Int. Specify the printing interval for the data ⎯
list. This parameter is indicated when
“Interval” is selected for Data List. It
is specifiable in a range of 0.5 to 100.0.

3 - 90
 3.2.4

Report：2010/04/10 15:12 ……….. <1>

0.800

<4>

0.000
400.0 462.5 525.0 587.5 650.0 nm

Sample Name： ……….. <2>

File Name：
Run Date： 2010/04/10 13:34
Operator：

Spectrophotometer
Model： U-5100 Spectrophotometer
Serial No.： 1234-567
ROM VER.： 3J25300
Option：

Instrument
Meas.Mode： WL Scan Response： Medium
Data Mode： ABS Bandpass(nm)： 5.0
Start WL(nm)：
End WL(nm)：
650.0
400.0
Baseline：
6 Cell Mode：
User
Auto
<3>
Speed(nm/min)： 200 No.of Sample： 1
Delay(s)： 0
Data Int.： Fine

Peak Table(Threshold:0.010 Sene:2)

No. WL(nm) Peak WL(nm) Valley
1 542.5 0.650 <5>

Data List(Int.(nm): 1.0)

No. WL(nm) ABS No. WL(nm) ABS No. WL(nm) ABS
1 650.0 0.000 3 648.0 0.001 5 646.0 0.002
2 649.0 0.000 4 647.0 0.001 6 645.0 0.002 <6>

Fig. 3-56 Example of Printout in WL Scan Mode

8. Saving of Measuring Conditions

GUIDE: When the set measuring conditions need not be

saved, proceed to step 9.

(1) To save the set measuring conditions, press the <7>

key (Save Method) and then [ENTER] key. Or select
“Save Method” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(2) The measuring condition saving (Save Method) screen

(Fig. 3-57) appears.

3 - 91
3.2 Automatic Continuous Measurement (6-cell auto mode)

Meas.PARAM. CELL A
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

File name Measurement Saving

mode date

File Name：＿
File Name [Max: 20 characters]
ABC W X ( 1/ 1)

Fig. 3-57 Measuring Condition Saving (Save

Method) Screen

(3) When other measuring conditions are saved in

advance, a list of saved measuring conditions appears
on the main menu. At this time, the file name,
measurement mode (PHT: Photometry, WLS:
Wavelength Scan, TMS: Time Scan) and saving date
are indicated.

(4) For the measuring conditions to be saved, enter a file

name within 20 characters and press the [ENTER] key.

NOTE:
• If a file of the same name as entered already exists in
the same measurement mode, the following guidance
appears.
Already exists. Overwrite it? :_
1 Yes
□ 2 No
□

For overwriting, press the <1> key (Yes). To avoid

overwriting, press the <2> key (No) and rename the file
to be saved.
• A total of 50 method files can be saved for all
measurement modes.
• If 50 files are already registered, the following message
appears.
No.of files is full.

In this case, delete files referring to section 4.2.2 and

retry saving.
• The method for which auto start (for details, refer to
section 4.5.) is set is preceded by “*”.

3 - 92
 3.2.4

(5) Return to the WL Scan screen by pressing the

[RETURN] key.

9. Setting and Measurement of Samples

(1) To determine the measuring conditions set in the

preceding steps, press the <8> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key.

(2) The guidance on sample setting (Fig. 3-30) appears.

On the screen, displayed from the left are the cell
number and sample name. Cell A is used for baseline
correction. Set a baseline correction sample (blank)
to cell A. And set the corresponding samples to cells
1 to 5. Upon completion of setting, press the [START]
key. After baseline correction with cell A, sample 1
will be measured.

WL Scan CELL A
[Info]
Set samples to Cell No..
Cell A Blank
Cell 1 Sample1
Cell 2 Sample2
Cell 3 Sample3
Cell 4 -
Cell 5 -

START:Meas.Baseline and Sample1.: _

Fig. 3-58 Guidance on Sample Setting

10. Measurement of Sample Solutions

(1) Upon completion of sample 1 measurement, the

guidance shown in Fig. 3-59 appears. To record data,
perform the save or print operation. To save data,
select the <3> key (Save). To print data, press the
[PRINT] key. Pressing the <1> to <5> keys allows the
processings shown in Table 3-41. For proceeding to
the next measurement, press the [START] key.

3 - 93
3.2 Automatic Continuous Measurement (6-cell auto mode)

NOTE: In WL Scan mode, the spectrum of each sample

cannot be automatically saved. So the save or
print operation is required for each sample.

WL Scan CELL A

0.800

ABS

0.000
400.0 525.0 650.0nm
START:Meas.; Select function.：_
□1 Data □
2 Scale □
3 Save □
4 Print □
5 Baseline

Fig. 3-59 Screen after Sample Measurement

Table 3-41 Guidance after Measurement in WL Scan Mode

Setting Item Description

[1] Data You can perform each data processing. When this
function is selected, the following guidance appears.

Select function. : _
1 Trace
□ 2 Peak □
□ 3 Smoothing □
4 List

For details of the function, refer to section 4.3.

[1] Trace : Allows you to read out all the
measured data on the spectrum.
[2] Peak : Displays the detected peak.
[3] Smoothing : Allows you to perform spectrum
smoothing.
[4] List : Allows you to display the measured
data at the specified intervals.
[2] Scale You can change the scale on the X axis and Y axis of
spectrum.
For detail of this function, refer to section 4.3.

3 - 94
 3.2.4

(cont’d)
Setting Item Description
[3] Save You can save the measured spectrum. When this
function is selected, the following screen appears.

Save Data CELL A

1 NO3-(Hitachi Taro) WLS 2010/10/10
2 Cr6+(Hitachi Hanako) WLS 2010/03/30

Select function. : _
□1 Save □
2 Sort
W X ( 1/ 1)

[1] Save : Allows you to save data under a new file

name.
[2] Sort : Allows you to rearrange the currently
saved data in the order of file names or
dates.
[4] Print You can select items to be printed. When this
function is selected, the screen given below appears.
For each item, select whether to print out (ON or
OFF).

Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Graph ON
Peak Table ON
Data List ON

Select function. : _
□1 ON □
2 OFF
W X ( 1/ 1)

[5] Baseline You can execute baseline correction. When this

function is selected, the following guidance appears.

Select FUNC. and execute at Cell A.:＿

1 Yes
□ 2 No
□

[1] Yes : Selected to execute baseline correction.

Set a sample for baseline correction to cell
A and press the <1> key.
[2] No : Selected to return to the previous screen
without executing baseline correction.

3 - 95
3.2 Automatic Continuous Measurement (6-cell auto mode)

(2) Before proceeding to the next measurement, the

system displays the data deletion confirming screen
(Fig. 3-60). When data has already been saved or
printed, select the <1> key (Yes). If data has not yet
been saved or printed, select the <2> key (No) and
perform the save or print operation.

Data will be deleted. Continue? :＿

1 Yes
□ 2 No
□

Fig. 3-60 Data Deletion Confirming Screen

(3) Pressing the [START] key starts spectrum

measurement. After measurement, you can execute
the same operation as in (1). By repeating this
operation, it is possible to perform spectrum
measurement for up to 5 samples.

START:Meas.Sample2：_＿

Fig. 3-61 Guidance on Measurement Start

3 - 96
 3.3

3.3 Sample-by-sample Measurement (6-cell manual mode)

Samples are measured one by one. This mode is suitable for

measurement of multiple samples with 2 cells (one for auto zero).
By pressing the [Autozero] key, it is possible to correct the
absorbance to zero with cell A set in advance.

3
4 2

5 1
A
1

Measurement of solution concentration →section3.3.1.

Generation of calibration curve and determination of
unknown sample concentration →section 3.3.1.
Determination of unknown sample concentration
by entered calibration curve factor →section 3.3.1.
Determination of unknown sample concentration
from saved calibration curve →section 4.1.4.
Measurement of absorbance/transmittance →section 3.3.2.
Measurement of DNA →section 3.3.3.
Measurement of spectrum →section 3.3.4.
Measurement of change with time →section 3.3.5.

3 - 97
3.3 Sample-by-sample Measurement (6-cell manual mode)

3.3.1 Determining Concentration of Solution

This instrument is used for generating a calibration curve and determining the concentration
of an unknown sample, and for entering a calibration factor and determining the
concentration.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Setting of Measuring Conditions

3. Setting of Sample Conditions

*
4. Setting of Calibration Curve Conditions

5. Setting of Calibration Curve Data

* For steps 2 to 5, refer to section 3.2.1..

6. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <5> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

3 - 98
 3.3.1

(2) The 6-cell condition (6 Cell) screen (Fig. 3-62)

appears.

Photometry CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample STD Autozero STD1
3 Curve SAMP.Autozero ON
4 Curve Data Autozero int. 5
5 6 Cell No.of Sample 1
6 System
7 Print
8 Save Method
9 Meas.Screen
Select function. : _
1 Auto □
□ 2 Manual

Fig. 3-62 6-cell Condition (6 Cell) Screen

(3) To set each item on the 6-cell condition (6 Cell)

screen, press the [ENTER] key. Or select each item
with the [▲] or [▼] key and make your setting
according to the guidance. For details of each
parameter, refer to Table 3-42.

Table 3-42 “6 Cell” Setting Parameter

Setting Item Description

6 Cell Mode Specify a 6-cell operation mode for
measurement.
[1] Auto
In this mode, a sample is set at each
position of the 6-cell holder beforehand and
the holder is automatically turned to
automatically perform from auto zero to
standard/sample measurement. It is
recommended to use this mode if you have
6 cells and there are many samples to be
measured.
(For details, refer to section 3.2.1..)
[2] Manual
In this mode, standard/sample
measurement is carried out by using one
position (cell 1 to 5 selectable) of the 6-cell
holder. To eliminate the influence of
instrument drift (baseline fluctuation) during
measurement, auto zero can easily be
executed by setting a solution for auto zero
to cell A. In normal usage, set a standard
or sample solution to cell 1.

3 - 99
3.3 Sample-by-sample Measurement (6-cell manual mode)

7. Setting of System Conditions

*
8. Setting of Printing Conditions

* For steps 7 and 8, refer to section 3.2.1..

9. Measurement of Standard Solutions

GUIDE: When 2) Factor is selected for Curve Type in

step 4, standard solution measurement is not
carried out. So proceed to step 10.

(1) To determine the measuring conditions set in the

preceding steps, press the <9> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key.

(2) The auto zero sample setting screen (Fig. 3-63)

appears. Set an auto zero sample to cell A.
The lamp is lit for 30 seconds and the current
measured value is indicated so that you can check if
execution of auto zero is necessary.
Upon completion of setting, press the [AUTOZERO]
key. Auto zero will be executed at the position of cell
A. When avoiding auto zero, press the [CLEAR] key
to cancel auto zero.

NOTE: For the first measurement under the set

conditions, be sure to execute auto zero.

For details of the auto zero sample, refer to

Explanation 3-10 and Explanation 3-11.

Photometry 600.0 nm 0.000 ABS CELL A

[Info]
Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Fig. 3-63 Auto Zero Sample Setting Screen

3 - 100
 3.3.1

Explanation 3-10 Auto Zero Method (without coloring reagent)

Standard solution Sample solution

Absorbance of
Absorbance

Analyte of interest analyte

Solvent Absorbance-zero line by auto

zero with <1>
Cell
<1> <2> <3> <4>
STD1 STD2 STD3 Sample

Elements causing absorbance without using coloring reagent (not containing interfering substance)

A measured absorbance value is given as a total of absorptions due to various elements (cell,
solvent, coloring reagent, analyte of interest, interfering substance). In other words, this value
does not simply stand for an absorption due to the analyte alone.
For generating a calibration curve according to concentration levels 0, 1 and 2, solutions of
concentration levels 0, 1 and 2 are prepared for STD1, STD2 and STD3, respectively (see <1> to
<3> in the above figure). These STD’s have absorptions due to cell, solvent and analyte.
However, the absorbance required for actual quantitation is the one deriving from the analyte.
Therefore, absorptions due to cell and solvent need to be subtracted (by auto zero operation).
In this case, auto zero is carried out with STD 1 and the absorbance values of STD1 to STD3 are
measured, thereby generating a calibration curve.
And, for sample quantitation, the concentration of the sample is determined using the absorbance
after exclusion of cell and solvent. When the absorption other than those due to cell and solvent
derives from the analyte alone, the determined concentration equals the concentration of the
analyte as illustrated at <4> in the above figure.

Standard solution Sample solution

Absorbance of
Interfering analyte
Absorbance

Analyte of interest
substance
Absorbance-zero line by auto
zero with <5>
Absorbance-zero line by auto
Solvent zero with <1>
Cell
<1> <2> <3> <5> <6>
STD1 STD2 STD3 Blank test Sample (interfering
solution substance contained)

Elements causing absorbance without using coloring reagent (containing interfering substance)

In the case where a sample contains an interfering substance such as turbid or disturbing
component as illustrated at <6> in the above figure, the absorbance due to the interfering substance
will be added to the quantitative value of the analyte.
In this case, it is necessary to carry out a blank test (the same preparatory operation as for samples
by use of pure water or the like) and subtract the measured absorbance of the blank test solution by
auto-zero operation at the time of sample measurement or subtract the quantitative value of the
blank test solution from that of the sample in order to subtract the absorbance of the interfering
substance from that of the sample.

3 - 101
3.3 Sample-by-sample Measurement (6-cell manual mode)

Explanation 3-11 Auto Zero Method (with coloring reagent)

Standard solution Sample solution

Absorbance of
Analyte of interest analyte
Absorbance

Coloring
reagent
Absorbance-zero line by auto
zero with <1>
Solvent
Cell
<0> <1> <2> <3> <4>
Blank (solvent STD1 STD2 STD3 Sample
and cell)

Elements causing absorbance with coloring reagent (not containing interfering substance)

When using a coloring reagent, the absorption of a sample solution includes that of this reagent
besides those of cell and solvent. For subtracting the absorbance due to the coloring reagent as
well, auto zero is carried out with the solution at <1> in the above figure.
For sample quantitation, the concentration of the sample is determined using the absorbance after
exclusion of cell, solvent and coloring reagent. When the absorption other than those due to cell,
solvent and coloring reagent derives from the analyte alone, the determined concentration equals the
concentration of the analyte as illustrated at <4> in the above figure.
Also, when the absorbance of the solution at <1> in the above figure is unstable with time, the solution
is not suited for auto zero operation. Therefore, it is recommended to carry out auto zero operation
with the blank (solvent and cell).

Standard solution Sample solution

Absorbance of analyte
Analyte of Interfering
interest substance Absorbance-zero line by auto
Absorbance

zero with <5>

Coloring Absorbance-zero line by auto

reagent zero with <1>
Solvent
Cell
<0> <1> <2> <3> <5> <6>
Blank (solvent STD1 STD2 STD3 Blank test Sample (interfering
and cell) solution substance contained)

In the case where a sample contains an interfering substance such as turbid or disturbing component
as illustrated at <6> in the above figure, the absorbance due to the interfering substance will be added
to the quantitative value of the analyte. In this case, it is necessary to carry out a blank test (the
same preparatory operation as for samples by use of pure water or the like) and subtract the
measured absorbance of the blank test solution by auto-zero operation at the time of sample
measurement or subtract the quantitative value of the blank test solution from that of the sample in
order to subtract the absorbance of the interfering substance from that of the sample.

3 - 102
 3.3.1

(3) The standard measurement screen (Fig. 3-64) appears.

At this time, the cell position shifts to cell 1.
For measurement at a different position, hold down the
[SHIFT] key and press the [◄] or [►] to shift the cell
position. Make sure that the aimed-at cell number is
indicated on the screen.

NOTE: In 6-cell manual mode, the photometric value at

the cell position indicated on the screen is
measured. Before measurement, therefore,
check if the cell position where the standard has
been set coincides with the cell position
indicated on the screen.

GUIDE: In 6-cell manual mode, the photometric value at

the cell position indicated on the screen is
measured. For carrying out auto zero
frequently, it is recommended to use cell 1.
Cell 1 can move to the position of cell A in a short
time, thus ensuring smooth measurement.
When the guidance is displayed, auto zero can
be executed at the position of cell A by pressing
the [AUTOZERO] key. After execution of auto
zero, the system moves to the cell position before
the execution. By pressing the [AUTOZERO]
with the auto zero sample set to cell A and the
sample set to cell 1, you can execute auto zero
without taking the cell in and out.

Cell
position
Photometry CELL 1

ID ABS CONC
STD1 0.000
STD2 10.00
STD3 20.00

START:Meas.STD1：＿

Example of display with “3” set for

number of standards

Fig. 3-64 Standard Measurement Screen

3 - 103
3.3 Sample-by-sample Measurement (6-cell manual mode)

Figure 3-65 shows the standard setting screen.

On the screen, displayed from the left are the standard
name, measured absorbance and standard
concentration. Measurement is carried out
sequentially starting from STD1, until the specified
number of standards are all measured. According to
the guidance, set the indicated standard to the cell
position indicated on the screen (cell 1), close the
sample compartment cover and press the [START] key.
Standard name Photometry CELL 1

ID ABS CONC
STD1 0.000 Standard
STD2 10.00 concentration
STD3 20.00

Measured absorbance
START:Meas.STD1：＿

Example of display with “3” set for

number of standards

Fig. 3-65 Standard Setting Screen

(4) During measurement, the screen shown in Fig. 3-66 is

displayed.

Photometry 600.0 nm 0.001 ABS CELL 1

ID ABS CONC
STD1 0.000
STD2 10.00
STD3 20.00

Meas.STD1...

Example of display with “3” set for

number of standards

Fig. 3-66 Screen Displayed during Standard

Measurement

3 - 104
 3.3.1

(5) Upon completion of measurement, the absorbance of

STD1 is indicated on the screen (Fig. 3-67).
This screen appears after completion of measurement
of each standard. Pressing the [START] key here
starts measurement of the next standard (STD2).
Pressing the <1> key (Curve Confirmation) or <2> key
(Remeasure) allows the processing shown in Table 3-
43.

Photometry CELL 1

ID ABS CONC
STD1 0.001 0.000
STD2 10.00
STD3 20.00

START:Meas.STD 2; Select FUNC..：＿

1 Curve Confirmation □
□ 2 Remeasure

Example of display with “3” set for

number of standards

Fig. 3-67 Screen after Standard Measurement

Table 3-43 Guidance during Standard Measurement

Setting Item Description

[1] Curve You can generate a calibration curve with
Confirmation the standards measured before now and
confirm it.
Photometry/Curve CELL 1
R = 0.9999, R2 = 0.9999
0.405 +
+
ABS +
+
0.001 +
0.000 CONC 40.000
Select function. : _
1 Scale □
□ 2 Factor □ 3 STD

[1] Scale :
Allows you to change the
scale of calibration curve.
[2] Factor : Indicates the factor,
correlation coefficient and
determination coefficient of
calibration curve.
[3] STD : Indicates the result of
standard solution
measurement.
[2] Remeasure You can remeasure the last-measured
standard.

3 - 105
3.3 Sample-by-sample Measurement (6-cell manual mode)

(6) When the specified number of standards have all been

measured, the screen shown in Fig. 3-68 appears.
When the <1> key (Sample Meas.) is pressed, the
system terminates standard measurement and
proceeds to sample measurement. Pressing the <2>
key (Curve) or <3> key (Save) allows the processing
shown in Table 3-44.

Photometry CELL 1

ID ABS CONC
STD1 0.001 0.000
STD2 0.102 10.00
STD3 0.205 20.00

Select function. : _
1 Sample Meas. □
□ 2 Curve □
3 Save □
4 Print

Fig. 3-68 Screen after Measurement of All Standards

3 - 106
 3.3.1

Table 3-44 Guidance after Standard Measurement

Setting Item Description

[1] Sample Meas. The system proceeds to sample
measurement.
[2] Curve You can generate a calibration curve with
the measured standards and confirm it.
When this function is selected, the
following screen appears.
Photometry/Curve CELL A
R = 0.9999, R2 = 0.9999
+
0.893 +
+
+
ABS +
+
+
+
0.001 +
0.000 CONC 90.000
Select function. : _
1 Scale □
□ 2 Factor □ 3 STD

[1] Scale :
Allows you to change the
scale of calibration curve.
[2] Factor : Indicates the factor,
correlation coefficient and
determination coefficient of
calibration curve.
[3] STD : Indicates the result of
standard solution
measurement. Also allows
remeasurement of a
standard solution, deletion
of standard data
unnecessary for generation
of a calibration curve or
retrieval of deleted data.
[3] Save You can save the measured standard
data. When this function is selected, the
following screen appears.
Save Data CELL A
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
1 Save □
□ 2 Sort
◄ ► ( 1/ 1)

[1] Save : Allows you to save the result

data under a new file name.
[2] Sort : Allows you to rearrange the
presently saved data in the
order of file names or dates.

3 - 107
3.3 Sample-by-sample Measurement (6-cell manual mode)

(cont’d)
Setting Item Description
[4] Print You can select items to be printed.
When this function is selected, the screen
given below appears. For each item,
specify whether to print out (ON or OFF).
Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Curve ON
STD ON
Sample ON

Select function. : _
1 ON □
□ 2 OFF

10. Measurement of Sample Solutions

(1) The sample measurement screen (Fig. 3-69) appears.

Set the sample indicated in the guidance area in the
sample compartment. Pressing the [START] key
starts measurement at the current cell position.

NOTE: In 6-cell manual mode, the photometric value at

the cell position indicated on the screen is
measured. Before measurement, therefore,
check if the cell position where the sample has
been set coincides with the cell position
indicated on the screen.

Photometry CELL 1
ID ABS CONC
1

START:Meas.SAMP.1; Select FUNC..：＿

1 Curve Confirmation
□

Fig. 3-69 Sample Measurement Screen

3 - 108
 3.3.1

Auto zero can be executed by pressing the

[AUTOZERO] key when the guidance on sample
setting is indicated. For details, refer to Fig. 3-70.

<1> Check that a guide message “START: Meas.

SampleXX:_” is displayed. For carrying out auto
zero, auto zero sample should be set to cell A.
Photometry CELL 1

ID ABS CONC
1

START:Meas.Sample1：＿

Sample Measurement Screen

<2> Set an auto zero sample, close the cover and press
the [AUTOZERO] key. The 6-cell holder will move to
the position of cell A and auto zero will be executed
there
<3> After execution of auto zero, the 6-cell holder moves
to the position before the execution and the sample
measurement screen reappears.
GUIDE: For executing auto zero frequently, set an auto zero
to the position of cell A beforehand. It is now
possible to move to the position of cell A and
execute auto zero just by pressing the [AUTOZERO]
key.

Fig. 3-70 How to Execute Auto Zero

(2) After measurement of a sample, its measured

absorbance and concentration are indicated at the
corresponding sample position on the screen.
This screen appears after measurement of each
sample. Pressing the [START] key again starts
measurement of the next sample. Starting from the
1st sample, measurement can be made until the
[STOP] key is pressed or up to 150 samples.
Pressing the <1> key (Curve Confirmation) or <2> key
(Remeasure) allows the processing shown in Table
3-43.

3 - 109
3.3 Sample-by-sample Measurement (6-cell manual mode)

Photometry CELL 1

Sample name ID ABS CONC

1 0.15 15.04
Sample
concentration

Measured absorbance

START:Meas.SAMP.2; Select FUNC..：＿

1 Curve Confirmation □
□ 2 Remeasure

Fig. 3-71 Sample Measurement Screen

(3) When measurement is stopped by pressing the

[STOP]key or 150 samples have been measured, the
screen shown in Fig. 3-72 appears. For the
processing function assigned to each key, refer to
Table 3-45.

Photometry CELL 1
ID ABS CONC
11 0.001 0.000
12 0.102 10.10
13 0.205 20.05
14 0.172 17.04
15 0.051 5.10

Select function. : _
1 Meas. □
□ 2 Curve □ 3 Save □
4 Print

Example of display after measurement

of 15 samples

Fig. 3-72 Screen after Measurement of All Samples

3 - 110
 3.3.1

Table 3-45 Guidance after Measurement of All

Samples

Setting Item Description

[1] Meas. You can make setting related to
measurement. “New Meas.”, “Continue”
and “Remeasure” are selectable.
[1] New Meas. : Selected when newly
measuring samples
with the same
calibration curve.
[2] Continue : Allows you to start
measurement from the
number following the
previously measured
sample. When this
function is selected, the
guidance given below
appears. Enter the
number of samples to
be measured and press
the [ENTER] key.
Input value. ： ＿
No.of Sample [1-140]

Display With 10 Samples Measured

[3] Remeasure : Selected when

remeasuring a
measured sample.
When this function is
selected, the guidance
given below appears.
Select a sample to be
remeasured with the
[▲] or [▼] key, set the
sample to cell 1 and
press the [START] key
to carry out remeasure-
ment. To execute
auto zero, press the
[AUTOZERO] key.
Photometry CELL A
ID ABS CONC
6 0.301 30.00
7 0.251 25.11
8 0.419 40.90
9 0.110 11.00
10 0.234 23.50

Set sample to Cell1(Sample10).: _

START:Meas.; AUTOZERO:Autozero
◄►( 2/ 2)

3 - 111
3.3 Sample-by-sample Measurement (6-cell manual mode)

(cont’d)
Setting Item Description
[2] Curve You can generate a calibration curve with
the measured standards and confirm it.
When this function is selected, the
following screen appears.
Photometry/Curve CELL A
R = 0.9999, R2 = 0.9999
+
0.893 +
+
+
ABS +
+
+
0.001 +
+
0.000 CONC 90.000
Select function. : _
1 Scale □
□ 2 Factor □ 3 STD

For details of the function, refer to section

4.3.
[1] Scale : Allows you to change the
scale of calibration curve.
[2] Factor : Indicates the factor,
correlation coefficient and
determination coefficient of
calibration curve.
[3] STD : Indicates the result of
standard solution
measurement. Also allows
remeasurement of a
standard solution, deletion
of standard data
unnecessary for generation
of a calibration curve or
retrieval of deleted data.
[3] Save You can save the measured data.
When this function is selected, the
following screen appears.
Save Data CELL A
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
1 Save □
□ 2 Sort
◄ ► ( 1/ 1)

[1] Save : Allows you to save data under

a new file name.
[2] Sort : Allows you to rearrange the
currently saved data in the
order of file names or dates.

3 - 112
 3.3.1

(cont’d)
Setting Item Description
[4] Print You can select items to be printed.
When this function is selected, the screen
given below appears. For each item,
select whether to print out (ON or OFF).
Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Curve ON
STD ON
Sample ON

Select function. : _
1 ON □
□ 2 OFF

3 - 113
3.3 Sample-by-sample Measurement (6-cell manual mode)

3.3.2 Measuring Absorbance/Transmittance

This instrument is used for generating a calibration curve and determining the concentration
of an unknown sample, and for entering a calibration factor and determining the
concentration.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Setting of Measuring Conditions

*
3. Setting of Sample Conditions

* For steps 2 and 3, refer to section 3.2.2..

4. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <3> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The 6-cell condition (6 Cell) screen (Fig. 3-73)

appears.

3 - 114
 3.3.2

Photometry CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample SAMP.Autozero ON
3 6 Cell Autozero int. 5
4 Print No.of Sample 1
5 Save Method
6 Meas.Screen

Select function. : _
1 Auto □
□ 2 Manual

Fig. 3-73 6-cell Condition (6 Cell) Screen

(3) To set each item on the 6-cell condition (6 Cell)

screen, press the [ENTER] key. Or select each item
with the [▲] or [▼] key and make your setting
according to the guidance. For details of each
parameter, refer to Table 3-46.

After selecting “Manual” for 6 Cell Mode, return to the

Photometry screen by pressing the [RETURN] key or [◄]
key .

Table 3-46 “6 Cell” Setting Parameter

Setting Item Description

6 Cell Mode Specify a 6-cell operation mode for measurement.
[1] Auto
In this mode, a sample is set at each position
of the 6-cell holder beforehand and the holder
is automatically turned to automatically
perform from auto zero to standard/sample
measurement. It is recommended to use this
mode if you have 6 cells and there are many
samples to be measured. (For details of the
Auto mode, refer to section 3.2.2..)
[2] Manual
In this mode, standard/sample measurement
is carried out by using one position (cell 1 to 5
selectable) of the 6-cell holder. To eliminate
the influence of instrument drift (baseline
fluctuation) during measurement, auto zero
can easily be executed by setting a solution
for auto zero to cell A. In normal usage, set
a standard or sample solution to cell 1.

3 - 115
3.3 Sample-by-sample Measurement (6-cell manual mode)

5. Setting of Printing Conditions

*
6. Saving of Measuring Conditions

* For steps 5 and 6, refer to section 3.2.2..

Upon completion of setting, press the <6> key (Meas. Screen)
and then [ENTER] key or press the [MEAS. SCREEN] key to
determine the measuring conditions.

7. Measurement of Sample Solutions

(1) The auto zero execution screen (Fig. 3-74) appears.

Set an auto zero sample to cell A. Auto zero is
executed by pressing the [AUTOZERO] key.

Photometry 600.0 nm 0.000 ABS CELL A

[Info]
Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Fig. 3-74 Auto Zero Execution Screen

(2) The sample measurement screen (Fig. 3-75) appears.

Set the sample indicated in the guidance area in the
sample compartment. Pressing the [START] key
starts measurement at the current cell position.

NOTE: In 6-cell manual mode, the photometric value at

the cell position indicated on the screen is
measured. Before measurement, therefore,
check if the cell position where the sample has
been set coincides with the cell position
indicated on the screen.

3 - 116
 3.3.2

Photometry CELL 1
ID ABS
1

START:Meas. Sample1：＿

Fig. 3-75 Sample Measurement Screen

Auto zero can be executed by pressing the

[AUTOZERO] key when the guidance on sample
setting is indicated. For details, refer to Fig. 3-76.

<1> Check that a guide message “START: Meas.

SampleXX:_” is displayed. For carrying out auto zero,
auto zero sample should be set to cell A.
Photometry CELL 1

ID ABS CONC
1

START:Meas.Sample1:：＿

Sample Measurement Screen

<2> Set an auto zero sample, close the cover and press the
[AUTOZERO] key. The 6-cell holder will move to the
position of cell A and auto zero will be executed there
<3> After execution of auto zero, the 6-cell holder moves to
the position before the execution and the sample
measurement screen reappears.
GUIDE: For executing auto zero frequently, set an auto zero
sample to the position of cell A beforehand. It is
now possible to move to the position of cell A and
execute auto zero just by pressing the [AUTOZERO]
key.

Fig. 3-76 How to Execute Auto Zero

3 - 117
3.3 Sample-by-sample Measurement (6-cell manual mode)

(3) After measurement of a sample, its measured

absorbance is indicated at the corresponding sample
position on the screen. This screen appears after
measurement of each sample. Pressing the [START]
key again starts measurement of the next sample.
Starting from the 1st sample, measurement can be
made until the [STOP] key is pressed or up to 150
samples. Pressing the <1> key (Remeasure) allows
remeasurement of the last-measured sample.

Photometry CELL 1
Sample name ID ABS
1 0.15

Measured
absorbance
START:Meas.SAMP.2; Select FUNC..：＿
1 Remeasure
□

Fig. 3-77 Sample Measurement Screen

(4) When measurement is stopped by pressing the [STOP]

key or 150 samples have been measured, the screen
shown in Fig. 3-78 appears. For the processing
function assigned to each key, refer to Table 3-47.

Photometry CELL 1
ID ABS
11 0.001
12 0.102
13 0.205
14 0.172
15 0.051

Select function. : _
1 Meas. □
□ 2 Save □ 3 Print

Example of display after measurement

of 15 samples

Fig. 3-78 Screen after Measurement of All Samples

3 - 118
 3.3.2

Table 3-47 Guidance after Measurement of All

Samples

Setting Item Description

[1] Meas. You can make setting related to
measurement. “New Meas.”, “Continue”
and “Remeasure” are selectable.
[1] New Meas. : Selected when newly
measuring samples
with the same
calibration curve.
[2] Continue : Allows you to start
measurement from the
number following the
previously measured
sample. When this
function is selected, the
guidance given below
appears. Enter the
number of samples to
be measured and press
the [ENTER] key.
Input value. ： ＿
No.of Sample [1-140]

Display with 10 Samples Measured

[3] Remeasure : Selected when

remeasuring a
measured sample.
When this function is
selected, the guidance
given below appears.
Select a sample to be
remeasured with the
[▲] or [▼] key, set the
sample to cell 1 and
press the [START] key
to carry out remeasure-
ment. To execute
auto zero, press the
[AUTOZERO] key.
Photometry CELL A
ID ABS
6 0.301
7 0.251
8 0.419
9 0.110
10 0.234

Set sample to Cell1(Sample10).: _

START:Meas.; AUTOZERO:Autozero
◄►( 2/ 2)

3 - 119
3.3 Sample-by-sample Measurement (6-cell manual mode)

(cont’d)
Setting Item Description
[2] Save You can save the measured data.
When this function is selected, the
following screen appears.
Save Data CELL A
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
1 Save □
□ 2 Sort
◄ ► ( 1/ 1)

[1] Save : Allows you to save data under

a new file name.
[2] Sort : Allows you to rearrange the
currently saved data in the
order of file names or dates.
[3] Print You can select items to be printed.
When this function is selected, the screen
given below appears. For each item,
select whether to print out (ON or OFF).
Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Curve ON
Sample ON

Select function. : _
1 ON □
□ 2 OFF

3 - 120
 3.3.3

3.3.3 Measuring DNA Sample (measurement through ratio calculation)

This instrument is capable of measuring the absorbance (at

230 nm, 260 nm, 280 nm and 320 nm) of a DNA sample and calculating the absorbance ratio
(A260/A280 and A260/A230) for check of the DNA purity. The instrument is also used for
measuring the absorbance at 2 wavelengths and calculating the absorbance ratio or
absorbance difference between the values thus obtained.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Setting of Measuring Conditions

*
3. Setting of Sample Conditions

* For steps 2 and 3, refer to section 3.2.3..

4. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <3> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The 6-cell condition (6 Cell) screen (Fig. 3-79)

appears.

3 - 121
3.3 Sample-by-sample Measurement (6-cell manual mode)

Photometry CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample SAMP.Autozero ON
3 6 Cell Autozero int. 5
4 Print No.of Sample 1
5 Save Method
6 Meas.Screen

Select function. : _
1 Auto □
□ 2 Manual

Fig. 3-79 6-cell Condition (6 Cell) Screen

(3) To set the 6-cell conditions, press the [ENTER] key.

Or select each item with the [▲] or [▼] key and make
your setting according to the guidance. For details of
each parameter, refer to Table 3-48.

(4) After selecting “Manual” for 6 Cell Mode, return to the

Photometry screen by pressing the [RETURN] key or
[◄] key .

Table 3-48 “6 Cell” Setting Parameter

Setting Item Description

6 Cell Mode Specify a 6-cell operation mode for
measurement.
1) Auto
In this mode, a sample is set at each
position of the 6-cell holder beforehand
and the holder is automatically turned to
automatically perform from auto zero to
standard/sample measurement. It is
recommended to use this mode if you
have 6 cells and there are many samples
to be measured. (For details of the Auto
mode, refer to section 3.3.3..)
2) Manual
In this mode, standard/sample
measurement is carried out by using one
position (cell 1 to 5 selectable) of the 6-cell
holder. To eliminate the influence of
instrument drift (baseline fluctuation)
during measurement, auto zero can easily
be executed by setting a solution for auto
zero to cell A. In normal usage, set a
standard or sample solution to cell 1.

3 - 122
 3.3.3

5. Setting of Printing Conditions

*
6. Saving of Measuring Conditions

* For steps 5 and 6, refer to section 3.3.3..

Upon completion of setting, press the <6> key (Meas. Screen)
and then [ENTER] key or press the [MEAS. SCREEN] key to
determine the measuring conditions.

7. Measurement of Sample Solutions

(1) The auto zero execution screen (Fig. 3-80) appears.

Set an auto zero sample to cell A. Auto zero is
executed by pressing the [AUTOZERO] key.

Photometry 600.0 nm 0.000 ABS CELL A

[Info]
Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Fig. 3-80 Auto Zero Execution Screen

(2) The sample measurement screen (Fig. 3-81) appears.

Set the sample indicated in the guidance area in the
sample compartment. Pressing the [START] key
starts measurement at the current cell position.

NOTE: In 6-cell manual mode, the photometric value at

the cell position indicated on the screen is
measured. Before measurement, therefore,
check if the cell position where the sample has
been set coincides with the cell position
indicated on the screen.

3 - 123
3.3 Sample-by-sample Measurement (6-cell manual mode)

Photometry CELL 1
ID ABS
1 WL1 （ 260.0 nm）
WL2 （ 280.0 nm）

ABS DIFF=
ABS Ratio=

START:Meas. Sample1：＿

Fig. 3-81 Sample Measurement Screen

Auto zero can be executed by pressing the

[AUTOZERO] key when the guidance on sample
setting is indicated. For details, refer to Fig. 3-82.

<1> Check that a guide message “START: Meas.

SampleXX:_” is displayed. For carrying out auto zero,
auto zero sample should be set to cell A.
Photometry CELL 1
ID
10 WL1 （ 260.0nm） 0.423
WL2 （ 280.0nm） 0.268

ABS DIFF= 0.155

ABS Ratio= 1.582

START:Meas.Sample11:＿
1 Remeasure
□
◄ ► ( 10/ 11)

Sample Measurement Screen

<2> Set an auto zero sample, close the cover and press the
[AUTOZERO] key. The 6-cell holder will move to the
position of cell A and auto zero will be executed there
<3> After execution of auto zero, the 6-cell holder moves to
the position before the execution and the sample
measurement screen reappears.
HINT: For executing auto zero frequently, set an auto zero
sample to the position of cell A beforehand. It is now
possible to move to the position of cell A and execute
auto zero just by pressing the [AUTOZERO] key.

Fig. 3-82 How to Execute Auto Zero

3 - 124
 3.3.3

(3) After measurement of a sample, its absorbance is

indicated on the screen. This screen appears after
measurement of each sample. Pressing the [START]
key again starts measurement of the next sample.
Starting from the 1st sample, measurement can be
made until the [STOP] key is pressed or up to 150
samples. Pressing the <1> key (Remeasure) allows
remeasurement of the last-measured sample.

Sample No. Photometry CELL 1

ID ABS
3 WL1 （ 260.0 nm） 0.421
WL2 （ 280.0 nm） 0.269
Absorbance
ABS DIFF= 0.152
ABS Ratio= 1.565 Calculated absorbance difference
and absorbance ratio
START:Meas.SAMP.4; Select FUNC..：＿
1 Remeasure
□
◄►( 3/ 4)

Fig. 3-83 Sample Measurement Screen

(4) When measurement is stopped by pressing the [STOP]

key or 150 samples have been measured, the screen
shown in Fig. 3-84 appears. For the processing
function assigned to each key, refer to Table 3-49.

Photometry CELL 1
ID ABS
10 WL1 （ 260.0 nm） 0.423
WL2 （ 280.0 nm） 0.268

ABS DIFF= 0.155

ABS Ratio= 1.582

Select function. : _
1 Meas. □
□ 2 Save □ 3 Print
◄ ► ( 10/ 10)

Example of display after measurement

of 10 samples

Fig. 3-84 Screen after Measurement of All Samples

3 - 125
3.3 Sample-by-sample Measurement (6-cell manual mode)

Table 3-49 Guidance after Measurement of All

Samples

Setting Item Description

[1] Meas. You can make setting related to
measurement. “New Meas.”, “Continue” and
“Remeasure” are selectable.
[1] New Meas. : Selected when newly
measuring samples with
the same calibration curve.
[2] Continue : Allows you to start
measurement from the
number following the
previously measured
sample. When this
function is selected, the
guidance given below
appears. Enter the
number of samples to be
measured and press the
[ENTER] key.
Input value. ： ＿
No.of Sample [1-140]

Display with 10 Samples Measured

[3] Remeasure : Selected when

remeasuring a measured
sample. When this
function is selected, the
guidance given below
appears. Select a
sample to be remeasured
with the [▲] or [▼] key, set
the sample to the cell
position indicated on the
screen and press the
[START] key to carry out
remeasure-ment. To
execute auto zero, press
the [AUTOZERO] key.
Photometry CELL 1
ID ABS
10 WL1 （ 260.0 nm） 0.423
WL2 （ 280.0 nm） 0.268

ABS DIFF= 0.155

ABS Ratio= 1.582

Set sample and press the START key.:

◄ ► ( 10/ 10)

3 - 126
 3.3.3

[2] Save You can save the measured data. When this
function is selected, the following screen
appears.
Save Data CELL A
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
1 Save □
□ 2 Sort
◄ ► ( 1/ 1)

[1] Save : Allows you to save data under a

new file name.
[2] Sort : Allows you to rearrange the
currently saved data in the order
of file names or dates.
[3] Print You can select items to be printed.
When this function is selected, the screen
given below appears. For each item, select
whether to print out (ON or OFF).
Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Sample ON

Select function. : _
1 ON □
□ 2 OFF

3 - 127
3.3 Sample-by-sample Measurement (6-cell manual mode)

3.3.4 Measuring a Spectrum

This instrument is capable of measuring the transmission spectrum and absorption spectrum.
By setting the blank to cell A, one sample can be measured after automatic execution of
baseline correction.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Setting of Measuring Conditions

*
3. Setting of Sample Conditions

* For steps 2 and 3, refer to section 3.2.4..

4. Setting of 6-cell Conditions

NOTE: “6 Cell Mode” is not indicated if OFF is specified

for Mode on the 6-cell mode setting screen for
use of the single cell holder or rectangular long
path cell holder. (For details, refer to section
2.5.4..) When the auto sipper is connected,
“Sipper” is indicated in place of “6 Cell Mode”.

(1) To set the 6-cell conditions, press the <3> key (6 Cell)
and then [ENTER] key. Or select “6 Cell” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

3 - 128
 3.3.4

(2) The 6-cell condition (6 Cell) screen (Fig. 3-85)

appears. To set each item on the 6-cell condition (6
Cell) screen, press the [ENTER] key. Or use the [▲]
or [▼] key. Select “Manual” for 6 Cell Mode and
specify the number of samples to be measured for No.
of Sample. For details of each parameter, refer to
Table 3-50.

WL Scan CELL A
1 Meas.PARAM. 6 Cell Mode Auto
2 Sample No.of Sample 1
3 6 Cell
4 System
5 Peak PARAM.
6 Print
7 Save Method
8 Meas.Screen

Input character. :
Sample Name [Max: 8 characters]

Fig. 3-85 6-cell Condition (6 Cell) Screen

Table 3-50 “6 Cell” Setting Parameter

Setting Item Description
6 Cell Mode Select a 6-cell operation mode for
measurement from the following.
1) Auto
By setting a baseline sample and
unknown sample to the 6-cell holder, it
is possible to automatically turn the 6-
cell holder and automatically measure
one sample after baseline correction.
(For details of the Auto mode, refer to
section 3.2.4..)
2) Manual
Baseline correction and sample
measurement are performed manually.
It is required to manually move the cells
and execute baseline correction.

5. Setting of System Conditions

6. Setting of Peak Detection Conditions

*
7. Setting of Printing Conditions

8. Saving of Measuring Conditions

* For steps 5 to 8, refer to section 3.2.4..

3 - 129
3.3 Sample-by-sample Measurement (6-cell manual mode)

9. Baseline Correction

(1) To determine the measuring conditions set in the

preceding steps, press the <8> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key.

(2) The guidance on baseline correction (Fig. 3-86)

appears. Set a sample for baseline correction to cell
A. Upon pressing the [START] key, baseline
correction is carried out. If there is a baseline saved
under the same measuring conditions, you can also
proceed to sample measurement without performing
baseline correction with the CLEAR key.

WL Scan CELL A
Baseline CORR.

[Info]
Set baseline sample to Cell A.

START:Baseline CORR.

Fig. 3-86 Guidance on Baseline Correction

10. Measurement of Sample Solutions

(1) After completion of baseline correction, the sample

measurement screen (Fig. 3-87) appears. Set a
sample at the current cell position and press the
[START] key. Measurement will then start.
For performing baseline correction, press the
[RETURN] key.

3 - 130
 3.3.4

WL Scan CELL A
0.800

ABS

0.000
400.0 525.0 650.0 nm
START:Meas.Sample.
RETURN:Execute baseline CORR.

Fig. 3-87 Sample Measurement Screen

(2) After completion of measurement, the screen shown in

Fig. 3-88 appears. Pressing the <1> to <5> keys
allows the processings given in Table 3-51.
For proceeding to the next sample measurement,
press the [START] key.

WL Scan CELL A

0.800

ABS

0.000
400.0 525.0 650.0 nm
START:Meas.; Select function.：_
1 Data □
□ 2 Scale □3 Save □4 Print □
5 Baseline

Fig. 3-88 Screen after Sample Measurement

3 - 131
3.3 Sample-by-sample Measurement (6-cell manual mode)

Table 3-51 Guidance after Measurement in WL Scan

Mode

Setting Item Description

[1] Data You can perform each data processing.
When this function is selected, the following
guidance appears.
Select function. : _
[1]Trace [2]Peak [3]Smoothing [4]List

For details of the function, refer to section

4.3.
[1] Trace : Allows you to read out all
the measured data on the
spectrum.
[2] Peak : Displays the detected
peak.
[3] Smoothing : Allows you to perform
spectrum smoothing.
[4] List : Allows you to display the
measured data at the
specified intervals.
[2] Scale You can change the scale on the X axis and
Y axis of spectrum.
For detail of this function, refer to section 4.3.
[3] Save You can save the measured spectrum.
When this function is selected, the following
screen appears.
Save Data CELL A
1 NO3-(Hitachi Taro) WLS 2010/10/10
2 Cr6+(Hitachi Hanako) WLS 2010/03/30

Select function. : _
1 Save □
□ 2 Sort
◄ ► ( 1/ 1)

[1[ Save : Allows you to save data under

a new file name.
[2] Sort : Allows you to rearrange the
currently saved data in the
order of file names or dates.

3 - 132
 3.3.4

(cont’d)
Setting Item Description
[4] Print You can select items to be printed.
When this function is selected, the screen
given below appears. For each item, select
whether to print out (ON or OFF).
Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Graph ON
Peak Table ON
Data List ON

Select function. : _
1 ON □
□ 2 OFF
◄ ► ( 1/ 1)

[5] Baseline You can execute baseline correction.

When this function is selected, the following
guidance appears.
Select FUNC. and execute at Cell A.:＿
[1]Yes [2]No

[1] Yes : Selected to execute baseline

correction. Set a sample for
baseline correction to cell A
and press the <1> key.
[2] No : Selected to return to the
previous screen without
executing baseline correction.

(3) Before proceeding to the next measurement, the

system displays the data deletion confirming screen
(Fig. 3-89). When data has already been saved or
printed, select the <1> key (Yes). To save or print
data, select the <2> key (No) and perform the save or
print operation.

Data will be deleted. Continue? :＿

[1] Yes [2]No

Fig. 3-89 Data Deletion Confirming Screen

(4) Pressing the [START] key starts spectrum

measurement for the next sample.
After measurement, you can execute the same
operation as in (1).

3 - 133
3.3 Sample-by-sample Measurement (6-cell manual mode)

3.3.5 Measuring Change with Time

This instrument is used for measuring the sample absorbance/transmittance change with
time. You can evaluate the sample deterioration or enzyme activity based on the
absorbance change thus obtained.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Setting of Measuring Conditions

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 3-90) will appear. To set each condition for time
scan, press the <3> key (Time Scan) and then
[ENTER] key. Or select “Time Scan” with the [▲] or
[▼] key and press the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 3-90 Main Menu Screen

(2) The Time Scan screen (Fig. 3-91) appears. To set

the measuring conditions, press the <1> key (Meas.
PARAM.) and then [ENTER] key. Or select “Meas.
PARAM.” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

3 - 134
 3.3.5

Time Scan CELL A

1 Meas.PARAM. Data Mode ABS
2 Sample WL(nm) 500.0
3 System Scan Time (s) 60
4 Print Upper Scale 1.000
5 Save Method Lower Scale 0.000
6 Meas.Screen Delay(s) 0

Select function. : _

Fig. 3-91 Time Scan Screen

(3) The measuring condition (Meas. PARAM.) screen (Fig.

3-92) appears. With the [▲] or [▼] key, set the data
mode (Data Mode), wavelength (WL), scan time (Scan
Time), Y-axis upper limit (Upper Scale), Y-axis lower
limit (Lower Scale) and initial delay (Delay).
For details of each parameter, refer to Table 3-52.

Time Scan CELL A

1 Meas.PARAM. Data Mode ABS
2 Sample WL(nm) 500.0
3 System Scan Time(s) 60
4 Print Upper Scale 1.000
5 Save Method Lower Scale 0.000
6 Meas.Screen Delay(s) 0

Select function. : _
1 ABS □
□ 2 %T

Fig. 3-92 Measuring Condition (Meas. PARAM.)

Screen

Table 3-52 “Meas. PARAM.” Setting Parameters

Setting Item Description

Data Mode Select a data mode for the Y axis.
[1] ABS : Used for measuring the
absorbance.
[2] %T : Used for measuring the
transmittance.
WL (nm) Enter a wavelength to be used for
measurement.
This parameter is specifiable in a range of
190.0 to 1100.0 nm in steps of 0.1 nm.

3 - 135
3.3 Sample-by-sample Measurement (6-cell manual mode)

(cont’d)
Setting Item Description
Scan Time (s) Specify a scan time.
This parameter is specifiable in a range of
60 to 99999 sec in steps of 1 sec.
The data interval differs depending on the
scan time. It is set as shown in the
following table.
Scan Time and Data Interval

Data Interval
Scan time (sec)
(sec)
60 ≤ Scan time < 100 1
100 ≤ Scan time < 200 1
200 ≤ Scan time < 350 1
350 ≤ Scan time < 1000 2
1000 ≤ Scan time < 2000 5
2000 ≤ Scan time < 5000 5
5000 ≤ Scan time < 10000 10
10000 ≤ Scan time < 99999 100

Upper Scale Enter a Y-axis upper limit for the spectrum

displayed.
This parameter is specifiable in the
following range.
ABS : -999.9 to 999.9
%T : -9.999 to 9.999
Lower Scale Enter a Y-axis lower limit for the spectrum
displayed.
This parameter is specifiable in the
following range.
ABS : -999.9 to 999.9
%T : -9.999 to 9.999
Delay (s) After the [START] key is pressed, the
system waits for the time period set here
and then starts measurement.
This parameter is settable in a range of 0
to 9999 sec in steps of 1 sec.
Set this parameter when you want to start
measurement after a specified time period,
including when measuring a sample after
its temperature reaches the room
temperature and when starting
measurement after completion of reaction.
Enter “0” if the delay time need not be set.

(4) After completion of setting, return the Time Scan

screen by pressing the [RETURN] key or [◄] key.

3 - 136
 3.3.5

3. Setting of Sample Conditions

(1) To set the sample conditions, press the <2> key

(Sample) and then [ENTER] key. Or select “Sample”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The sample condition (Sample) screen (Fig. 3-93)

appears.

Time Scan CELL A

1 Meas.PARAM. Sample Name
2 Sample
3 System
4 Print
5 Save Method
6 Meas.Screen

Input character. :
Sample Name [Max: 8 characters]
ABC

Fig. 3-93 Sample Condition (Sample) Screen

(3) To enter/select the sample conditions, press the

[ENTER] key. Or select the item with the [▲] or [▼]
key and make your setting. For details of the
parameter, refer to Table 3-53.

(4) After entering a sample name, return to the Time Scan

screen by pressing the [ENTER] key or [◄] key.

Table 3-53 “Sample” Setting Parameter

Setting Item Description

Sample Name Up to 8 half-size alphanumeric characters
can be entered for the sample name.
The sample name entered here is printed in
the sample name field on the report.
For the character input procedure, refer to
section 2.4.3.

3 - 137
3.3 Sample-by-sample Measurement (6-cell manual mode)

4. Setting of System Conditions

(1) To set the system conditions, press the <3> key

(System) and then [ENTER] key. Or select “System”
with the [▲] or [▼] key and press the [ENTER] key or
[►] key.

(2) The system condition (System) screen (Fig. 3-94)

appears.

Time Scan CELL A

1 Meas.PARAM. Response Medium
2 Sample
3 System
4 Print
5 Save Method
6 Meas.Screen

Select function. : _
1 Fast □
□ 2 Medium □ 3 Slow

Fig. 3-94 System Condition (System) Screen

(3) Set up a response. After completion of setting, return

to the Time Scan screen by pressing the [RETURN]
key or [◄] key.

Setting Item Description

Response Select from the following 3 response speeds.
[1] Fast : Used for high-resolution
measurement with respect
to the wavelength.
Note that the noise level is
high as compared with
Medium and Slow.
[2] Medium : Used for ordinary
measurement.
[3] Slow : Used to reduce a variation
in photometric value.
This response speed is
unsuitable for sample
measurement reflecting an
abrupt change with time as
compared with Fast and
Medium.

3 - 138
 3.3.5

5. Setting of Printing Conditions

GUIDE: Specify print items if determined beforehand.

You can also specify print items after completion
of measurement. If you do not want to set the
printing conditions here, proceed to step 6.

(1) To set the printing conditions, press the <4> key (Print)
and then [ENTER] key. Or select “Print” with the [▲]
or [▼] key and press the [ENTER] key or [►] key.

(2) The printing condition (Print) screen (Fig. 3-95)

appears. To select the printing conditions, press the
[ENTER] key. Or select each item with the [▲] or [▼]
key and make your setting according to the guidance.
For details of each parameter, refer to Table 3-54.

Time Scan CELL A

1 Meas.PARAM. Auto Print OFF
2 Sample Print Date ON
3 System Run Date ON
4 Print Method ON
5 Save Method Graph ON
6 Meas.Screen Data List OFF

Select function. : _

Fig. 3-95 Printing Condition (Print) Screen

(3) After making your selection (ON or OFF) for Auto Print,
Print Date, Run Date, Method, Graph and Data List,
return to the Time Scan screen by pressing the
[RETURN] key or [◄] key.

3 - 139
3.3 Sample-by-sample Measurement (6-cell manual mode)

Table 3-54 “Print” Setting Parameters

Setting Location in
Description
Item Fig. 3-98
Auto Print [1] ON : Automatic printing is ⎯
carried out after
measurement.
[2] OFF : Automatic printing is
not carried out after
measurement.
Print Date [1] ON : Printing date is <1>
printed.
[2] OFF : Printing date is not
printed.
Run Date [1] ON : Analysis date is <2>
printed.
[2] OFF : Analysis date is not
printed.
Method [1] ON : Measuring conditions <3>
are printed.
[2] OFF : Measuring conditions
are not printed.
Graph [1] ON : Graph of change with <4>
time is printed.
[2] OFF : Graph of change with
time is not printed.
Data List [1] OFF : Numerical data of <5>
change with time
is not printed.
[2] All Data : Numerical data of
change with time
is all printed.
[3] Interval : Numerical data of
change with time
is printed at the
specified intervals.
Int. Specify a printing interval for the ⎯
data list. This parameter is
indicated when “Interval” is
selected for Data List. It is
specifiable in a range of 1 to 100.

3 - 140
 3.3.5

Report ： 2010/04/10 15:12 <1>

0.1

<4>

-0.1

0 15.0 30.0 45.0 60 s

Sample Name：
File Name：
Run Date： 2010/04/10 13:34
<2>
Operator：

Spectrophotometer
Model： U-5100 Spectrophotometer
Serial No.： 1234-567
ROM Ver.： 3J25300
Option：

Instrument
Meas.Mode： Time Scan Response： Medium
Data Mode： %T Bandpass(nm)： 5.0
WL (nm)： 220.0 6 Cell Mode： Manual <3>
Scan Time (s)： 60
Delay (s)： 0

Data List(Int.(s): 1)
No. TIME(s) %T No. TIME(s) %T No. TIME(s) %T <5>
1 0 0.0 3 2 0.0 5 4 0.0
2 1 0.0 4 3 0.0 6 5 0.0

Fig. 3-96 Example of Printout in Time Scan Mode

Setting of the measuring conditions has now been

completed. For saving the set conditions, proceed to step
6. If they need not be saved, proceed to step 7.

3 - 141
3.3 Sample-by-sample Measurement (6-cell manual mode)

6. Saving of Measuring Conditions

GUIDE: When the set measuring conditions need not be

saved, proceed to step 7.

(1) To save the set measuring conditions, press the <5>

key (Save Method) and then [ENTER] key. Or select
“Save Method” with the [▲] or [▼] key and press the
[ENTER] key or [►] key.

(2) The measuring condition saving (Save Method) screen

(with saved data) (Fig. 3-97) appears.

Save Method CELL A

1 NO3-(Hitachi Taro) TMS 2010/10/10
2 Cr6+(Hitachi Hanako) TMS 2010/03/30

File name Saving

date
Measurement
mode
File Name：_
File Name [Max: 20 characters]
ABC ◄ ► ( 1/ 1)

Fig. 3-97 Measuring Condition Saving (Save Method)

Screen (with saved data)

(3) When other measuring conditions are saved in

advance, a list of saved measuring conditions appears
on the main menu. At this time, the file name,
measurement mode (PHT: Photometry, WLS:
Wavelength Scan, TMS: Time Scan) and saving date
are indicated.

(4) For the measuring conditions to be saved, enter a file

name within 20 characters and press the [ENTER] key.

3 - 142
 3.3.5

NOTE: • If a file of the same name as entered already

exists in the same measurement mode, the
following guidance appears.
Already exists. Overwrite it? :＿
[1]Yes [2]No

For overwriting, press the <1> key (Yes).

To avoid overwriting, press the <2> key (No)
and rename the file to be saved.
• A total of 50 method files can be saved for
all measurement modes.
• If 50 files are already registered, the
following message appears.
No.of files is full.

In this case, delete files referring to section

4.2.2 and retry saving.
• The method for which auto start (for details,
refer to section 4.5.) is set is preceded by
“*”.

(5) Return to the Time Scan screen by pressing the

[RETURN] key.

7. Measurement of Sample Solutions

(1) To determine the measuring conditions set in the

preceding steps, press the <6> key (Meas. Screen)
and then [ENTER] key. Or press the [MEAS.
SCREEN] key.

(2) The auto zero execution screen (Fig. 3-98) appears.

Set an auto zero sample to cell A. Auto zero is
executed by pressing the [AUTOZERO] key.
To avoid executing auto zero, press the [CLEAR] key.

3 - 143
3.3 Sample-by-sample Measurement (6-cell manual mode)

Photometry 600.0 nm 0.000 ABS CELL A

[Info]
Set autozero sample to Cell A.

AUTOZERO:Execute Autozero; CLEAR:Cancel

Fig. 3-98 Auto Zero Execution Screen

(3) After execution of auto zero, the sample measurement

screen (Fig. 3-99) appears. Set a sample at the
present cell position and press the [START] key to start
measurement. For carrying out auto zero, press the
[RETURN] key.

NOTE: In Time Scan mode, the photometric value at

the cell position indicated on the screen is
measured. Before measurement, therefore,
check if the cell position where the sample has
been set coincides with the cell position
indicated on the screen.

WL Scan CELL A
0.800

ABS

0.000
0 60.0 120 s
START:Meas.: _

Fig. 3-99 Sample Measurement Screen

By pressing the [AUTOZERO] key when a guide

message “START: Start Meas.:_” is displayed, auto
zero can be executed at the position of cell A.
For details, refer to Fig. 3-100.

3 - 144
 3.3.5

<1> Check that a guide message “START: Start Meas.:_” is

displayed. For carrying out auto zero, auto zero sample
should be set to cell A.
START:Meas.: _

Sample Measurement Screen

<2> Set an auto zero sample, close the cover and press the
[AUTOZERO] key. The 6-cell holder will move to the
position of cell A and auto zero will be executed there
<3> After execution of auto zero, the 6-cell holder moves to
the position before the execution and the sample
measurement screen reappears.
GUIDE: For executing auto zero frequently, set an auto zero
sample to the position of cell A beforehand. It is now
possible to move to the position of cell A and execute
auto zero just by pressing the [AUTOZERO] key.

Fig. 3-100 How to Execute Auto Zero

(5) After completion of measurement, the sample

measurement result screen (Fig. 3-101) appears.
Pressing the <1> to <4> keys allows the processings
given in Table 3-55. For proceeding to the next
sample measurement, press the [START] key.

Time Scan CELL 1

0.800

ABS

0.000
0 60.0 120 s
START:Meas.; Select function.：_
1 Data □
□ 2 Scale □3 Save □4 Print

Fig. 3-101 Sample Measurement Result Wcreen

3 - 145
3.3 Sample-by-sample Measurement (6-cell manual mode)

Table 3-55 Guidance after Sample Measurement

Item Description
[1] Data You can perform each data processing.
When this function is selected, the following
guidance appears.
Select function. : _
[1]Trace [2]Smoothing [3]List

For details of the function, refer to section 4.3.

[1] Trace : Allows you to read out all the
measured data on the
spectrum.
[2] Smoothing : Allows you to perform
smoothing of the measured
data.
[3] List : Allows you to display the
measured data at the
specified intervals.
[2] Scale You can change the scale on the X axis and Y
axis of measured data.
For detail of this function, refer to section 4.3.
[3] Save You can save the measured spectrum.
When this function is selected, the following
screen appears.
Save Data CELL A
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Select function. : _
1 Save □
□ 2 Sort
◄ ► ( 1/ 1)

[1] Save : Allows you to save data under a

new file name.
[2] Sort : Allows you to rearrange the
currently saved data in the order of
file names or dates.
[4] Print You can select items to be printed. When this
function is selected, the screen given below
appears. For Print Date, Run Date, Method
and Graph, select “ON” or “OFF”. For Data
List, select from OFF, All Data and Interval.
When “Interval” is selected, “Int.” is indicated.
Photometry CELL A
Print
Print Date ON
Run Date ON
Method ON
Graph ON
Data List Int.
Int.(s) 1

Select function. : _
1 ON □
□ 2 OFF

3 - 146
 3.4

3.4 Measurement with Enlarged Display Screen (monitor)

This instrument displays a photometric value in enlarged size on the monitor. This function
is effective in carrying out measurement while monitoring the photometric value at a
wavelength. Described below is the operating procedure for measurement with this function.

GUIDE: When auto start is specified, the system automatically sets up the conditions and
advances to the measurement screen phase of sequence after the power switch is
turned on. For how to specify auto start, refer to section 4.5..

1. Startup of This Product

Start this product. (For the starting procedure, refer to

section 2.3.1..)

2. Display of Main Menu

Press the [MAIN MENU] key. The Main Menu screen (Fig.
3-102) will then appear. To present the enlarged display
screen, press the <8> key (Monitor) and then [ENTER] key.
Or select “Monitor” with the [▲] or [▼]key and press the
[ENTER] key.

GUIDE:The enlarged display screen can also be presented

by pressing the [MEAS. SHIFT] key with the
[SHIFT] key held down.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 3-102 Main Menu Screen

3 - 147
3.4 Measurement with Enlarged Display Screen (monitor)

3. Presentation of Enlarged Display Screen (monitor)

The enlarged display screen (Fig. 3-103) appears. At this

time, the lamp turns on automatically and measurement
starts. In the upper part of the monitor, the current
wavelength setting is displayed, and a photometric value in
the lower part. This photometric value is updated at
intervals of 2.5 sec. To display the absorbance, press the
<1> key (ABS) and the <2> key (%T) to display the
transmittance, then press the [ENTER] key.
When changing the wavelength to be used for
measurement, press the [GO TO WL] key, enter a new
wavelength and press the [ENTER] key.

Data Display CELL 1

WL:

1100.0 nm
Data:

0.001 ABS

Select function. : _
1)ABS 2)%T 3)Lamp ON

Fig. 3-103 Enlarged Display Screen

The enlarged display screen is provided with the automatic

lamp off function, which serves to prevent the lamp staying
on unnecessarily for a long time. After the lapse of
automatic lamp off time, the lamp is automatically turned off
and “-.---” is indicated for the photometric value as shown in
Fig. 3-104. To display the photometric value again, press
the <3> key (Lamp ON) and then [ENTER] key.
For the auto lamp off time (Auto Lamp OFF), “5 min” is set
as default. For changing this setting, refer to section 2.5.3.

3 - 148
 3.4

Data Display CELL 1

WL:

1100.0 nm
Data:

-.--- ABS

Select function. : _
1)ABS 2)%T 3)Lamp ON

Fig. 3-104 Enlarged Display Screen with Lamp Off

4. Measurement of Sample

(1) Confirm the cell position display screen. When the 6-

cell holder is connected, move to the aimed-at cell
position by pressing the [◄] or [►] key with the
[SHIFT] key held down. Move to cell 1 unless
otherwise specified.

(2) Press the [Autozero] key. Execute auto zero

according to the guidance. When OFF is specified for
6 Cell Mode (when the single cell holder or rectangular
long path cell holder is mounted), set an auto zero
sample beforehand and press the [Autozero] key.

(3) When the 6-cell holder is connected, set a sample to

the cell position indicated on the screen. When the
single cell holder or rectangular long path cell holder is
in use, set a sample to the cell holder.

(4) Read out and record the photometric value displayed

on the screen.

NOTE: The photometric value displayed is updated at

intervals of 2.5 sec. Read out the updated
photometric value after the sample
compartment becomes ready for measurement
(when a sample has been set for example).

3 - 149
3.4 Measurement with Enlarged Display Screen (monitor)

(5) When the printer is connected, print the wavelength

and photometric value by pressing the [PRINT] key.

(Example of Printout)

500.0 nm 0.203 Abs

(6) After completion of measurement, return to the

previous screen by pressing the [RETURN] key.

3 - 150
 4.1.1

4. FOR MORE EFFICIENT OPERATION

Explained here is usage of the advanced-level functions for more

efficient operations of this product. Read this section so that
you can get a smoother control of the product.

4.1 Loading or Deleting the Saved Data

Described below are operation methods such as loading and

deletion of the saved data.

4.1.1 Loading the Saved Data

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Data Loading Procedure

(1) Press the [MAIN MENU] key. The Main Menu screen
(shown in Fig. 4-1) appears. Press the <5> key (for
Data Menu) and then the [ENTER] key or press the
[▲]/[▼] key to select Data Menu and then the [ENTER]
key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 4-1 Main Menu Screen

4-1
4.1 Loading or Deleting the Saved Data

(2) Data Menu screen (Fig. 4-2) appears. For loading the
saved data, press the <1> key (for Load/Delete Data)
and then [ENTER] key or press the [▲]/[▼] key to
select Load/Delete Data and then the [ENTER] key.

Data Menu CELL A

1 Load/Delete Data
2 Print Data List

Select function. :

Fig. 4-2 Data Menu Screen

(3) The Load/Delete Data screen (Fig. 4-3) appears.

On this screen, the saved data files are listed.
On each row, the file name, type and date of file saving
are shown from the left. “Type” denotes the type of
Method in an abbreviation listed in Table 4-1. Select
the data to be loaded by the [▲]/[▼] key and press the
<1> key (for Load). For other keys, refer to Table 4-2.

Load/Delete Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30
3 Stability TMS 2010/05/05
4 Vitamin A WLS 2010/02/24

File name Date of file

saving
Type
Select function. :_
[1]Load [2]Delete [3]Check [4]Type [5]Sort
◄ ► ( 1/ 1)

Fig. 4-3 Load/Delete Data Screen

Table 4-1 Abbreviation of Method Type

Type Abbreviation
Photometry PHT
Wavelength scan WLS
Time scan TMS

4-2
 4.1.1

Table 4-2 Guidance for Load/Delete Data Screen

Function Key Explanation

[1] Load Loads the file which is now selected.
[2] Delete Deletes the file which is now selected.
[3] Check Enables you to check the outline of the
Method in the presently selected file.
[4] Type Enables you to select the type of the file
to be displayed on the screen.
Used for displaying in only one type such
as photometry, wavelength scan or time
scan. In default, all types are set for
display.
[5] Sort Enables you to sort the files to be
displayed on the screen.
Sorting is allowed in the alphabetical
order of file names or in the chronological
order of saved files. The alphabetical
order of file names is selected by default.

(4) The data is now loaded. At this time, U-5100 is set to

same Method as read Data. (However, exclude it when
Data of Auto Sipper [another sales] was opened while
using 6- Cell holder and when 6- Cell holder is opened
while using Auto Sipper. )According to the guidance,
data processing, re-measurement, etc. can be carried
out.
Note ：Please keep the same and measure Method of 6- Cell
Mode or Auto sipper (another sales) when measuring it with Curve
and Method of read Data.

When 6- Cell Mode of read Data and device or Auto sipper method
is different, the following guidances are displayed.

Data using 6 Cell will be loaded.: _

□1 Yes □
2 No

When you read data that uses 6-Cell holder

Data using sipper will be loaded.: _
□1 Yes □
2 No

When you read Data that uses Auto sipper (another sales)

When Data is read, [1]Yes is selected. When Data is not read,

[2]No is selected. Moreover, please go in the remeasuring after
reviewing Method without fail when such a guidance is displayed.

4-3
4.1 Loading or Deleting the Saved Data

4.1.2 Deleting the Saved Data

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Data Deleting Procedure

(1) Press the [MAIN MENU] key. The Main Menu screen
(shown in Fig. 4-4) appears. Press the <5> key (for
Data Menu) and then the [ENTER] key or press the
[▲]/[▼] key to select Data Menu and then the [ENTER]
key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-4 Main Menu Screen

(2) Data Menu screen (Fig. 4-5) appears. For displaying

the saved data, press the <1> key (for Load/Delete
Data) and then [ENTER] key or press the [▲]/[▼] key
to select Load/Delete Data and then the [ENTER] key.

Data Menu CELL A

1 Load/Delete Data
2 Print Data List

Select function. :_

Fig. 4-5 Data Menu Screen

4-4
 4.1.2

(3) The Load/Delete Data screen (Fig. 4-6) appears.

On this screen, the saved data files are listed.
On each row, the file name, type and date of file saving
are shown from the left. “Type” denotes the type of
Method in an abbreviation listed in Table 4-3. Select
the data to be deleted by the [▲]/[▼] key and press
the <2> key (for Delete). The guidance for
confirmation of your deletion will be displayed.
When there is no problem, execute the deletion.
For other keys, refer to Table 4-4.

NOTE: Data, once deleted, cannot be recovered.

Careful confirmation is required before data
deletion.

Load/Delete Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30
3 Stability TMS 2010/05/05
4 Vitamin A WLS 2010/02/24

File name Date of file

saving
Type
Select function. :_
[1]Load [2]Delete [3]Check [4]Type [5]Sort
◄ ► ( 1/ 1)

Fig. 4-6 Load/Delete Data Screen

Table 4-3 Abbreviation of Method Type

Type Abbreviation
Photometry PHT
Wavelength scan WLS
Time scan TMS

4-5
4.1 Loading or Deleting the Saved Data

Table 4-4 Guidance for Load/Delete Data Screen

Function Key Explanation

[1] Load Loads the file which is now selected.
[2] Delete Deletes the file which is now selected.
[3] Check Enables you to check the outline of the
Method in the presently selected file.
[4] Type Enables you to select the type of the file
to be displayed on the screen.
Used for displaying in only one type such
as photometry, wavelength scan or time
scan. In default, all types are set for
display.
[5] Sort Enables you to sort the files to be
displayed on the screen.
Sorting is allowed in the alphabetical
order of file names or in the chronological
order of saved files. The alphabetical
order of file names is selected by default.

4-6
 4.1.3

4.1.3 Printing the List of Saved Data

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Display of Print List Screen

(1) Press the [MAIN MENU] key. The Main Menu screen
(shown in Fig. 4-7) appears. Press the <5> key (for
Data Menu) and then the [ENTER] key or press the
[▲]/[▼] key to select Data Menu and then the [ENTER]
key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-7 Main Menu Screen

(2) Data Menu screen (Fig. 4-8) appears. For printing the
data list, press the <2> key (for Print List) and then
[ENTER] key or press the [▲]/[▼] key to select Print
List and then the [ENTER] key.

Data Menu CELL A

1 Load/Delete Data
2 Print Data List

Select function. :_

Fig. 4-8 Data Menu Screen

4-7
4.1 Loading or Deleting the Saved Data

(3) The Print List screen (Fig. 4-9) appears. On this

screen, the saved data files are listed. On each row,
the file name, type and date of file saving are shown
from the left. “Type” denotes the type of Method in an
abbreviation listed in Table 4-5.

Load/Delete Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30
3 Stability TMS 2010/05/05
4 Vitamin A WLS 2010/02/24

File name Date of file

saving
Type
Select function. :_
[1]Load [2]Delete [3]Check [4]Type [5]Sort
◄ ► ( 1/ 1)

Fig. 4-9 Print List Screen

Table 4-5 Abbreviation of Method Type

Type Abbreviation
Photometry PHT
Wavelength scan WLS
Time scan TMS

3. Printing of Data List

For printing in only one type of Method, select the <2> key
(for Type) and then PHT, WLS or TMS. For changing the
display order of file names, select the <3> key (for Sort) and
then “File Name” or “Data and Time.” The set Method will
be reflected on display. When you press the [PRINT] key,
the files on display are printed in the order of display. For
details of other key operations, refer to the guidance for
Print List (Table 4-6). For an example of printout, see Fig.
4-10.

4-8
 4.1.3

Table 4-6 Guidance for Print List

Function Key Explanation

[1] Confirmation Enables you to confirm the outline of the Method in
the presently selected file.
[2] Type Enables you to select the type of the file to be
displayed on the screen (from among All,
Photometry, WL Scan and Time Scan).
In default, all types are set for display.
Select function.:_
[1] All, [2] Photometry, [3] WL Scan, [4] Time Scan

[3] Sort Enables you to sort the files to be displayed on the

screen.
Sorting is allowed in the alphabetical order of file
names (File Name) or in the chronological order of
saved files (Date and Time). The alphabetical
order of file names is selected by default.
Select function.:_
[1] File Name, [2] Date and Time

Data List
No. File Name Type Date
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30
3 Stability TMS 2010/05/05
4 Vitamin A WLS 2010/02/24

Fig. 4-10 Example of Print List

4-9
4.1 Loading or Deleting the Saved Data

4.1.4 Photometry Based on Saved Calibration Curve Data

1. Startup of this product

Start up this product. (For startup method, see section

2.3.1..)

2. Calibration Curve Data Loading Procedure

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 4-11) appears.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-11 Main Menu Screen

(2) For loading the saved calibration curve data, press the
<5> key (for Data Menu) and then the [ENTER] key or
press the [▲]/[▼] key to select Data Menu and then
the [ENTER] key.

(3) Data Menu screen (Fig. 4-12) appears.

Data Menu CELL A

1 Load/Delete Data
2 Print Data List

Select function. :_

Fig. 4-12 Data Menu Screen

4 - 10
 4.1.4

(4) For loading the saved calibration curve, press the <1>
key (for Load/Delete Data) and then the [ENTER] key
or press the [▲]/[▼] key to select Load/Delete Data
and then the [ENTER] key. The Load/Delete Data
screen (Fig. 4-13) appears.

Load/Delete Data CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

Date of file
File name
saving
Measurement
mode
Select function. :_
[1]Load [2]Delete [3]Check [4]Type [5]Sort
◄ ► ( 1/ 1)

Fig. 4-13 Load/Delete Data Screen

(5) On this screen, the saved data files are listed.

The photometric data including the calibration curve is
indicated by PHT in the column of measurement mode.
For checking the parameters according to which the
saved data had been measured, press the <3> key (for
Check) and then the [ENTER] key. The outline of the
selected Method is now displayed. After check, you
can return to the previous display by pressing the
[RETURN] key. Select the data, in which the desired
calibration curve is saved, by the [▲]/[▼] key and press
the [ENTER] key. The saved data will be displayed.

NOTE ： Please use the data that 6- Cell Mode measured by

turning on without fail for the reading measurement when you
measure 6- Cell Mode in the monitor configuration while turned on.
Please use data to measure 6- Cell Mode by turning off without fail
for the reading measurement when you measure 6- Cell Mode in
the monitor configuration while turned off. Please use Data
measured with Auto Sipper without fail for the reading
measurement when measuring it by using Auto Sipper (another
sales).

3. Sample Measurement

(1) The contents of data are displayed. When the

contents are only standard data, the list shown in Fig.
4-14 appears, and that shown in Fig. 4-15 appears

4 - 11
4.1 Loading or Deleting the Saved Data

when the data of sample measurement is included.

If wavelength scan or time scan data is opened, the
calibration curve data cannot be found.
Therefore, you must return to Step 2 “Calibration Curve
Data Loading Procedure” and select the data again.

[When the open data is standard data alone]

Photometry CELL 1
ID ABS CONC
STD1 0.001 0.000
STD2 0.102 10.00
STD3 0.205 20.00

Select function. :_
1)Sample Meas. 2)Curve 3)Save

Example of screen showing measured data of

3 standard samples

Fig. 4-14 Standard Data-only Screen

Sample measurement is allowed by selecting the <1>

key (for Sample).

[When the open data includes sample data]

Photometry CELL 1
ID ABS CONC
11 0.001 0.000
12 0.102 10.10
13 0.205 20.05
14 0.172 17.04
15 0.051 5.10

Select function. :_
1)Meas. 2)Curve 3)Save 4)Print

Example of screen showing measured data of

15 unknown samples

Fig. 4-15 Sample Data-included Screen

When you select the <1> key (for Meas.) and then the
<1> key (for New Meas.), the data area of the open file
is cleared and sample measurement is allowed with
the same calibration curve as that of this file.

4 - 12
 4.1.4

(2) Set a sample for auto zero and execute auto zero.
Then, measure the sample. Thus, sample
measurement can be performed with the saved
calibration curve.

4 - 13
4.2 Loading and Deleting the Saved Method

4.2 Loading and Deleting the Saved Method

Introduced here is the photometric method by way of loading the

saved Method.

4.2.1 Measurement via Loading of Saved Method

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Method Loading Procedure

(1) After completion of initial setting, the Main Menu

screen (Fig. 4-16) appears.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-16 Main Menu Screen

(2) For loading a Method, press the <4> key (for Method
Menu) and then the [ENTER] key or press the [▲]/[▼]
key to select Method Menu and then the [ENTER] key.

(3) Method Menu screen (Fig. 4-17) appears.

Meas.PARAM. CELL A
1 Load/Delete Method
2 Auto Start
3 Print Method List

Select function. :_

Fig. 4-17 Method Menu Screen

4 - 14
 4.2.1

(4) For loading a Method, press the <1> key (for

Load/Delete Data) and then the [ENTER] key or press
the [▲]/[▼] key to select Load/Delete Data and then
the [ENTER] key.

Load/Delete Method CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

File name Date of file

saving
Type

Select function. :_
[1]Load [2]Delete [3]Check [4]Type [5]Sort
◄ ► ( 1/ 1)

Fig. 4-18 Load/Delete Data Screen

(6) On this screen, the saved Methods are listed.

File Name, Type, and Saving Date are shown from the
left. Type shows the kind of Method. PHT and WL Scan
display WLS and Time Scan in Photometry by the code
address of TMS. Select the Method to be loaded by the
[▲]/[▼] key and press the [ENTER] key. The Method
of the selected file is read.In case the measurement
parameters of the selected file name are unknown, the
outline of the selected Method can be displayed by
pressing the <3> key (for Check) and then the [ENTER]
key. After check, you can return to the previous
display by pressing the [RETURN] key.

Note ： The guidance of [Please review method before use.]is

displayed when Method of 6- Cell holder was opened on the
condition that Auto Sipper (another sales) is installed in the device
and when Method of Auto Sipper is opened on the condition that 6-
Cell holder is installed in the device. This is peculiar Meas.PARAM
in preserved Method to Auto Sipper or 6- Cell holder . It is however
because of no preservation.

Please review method before use.: _

□1 Yes □
2 No

When this is understood, and Method is read, [1]Yes is

4 - 15
4.2 Loading and Deleting the Saved Method

selected. When Method is not read, [2]No is selected.

Please refer to Table 4-7 for details.

Table 4-7 Set state of U-5100 when measurement

condition is opened
Opening Method U-5100 Condition
6 Cell Mode ON 6 Cell Mode OFF Auto Sipper
(Available
Accessory)
6 Cell Mode ON Method is 6-Cell Mode of Method in the part
reflected. System Setup is other than Auto
changed to turning Sipper are
ON, and Method is reflected. The
reflected. condition
6 Cell Mode OFF 6-Cell Mode of Method is concerning Auto
System Setup is reflected. Sipper is not set to
changed to turning preserved Method,
OFF and Method is and review Method,
reflected. please before it
measures it.
Auto Sipper Method in the part other than 6-Cell Method is
(Available Mode are reflected. The condition reflected.
Accessory) concerning 6-Cell Mode is not set to
preserved Method, and review Method,
please before it measures it.

4 - 16
 4.2.2

4.2.2 Deletion of Saved Method

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Call-up of Load/Delete Method Screen

(1) Press the [MAIN MENU] key. The Main Menu screen
(Fig. 4-19) appears.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-19 Main Menu Screen

(2) For displaying a list of Methods, press the <4> key (for
Method Menu) and then the [ENTER] key or press the
[▲]/[▼] key to select Method Menu and then the
[ENTER] key.

(3) Method Menu screen (Fig. 4-20) appears. Press the

<1> key (for Load/Delete Data) and then the [ENTER]
key or press the [▲]/[▼] key to select Load/Delete
Data and then the [ENTER] key.

Meas.PARAM. CELL A
1 Load/Delete Method
2 Auto Start
3 Print Method List

Select function. :_

Fig. 4-20 Method Menu Screen

4 - 17
4.2 Loading and Deleting the Saved Method

3. Deletion of Method

The saved Methods are now listed on the screen.

On each row, the file name, type and date of file saving are
shown from the left. “Type” denotes the type of Method in
an abbreviation listed in Table 4-8. Select the data to be
deleted by the [▲]/[▼] key and press the <2> key (for
Delete). The guidance for confirmation of your deletion will
be displayed. When there is no problem, execute the
deletion. For other keys, refer to Table 4-9.

NOTE: Data, once deleted, cannot be recovered.

Careful confirmation is required before data
deletion.

Load/Delete Method CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30

File name Date of file

saving
Measurement
mode
Select function. :_
[1]Load [2]Delete [3]Check [4]Type [5]Sort
◄ ► ( 1/ 1)

Fig. 4-21 Load/Delete Method Screen

Table 4-8 Abbreviation of Method Type

Type Abbreviation
Photometry PHT
Wavelength scan WLS
Time scan TMS

4 - 18
 4.2.2

Table 4-9 Guidance for Load/Delete Method Screen

Function Key Explanation

[1] Load Loads the file which is now selected.
[2] Delete Deletes the file which is now selected.
[3] Check Enables you to check the outline of the Method in
the presently selected file.
[4] Type Enables you to select the type of the file to be
displayed on the screen.
Used for displaying in only one type such as
photometry, wavelength scan or time scan.
In default, all types are set for display.
[5] Sort Enables you to sort the files to be displayed on the
screen.
Sorting is allowed in the alphabetical order of file
names or in the chronological order of saved files.
The alphabetical order of file names is selected by
default.

4 - 19
4.2 Loading and Deleting the Saved Method

4.2.3 Printing the List of Saved Methods

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Display of Method List Screen

(1) Press the [MAIN MENU] key. The Main Menu screen
(shown in Fig. 4-22) appears. Press the <4> key (for
Method Menu) and then the [ENTER] key or press the
[▲]/[▼] key to select Method Menu and then the
[ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-22 Main Menu Screen

(2) Method Menu screen (Fig. 4-23) appears.

For printing the method list, press the <3> key (for Print
List) and then [ENTER] key or press the [▲]/[▼] key to
select Print List and then the [ENTER] key.

Main Menu CELL A

1 Load/Delete Method
2 Auto Start
3 Print Method List

Select function. :_

Fig. 4-23 Method Menu Screen

4 - 20
 4.2.3

(3) The Print List screen (Fig. 4-24) appears. On this

screen, the saved data files are listed. On each row,
the file name, type and date of file saving are shown
from the left. “Type” denotes the type of Method in an
abbreviation listed in Table 4-10.

Print Method List CELL A

1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30
3 Stability TMS 2010/05/05
4 Vitamin A WLS 2010/02/24

Date of file
File name saving
Type
Select function. :_
[1]Confirmation [2]Type [3]Sort
◄ ► ( 1/ 1)

Fig. 4-24 Print List Screen

Table 4-10 Abbreviation of Method Type

Type Abbreviation
Photometry PHT
Wavelength scan WLS
Time scan TMS

3. Printing of Method List

For printing in only one type of Method, select the <2> key
(for Type) and then PHT, WLS or TMS. For changing the
display order of file names, select the <3> key (for Sort) and
then “File Name” or “Data and Time” for display order.
The set Method will be reflected on display. When you
press the [PRINT] key, the files on display are printed in the
order of display. For details of other key operations, refer
to Table 4-11. For an example of printout, see Fig. 4-25.

4 - 21
4.2 Loading and Deleting the Saved Method

Table 4-11 Guidance for Print List

Function Key Explanation

[1] Confirmation Enables you to confirm the outline of the Method in
the presently selected file.
[2] Type Enables you to select the type of the file to be
displayed on the screen (from among All,
Photometry, WL Scan and Time Scan).
In default, all types are set for display.
Select function.: _
[1] All, [2] Photometry, [3] WL Scan, [4] Time Scan

[3] Sort Enables you to sort the files to be displayed on the

screen.
Sorting is allowed in the alphabetical order of file
names (File Name) or in the chronological order of
saved files (Date and Time). The alphabetical
order of file names is selected by default.
Select function.:_
[1] File Name, [2] Date and Time

Method List
No. File Name Type Date
1 NO3-(Hitachi Taro) PHT 2010/10/10
2 Cr6+(Hitachi Hanako) PHT 2010/03/30
3 Stability TMS 2010/05/05
4 Vitamin A WLS 2010/02/24

Fig. 4-25 Example of Print List

4 - 22
 4.3.1

4.3 Data Processing

Explanation is given here on data processing of calibration curve,

spectrum and time-scan measurement.

4.3.1 Scale Change of Calibration Curve

1. Display of Calibration Curve Data

[Confirmation from saved data]

For check by opening the saved data, open the photometric

data including the saved calibration curve data according to
“4.1.1 Loading the Saved Data.”
Upon opening the calibration curve data, a guidance (Fig.
4-26) is presented. Select the <2> key (for Curve).

Select function. :_
[1]Meas. [2]Curve [3]Save [4]Print
◄ ► ( 2/ 2)

Fig. 4-26 Example of Guidance upon

Opening of Calibration Curve Data

[During measurement]

During measurement by selecting Photometry in the Main

Menu, Conc for Data Mode and 1st Order for Curve Type,
select Curve Confirmation or Curve indicated in the
guidance.

4 - 23
4.3 Data Processing

2. Scale Changing Procedure

(1) Calibration curve data is now opened. Select the <1>

key (for Scale).

Photometry/Curve CELL A
R = 0.9999 R2 = 0.9999
0.893 +
+
+
+
ABS +
+
+
0.001 + +
0.000 CONC 90.000

Select function. :_
[1]Scale [2]Factor [3]STD

Fig. 4-27 Calibration Curve Data

(2) Scale Change screen (Fig. 4-28) appears. On this

screen, a scale change, scale resetting and auto scale
operation are allowed. For each parameter (setting
item), refer to Table 4-12 “Scale Changing
Parameters.”

Photometry/Curve CELL A
Scale Change
X-Max 90.00
X-Min 0.001
Y-Max(ABS) 0.893
Y-Min (ABS) 0.001
Scale Reset
Auto Scale

Input value. ： ＿
X-Max[-9999 - 9999]

Fig. 4-28 Scale Change Screen

4 - 24
 4.3.1

Table 4-12 Scale Changing Parameters

Parameter Explanation
X-Max Sets the maximum value on the X axis
(representing the concentration of standard
solution) of calibration curve.
Settable within -9999 to -0.001, 0, 0.001 to
9999 (indication in max. 4 digits).
Change on the set axis is adopted by
pressing the [RETURN] key.
X-Min Sets the minimum value on the X axis
(representing the concentration of standard
solution) of calibration curve.
Settable within -9999 to -0.001, 0, 0.001 to
9999 (indication in max. 4 digits).
Change on the set axis is adopted by
pressing the [RETURN] key.
Y-Max (ABS) Sets the maximum value on the Y axis
(representing the absorbance) of calibration
curve.
Settable within -9.999 to 9.999.
Change on the set axis is adopted by
pressing the [Return] key.
Y-Min (ABS) Sets the minimum value on the Y axis
(representing the absorbance) of calibration
curve.
Settable within -9.999 to 9.999.
Change on the set axis is adopted by
pressing the [Return] key.
Scale Reset Sets the default value of scale.
For setting, press the <1> key (Execute).
Select function. :_
[1] Execute

Auto Scale Judges the maximum and minimum values of

the measured data on the Y axis, and
automatically optimizes the scale.
For setting, press the <1> key (Execute).
Select function. :_
[1] Execute

4 - 25
4.3 Data Processing

4.3.2 How to Check Calibration Curve Factor, Correlation Coefficient and

Determination Coefficient

1. Display of Calibration Curve Data

[Confirmation from saved data]

For check by opening the saved data, open the photometric

data including the saved calibration curve data according to
“4.1.1 Loading the Saved Data.”
Upon opening the calibration curve data, a guidance (an
example shown in Fig. 4-29) is presented. Select the <2>
key (for Curve).

Select function. :_
[1]Meas. [2]Curve [3]Save [4]Print
◄►( 2/ 2)

Fig. 4-29 Example of Guidance upon

Opening of Calibration Curve Data

[During measurement]

During measurement by selecting Photometry in the Main

Menu, Conc for Data Mode and 1st Order for Curve Type,
select Curve Confirmation or Curve indicated in the
guidance.

2. Curve Factor Checking Procedure

(1) The calibration curve data is now opened. Select the

<2> key (for Factor).

Photometry/Curve CELL A
R = 0.9999 R2 = 0.9999
+
0.893 +
+
+
ABS +
+
+
0.001 + +
0.000 CONC 90.000
Select function. :_
[1]Scale [2]Factor [3]STD

Fig. 4-30 Calibration Curve Data

4 - 26
 4.3.2

(2) Calibration Curve Factor Confirmation screen (Fig. 4-

31) appears. On this screen, calibration curve factor,
correlation coefficient and determination coefficient can
be set. For each parameter (setting item), refer to
Table 4-13.

Photometry/Curve CELL A
Curve Factor
ABS = f(CONC)
A0 = 0
A1 = 1.000
Correlation Coef.
R = 0.9999
Determination Coef.
R2 = 0.9999

Fig. 4-31 Calibration Curve Factor

Confirmation Screen

Table 4-13 Explanation of Calibration Curve Factor

Confirmation Screen

Parameter Explanation
Curve Factor Denotes the factor of calibration curve.
A0 and A1 stand for the values expressed by
the following equation.
[When ABS = f(CONC)]
Absorbance = A1 x concentration + A0
Concentration = (absorbance - A0)/A1
[When CONC = f(ABS)]
Concentration = A1 x absorbance + A0
Correlation Coef. Used for evaluating the linearity of calibration
curve. When this value is closer to 1.0000,
the linearity can be evaluated to be higher.
Calculation formula is introduced in
Explanation 4-1.
Determination Coef. Used for evaluating the linearity of calibration
curve. When this value is closer to 1.0000,
the linearity can be evaluated to be higher.
Calculation formula is introduced in
Explanation 4-1.

4 - 27
4.3 Data Processing

Explanation 4-1 Calculation Formulae for Concentration Difference, Correlation

Coefficient, Determination Coefficient, etc. of Calibration Curve

Introduced below are calculation formulae for evaluating a calibration curve.

An : Absorbance value obtained by

measuring standard samples
A3 A : Averaged absorbance value
obtained by measuring standard
A2 samples
Absorbance

Cstdn : Concentration of each standard

Cn : Concentration on calibration curve,
which corresponds to An
C : Averaged concentration on
calibration curve, which
A1 corresponds to An
Cstd1 Cstd2 Cstd3 N : Number of standards
C1 C2 C3

Concentration

Measured value and calibration curve

Concentration difference DIFF : DIFFn = C n − C stdn

DIFFn
Relative difference RD : RDn = × 100
A
DIFFn
Student’s t (t-test) : tn =
∑ ( DIFF ) n
2

N −1

Relative coefficient : R=
(∑ (C n ) )− (∑ (C
−C
2
n − C stdn )
2
)
∑ (C − C )
2
n

R 2 = (R )
2
Determination coefficient :

4 - 28
 4.3.3

4.3.3 Deletion and Recovery of Calibration Curve Data

1. Displaying Calibration Curve Data

[For check from saved data]

For check by opening the saved data, open the photometric

data including the saved calibration curve data according to
section 4.1.1.
Upon opening the calibration curve data, a guidance (an
example shown in Fig. 4-29) is presented. Select the <2>
key (for Curve).

Select function. :_
[1]Meas. [2]Curve [3]Save [4]Print
◄►( 1/ 1)

Fig. 4-32 Example of Guidance upon

Opening of Calibration Curve Data

[During measurement]

During measurement by selecting Photometry in the Main

Menu, Conc for Data Mode and 1st Order for Curve Type,
select Curve Confirmation or Curve indicated in the
guidance.

2. Displaying Standard Sample Data

(1) The calibration curve data is now opened. Select the

<3> key (for STD).

Photometry/Curve CELL A
R = 0.9999 R2 = 0.9999
+
0.893 +
+
+
ABS +
+
+
0.001 + +
0.000 CONC 90.000

Select function. :_
[1]Scale [2]Factor [3]STD

Fig. 4-33 Calibration Curve Data

4 - 29
4.3 Data Processing

(2) Each standard data is now displayed as shown below.

Photometry/Curve CELL A
ID ABS CONC
STD1 0.001 0.000
STD2 0.099 10.00
STD3 0.203 20.00
STD4 0.300 30.00
STD5 0.405 40.00

Select function. :_
[1] Remeasure [2] Delete [3] Recover

Fig. 4-34 Each Standard Data

Table 4-14 Guidance for Photometry/Curve

Function Key Explanation

[1] Remeasure To be selected for remeasuring an already
measured sample.
Upon selection, the following guidance
appears. Select a sample to be
remeasured by the [▲]/[▼] key and place
the sample in cell 1. Then, press the [Start]
key. Remeasurement will start. For auto
zero operation, press the [AUTOZERO] key.
Photometry CELL A
ID ABS CONC
6 0.301 30.00
7 0.251 25.11
8 0.419 40.90
9 0.110 11.00
10 0.234 23.50

Set sample to Cell1(Sample 10).: _

START:Meas.; AUTOZERO:Autozero
◄►( 2/ 2)

[2] Delete The measured data of a standard which you

don’t want to use for a calibration curve can
be excluded from calculation by selecting the
data by [▲]/[▼] key and pressing the <1>
key (for Delete).
After deletion, the asterisk (*) mark is put at
the head of the data.
Select function. :_
[1]Delete

[3] Recover The measured data of a standard which has

been excluded from calculation can be
recovered by selecting the data by [▲]/[▼]
key and pressing the <1> key (for Recover).
After recovery, the asterisk (*) mark put at
the head of the data will disappear.
Select function. :_
[1] Recover

4 - 30
 4.3.4

4.3.4 Scale Change (Wavelength/Time Scan Data)

1. Displaying Data

[For check from saved data]

For check by opening the saved data, open the saved

wavelength scan data or time scan data according to
section 4.1.1 .

[During measurement]

When measurement is in progress with WL Scan or Time

Scan selected in Main Menu, you should wait till the end of
the measurement.

2. Data Processing Screen

For wavelength scan data, the WL Scan screen (Fig. 4-35)

appears, and the Time Scan screen (Fig. 4-36) appears for
time scan data. Then, select the <2> key (for Scale).

WL Scan CELL A

0.800

ABS

0.000

400.0 525.0 650.0 nm

START:Meas.; Select function.：_

[1]Data [2]Scale [3]Save [4]Print [5]Baseline

Fig. 4-35 Wavelength Scan Data

Time Scan CELL A

0.800

ABS

0.000
0 60.0 120s

START:Meas.; Select function.：_

[1]Data [2]Scale [3]Save [4]Print

Fig. 4-36 Time Scan Data

4 - 31
4.3 Data Processing

3. Changing the Scale

For wavelength scan data, the scale change screen for

wavelength scan (Fig. 4-37) will appear. For time scan
data, the scale change screen for time scan (Fig. 4-38) will
appear. On each screen, scale change, scale resetting
and auto scale operation can be set. For each parameter,
refer to Table 4-12 which lists scale change parameters.

WL Scan/Scale CELL A
Scale Change
X-Max(nm) 650.0
X-Min(nm) 400.0
Y-Max(ABS) 0.800
Y-Min(ABS) 0.000

Scale Reset
Auto Scale

Input value. ： ＿
X-Max(nm) [191.0 – 1100.0]

Fig. 4-37 Scale Changing Screen for

Wavelength Scan

Time Scan/Scale CELL A

Scale Change
X-Max (s) 60
X-Min (s) 0
Y-Max (ABS) 9.999
Y-Min (ABS) 0.000

Scale Reset
Auto Scale

Input value. ： ＿
X-Max (s) [10 - 99999]

Fig. 4-38 Scale Changing Screen for Time Scan

4 - 32
 4.3.4

Table 4-15 Scale Changing Parameters for

Wavelength Scan

Function Key Explanation

X-Max Sets the maximum value on the X axis (wavelength
axis) of spectrum.
Settable within 191.0 to 1100.0. For adopting and
changing the set axis, return to the spectrum screen
by pressing the [RETURN] key.
X-Min Sets the minimum value on the X axis (wavelength
axis) of spectrum.
Settable within 190.0 to 1099.0. For adopting and
changing the set axis, return to the spectrum screen
by pressing the [RETURN] key.
Y-Max Sets the maximum value on the Y axis (representing
the absorbance or transmittance) of spectrum.
Settable range is as follows.
Absorbance : -9.999 to 9.999
Transmittance : -9.999 to 999.9
For adopting and changing the set axis, return to the
spectrum screen by pressing the [RETURN] key.
Y-Min Sets the minimum value on the Y axis (representing
the absorbance or transmittance) of spectrum.
Settable range is as follows.
Absorbance : 9.999 to 9.999
Transmittance : -9.999 to 999.9
For adopting and changing the set axis, return to the
spectrum screen by pressing the [RETURN] key.
Scale Reset Sets scale in the default value. For setting, press
the <1> key (Execute).
Select function. :_
[1]Execute

Auto Scale Judges the maximum and minimum values of the

measured data on the Y axis, and automatically
optimizes the scale. For setting, press the <1> key
(Execute).
Select function. :_
[1]Execute

4 - 33
4.3 Data Processing

Table 4-16 Scale Changing Parameters for Time Scan

Function Key Explanation

X-Max Sets the maximum value on the X axis (time axis) of
spectrum.
Settable within 10 to 99999. For adopting and
changing the set axis, return to the spectrum screen
by pressing the [RETURN] key.
X-Min Sets the minimum value on the X axis (time axis) of
spectrum.
Settable within 0 to 99989. For adopting and
changing the set axis, return to the spectrum screen
by pressing the [RETURN] key.
Y-Max Sets the maximum value on the Y axis (representing
the absorbance) of calibration curve.
Settable range is as follows.
Absorbance : -9.999 to 9.999
Transmittance : -9.999 to 999.9
For adopting and changing the set axis, return to the
spectrum screen by pressing the [RETURN] key.
Y-Min Sets the minimum value on the Y axis (representing
the absorbance) of spectrum.
Settable range is as follows.
Absorbance : -9.999 to 9.999
Transmittance : -9.999 to 999.9
For adopting and changing the set axis, return to the
spectrum screen by pressing the [RETURN] key.
Scale Reset Sets scale in the default value. For setting, press
the <1> key (Execute).
Select function. :_
[1]Execute

Auto Scale Judges the maximum and minimum values of the

measured data on the Y axis, and automatically
optimizes the scale. For setting, press the <1> key
(Execute).
Select function. :_
[1]Execute

4 - 34
 4.3.5

4.3.5 Peak Detection (Wavelength Scan Data)

1. Displaying Data

[For check from saved data]

For check by opening the saved data, open the saved

wavelength scan data according to section 4.1.1.

[During measurement]

When measurement is in progress with WL Scan selected

in Main Menu, you should wait till the end of the
measurement.

2. Data Processing Screen

(1) The guidance for wavelength scan data (Fig. 4-39) is

then opened. In this guidance, select the <1> key (for
Data). The guidance for data screen (Fig. 4-40) will
be displayed. In this guidance, select the <2> key (for
Peak).

WL Scan CELL A

0.800

ABS

0.000
400.0 525.0 650.0 nm

START:Meas.; Select function.：_

[1]Data [2]Scale [3]Save [4]Print [5]Baseline

Fig. 4-39 Guidance for Wavelength Scan Data

Select function. :_
[1]Trace [2]Peak [3]Smoothing [4]List

Fig. 4-40 Guidance for Data Screen

4 - 35
4.3 Data Processing

3. Peak Detection Screen

(1) The peak detection screen (Fig. 4-41) is now opened.

The peak and valley detected according to the peak
parameters set for measurement are indicated by +
marks at the peak positions on the spectral profile.
When plural peaks and valleys have been detected,
you can move to different peak/valley by pressing the
[◄]/[►] key. The wavelength and photometric value
at the cursor position are indicated at the top left of this
screen. Also, the present peak detection parameters
are indicated at the right of the wavelength and
photometric value. For changing the peak/valley
detection parameters, select the <1> key (for Peak
PARAM.), and the <2> key (for Peak Table) for
displaying the peak table. For details of Peak PARAM.
and Peak Table, refer to Table 4-17.

WL Scan CELL A
546.0 nm 0.286 ABS (T = 0.010, S = 2)
0.367
＋
＋
ABS
＋ ＋
＋
＋ ＋
0.278
400.0 500.0 600.0 nm

Select function. :_
[1]Peak PARAM. [2]Peak Table

Fig. 4-41 Peak Detection Screen

4 - 36
 4.3.5

Table 4-17 Peak Detection Data Processing

Function Key Explanation

[1] Peak PARAM. Used for changing the peak and valley
detection parameters. For details of each
parameter, refer to Table 3-39 which lists
calculation parameters.
WL Scan/Peak CELL A
Peak PARAM.
Threshold 0.010
Sensitivity 2

Input value. ： ＿
Threshold [0.001 – 1.000]

[2] Peak Table Displays peak table. Each wavelength of

the detected peak and valley, and the
photometric value at the wavelength are
indicated. Peak and valley data are
indicated at the left and right, respectively.
WL Scan/Peak CELL A
Peak Table (T=0.010 S=2)
No WL(nm) Peak WL(nm) Valley
1 498.0 0.297 546.0 0.286
2 468.0 0.290

Select function. :_
1 Peak PARAM. □
□ 2 Peak Trace
◄►( 1/ 1)

4 - 37
4.3 Data Processing

4.3.6 Data Tracing (Wavelength/Time Scan Data)

1. Displaying Data

[For check from saved data]

For check by opening the saved data, open the saved

wavelength scan data or time scan data according to
section 4.1.1..

[During measurement]

When measurement is in progress with WL Scan or Time

Scan selected in Main Menu, you should wait till the end of
the measurement. For stopping the measurement, press
the [STOP] key.

2. Data Processing Screen

(1) For wavelength scan data, the WL Scan screen (Fig. 4-

42) appears, and the Time Scan screen (Fig. 4-43)
appears for time scan data. Then, select the <1> key
(for Data).

WL Scan CELL A
0.800

ABS

0.000
400.0 525.0 650.0 nm

START:Meas.; Select function.：_

[1]Data [2]Scale [3]Save [4]Print [5]Baseline

Fig. 4-42 Wavelength Scan Data

Time Scan CELL A

0.800

ABS

0.000
0 60.0 120 s
START:Meas.; Select function.：_
[1]Data [2]Scale [3]Save [4]Print

Fig. 4-43 Time Scan Data

4 - 38
 4.3.6

(2) For wavelength scan data, the guidance for

wavelength scan (Fig. 4-44) is displayed. And, for
time scan data, the guidance for time scan (Fig. 4-45)
is displayed. On each screen, select the <1> key (for
Trace).

Select function. :_
[1]Trace [2]Peak [3]Smoothing [4]List

Fig. 4-44 Guidance for Wavelength Scan

Select function. :_
[1]Trace [2]Smoothing [3]List

Fig. 4-45 Guidance for Time Scan

3. Tracing Data

For wavelength scan data, the WL Scan/Trace screen (Fig.

4-46) is displayed. And, for time scan data, the Time
Scan/Trace screen (Fig. 4-47) is displayed. The cursor on
each screen can be moved by pressing the [◄]/[►] key.
The wavelength and photometric value at the present
cursor position are indicated at the top left of each screen.
Thus, the photometric values at the aimed-at wavelength
and time point can be read.

WL Scan/Trace CELL A
523.0 nm 0.650 ABS
0.800
＋
ABS

0.000
400.0 525.0 650.0 nm

◄►

Fig. 4-46 WL Scan/Trace Screen

4 - 39
4.3 Data Processing

Time Scan/Trace CELL A

75 sec 0.402 ABS
0.800

ABS
＋
0.000
0 60.0 120s

◄►

Fig. 4-47 Time Scan/Trace Screen

4 - 40
 4.3.7

4.3.7 Data Smoothing (Wavelength/Time Scan Data)

1. Displaying Data

[For check from saved data]

For check by opening the saved data, open the saved

wavelength scan data or time scan data according to
section 4.1.1..

[During measurement]

When measurement is in progress with WL Scan or Time

Scan selected in Main Menu, you should wait till the end of
the measurement. For stopping the measurement, press
the [STOP] key.

2. Data Processing Screen

(1) For wavelength scan data, the WL Scan screen (Fig. 4-

48) appears, and the Time Scan screen (Fig. 4-49)
appears for time scan data. Then, select the <1> key
(for Data).

WL Scan CELL A

0.800

ABS

0.000
400.0 525.0 650.0 nm

START:Meas.; Select function.：_

[1]Data [2]Scale [3]Save [4]Print [5]Baseline

Fig. 4-48 Wavelength Scan Data

4 - 41
4.3 Data Processing

Time Scan CELL A

0.800

ABS

0.000
0 60.0 120 s

START:Meas.; Select function.：_

[1]Data [2]Scale [3]Save [4]Print

Fig. 4-49 Time Scan Data

(2) For wavelength scan data, the guidance for

wavelength scan (Fig. 4-50) is displayed. In this
guidance, select the <3> key (for Smoothing).
For time scan data, the guidance for time scan (Fig. 4-
51) is displayed. In this guidance, select the <2> key
(for Smoothing).

Select function. :_
[1]Trace [2]Peak [3]Smoothing [4]List

Fig. 4-50 Guidance for Wavelength Scan

Select function. :_
[1]Trace [2]Smoothing [3]List

Fig. 4-51 Guidance for Time Scan

3. Smoothing Data

For wavelength scan data, the WL Scan/Smoothing screen

(Fig. 4-52) is displayed. And, for time scan data, the Time
Scan/Smoothing (Fig. 4-53) is displayed. For smoothing,
select the <1> key (for Smoothing) on each screen.
Upon selection, the spectral profile on the screen is
smoothed. The spectral profile can be smoothed
repeatedly by selecting the <1> key (for Smoothing)
repeatedly. For returning to the original data, select the
<2> key (for Reset).

4 - 42
 4.3.7

WL Scan/Smoothing CELL A

0.800

ABS

0.000
400.0 525.0 650.0 nm

Select function. :_
[1]Smoothing [2]Reset

Fig. 4-52 WL Scan/Smoothing Screen

Time Scan/Smoothing CELL A

0.800

ABS

0.000
0 60.0 120 s

Select function. :_
[1]Smoothing [2]Reset

Fig. 4-53 Time Scan/Smoothing Screen

4 - 43
4.3 Data Processing

4.3.8 Displaying Data in List (Wavelength/Time Scan Data)

1. Displaying Data

[For check from saved data]

For check by opening the saved data, open the saved

wavelength scan data or time scan data according to
section 4.1.1..

[During measurement]

When measurement is in progress with WL Scan or Time

Scan selected in Main Menu, you should wait till the end of
the measurement. For stopping the measurement, press
the [STOP] key.

2. Data Processing Screen

(1) For wavelength scan data, the WL Scan screen (Fig. 4-

54) appears, and the Time Scan screen (Fig. 4-55)
appears for time scan data. Then, select the <1> key
(for Data).

WL Scan CELL A
0.800

ABS

0.000
400.0 525.0 650.0 nm
START:Meas.; Select function.：_
[1]Data [2]Scale [3]Save [4]Print [5]Baseline

Fig. 4-54 Wavelength Scan Data

Time Scan CELL A

0.800

ABS

0.000
0 60.0 120 s
START:Meas.; Select function.：_
[1]Data [2]Scale [3]Save [4]Print

Fig. 4-55 Time Scan Data

4 - 44
 4.3.8

(2) For wavelength scan data, the guidance for

wavelength scan (Fig. 4-56) is displayed. In this
guidance, select the <4> key (for List). For time scan
data, the guidance for time scan (Fig. 4-57) is
displayed. In this guidance, select the <3> key (for
List).

Select function. :_
[1]Trace [2]Peak [3]Smoothing [4]List

Fig. 4-56 Guidance for Wavelength Scan

Select function. :_
[1]Trace [2]Smoothing [3]List

Fig. 4-57 Guidance for Time Scan

3. Displaying data in list

For wavelength scan data, the WL Scan/List screen (Fig. 4-

58) is displayed. And, for time scan data, the Time
Scan/List screen (Fig. 4-59) is displayed. You can see the
next or previous page on each screen by pressing the
[◄]/[►] key. For changing the display interval, select the
<1> key (for Int. (nm/s)). For details of setting method,
refer to the data list screen (shown in Table 4-18).

WL Scan/List CELL A
No. WL(nm) ABS No. WL(nm) ABS
1 600.0 0.299 9 584.0 0.298
2 598.0 0.299 10 582.0 0.297
3 596.0 0.299 11 580.0 0.296
4 594.0 0.299 12 578.0 0.295
5 592.0 0.299 13 576.0 0.294
6 590.0 0.299 14 574.0 0.293
7 588.0 0.299 15 572.0 0.292
8 586.0 0.299 16 570.0 0.291
Select function. :_
[1]Int.(nm) (2.0)
◄►( 1/ 7)

Fig. 4-58 WL Scan/List Screen

4 - 45
4.3 Data Processing

Time Scan/List CELL A

No. TIME(s) ABS No. TIME(s) ABS
1 0 0.000 9 8 0.001
2 1 0.001 10 9 0.002
3 2 0.000 11 10 0.003
4 3 0.000 12 11 0.007
5 4 0.001 13 12 0.013
6 5 0.000 14 13 0.020
7 6 0.000 15 14 0.034
8 7 0.000 16 15 0.055

Select function. :_
[1]Int.(nm) (1)
◄►( 1/ 3)

Fig. 4-59 Time Scan/List Screen

Table 4-18 Data List Screen

Function Key Explanation

[1] Int. (nm/s) To be selected for changing the display interval.
Input a display interval value, and press the
[RETURN] key for returning to each initial data list
screen.
WL Scan/List CELL A
Int.(nm) 1.0

Input value. ： ＿
Int.(nm) [0.1 – 100.0]

Time Scan/List CELL A

Int.(s) 1

Input value. ： ＿
Int.(s) [1 – 100.0]

4 - 46
 4.4

4.4 Measurement in Statistical Calculation Mode

In quantitation of solution sample concentration and in

measurement of a DNA sample (by ratio calculation), the mean
value (MEAN), standard deviation (SD) and relative standard
deviation (RSD) can be calculated every specified number of
calculations when you make setting for statistical calculation
before measurement.

GUIDE: When auto start has been set, parameters are

automatically set after the power switch is turned on
and the process advances up to the measurement
screen. For automatic start setting method, see
section 4.5..

[For quantitation of solution sample concentration]

Depending on each mode, set measurement parameters by

reference to the following subsections. However, in setting of
sample conditions, you must set the statistical calculation in ON
status, and set the number of samples for statistical calculation
in setting the number of calculations.

6-cell auto mode:

3.2.1 Determining Concentration of Solution
6-cell manual mode, 6-cell mode OFF:
3.3.1 Determining Concentration of Solution

After measurement, the result of statistical calculation is

displayed every specified number of calculations as shown
below. N, MEAN, SD and RSD stand for the number of
calculations, mean concentration value, standard deviation in
concentration and relative standard deviation in concentration,
respectively.

Photometry CELL A
ID ABS CONC
1 0.098 9.980
2 0.099 9.990
N=2 MEAN = 9.985, SD = 0.007, RSD = 0.1
3 0.055 5.500
4 0.053 5.300
N=2 MEAN = 5.400, SD = 0.141, RSD = 2.6
5 0.098 9.980
6 0.099 9.990

Fig. 4-60 Result of Statistical Calculation in

Quantitation of Solution Sample Concentration

4 - 47
4.4 Measurement in Statistical Calculation Mode

[For measurement of DNA sample (by ratio calculation)]

Depending on each mode, set measurement parameters by

reference to the following subsections. However, in setting of
sample conditions, you must set the statistical calculation in ON
status, and set the number of samples for statistical calculation
in setting the number of calculations.

6-cell auto mode:

3.2.3 Measuring DNA Sample (measurement through ratio
calculation)
6-cell manual mode, 6-cell mode OFF:
3.3.3 Measuring DNA Sample (measurement through ratio
calculation)

After measurement, the result of statistical calculation is

displayed every specified number of calculations as shown
below. N, MEAN, SD and RSD stand for the number of
calculations, mean absorbance difference or ratio value,
standard deviation and relative standard deviation, respectively.

Photometry 280.0 nm 0.268 ABS CELL A

ID ABS
2 WL1 （ 260.0 nm） 0.423
WL2 （ 280.0 nm） 0.268
N=2 [MEAN] [SD] [RSD]
ABS DIFF = 0.155 [0.155] [0.003] [0.1]
ABS Ratio= 1.582 [1.582] [0.004] [0.2]

Fig. 4-61 Result of Statistical Calculation in

DNA Measurement

4 - 48
 4.5

4.5 Auto Start Function

The instrument can always be started up under the same

measurement parameters by using the auto start function.
This is a very useful function when measurement is always
carried out in only one mode.

1. Startup of This Product

Start up this product. (For startup method, see section

2.3.1..)

2. Method Loading Procedure

(1) Press the [MAIN MENU] key. The Main Menu screen
(shown in Fig. 4-62) appears.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. :_

Fig. 4-62 Main Menu Screen

(2) For loading a method, press the <4> key (for Method
Menu) and then the [ENTER] key or press the [▲]/[▼]
key to select Method Menu and then the [ENTER] key.

(3) Method Menu screen (Fig. 4-63) appears. For auto

start setting, press the <2> key or the [▲]/[▼] key to
select Auto Start, and then the [ENTER] key.

Meas.PARAM. CELL A
1 Load/Delete Method
2 Auto Start
3 Print Method List

Select function. :_

Fig. 4-63 Meas. PARAM. Screen

4 - 49
4.5 Auto Start Function

3. Auto Start Setting

(1) The Auto Start screen (Fig. 4-64) appears. On this

screen, the saved methods are listed. On each row,
file name, type (PHT: photometry, WLS: wavelength
scan, TMS: time scan) and date of file saving are
shown from the left. Select the method to be
automatically started by the [▲]/[▼] key and press the
[ENTER] key. For the set method, an asterisk (*) is
put at the head of its file name.
Figure 4-65 shows the screen when the file name “1
NO3-(Hitachi Taro)” is selected for auto start.

Auto Start CELL A

1 NO3-(Hitachi Taro) PHT 2009/10/10
2 Cr6+(Hitachi Hanako) PHT 2009/12/01
3 Spectrum WLS 2009/12/10
4 Time Scan TMS 2010/01/02
5 Fe PHT 2010/01/12
6 DNA PHT 2010/02/05
7 Stability TMS 2010/03/10

Type Date of
File name
Select function. :_
saving
[1]Confirmation [2]Type [3]Sort
◄ ► ( 1/ 1)

Fig. 4-64 Auto Start Screen

Auto Start CELL A

*1 NO3-(Hitachi Taro) PHT 2009/10/10
2 Cr6+(Hitachi Hanako) PHT 2009/12/01
3 Spectrum WLS 2009/12/10
4 Time Scan TMS 2010/01/02
5 Fe PHT 2010/01/12
6 DNA PHT 2010/02/05
7 Stability TMS 2010/03/10

Select function. :_
[1]Confirmation [2]Type [3]Sort
◄ ► ( 1/ 1)

Fig. 4-65 Auto Start-Set Status

Same Method is always set when the device is started

up by doing the above-mentioned setting, and it shifts
to the measurement screen.

Note ：
Please confirm whether the same attachment device
as Method File that does Auto Start without fail has
been installed before the device is started up. For
instance, please confirm 6-Cell holder is installed in the

4 - 50
 4.5

device when Method File that does Auto Start is a file

using 6-Cell holder. Moreover, please confirm the
same attachment device is installed for Method using
the single cell folder (another sales) and the square
length cell folders (another sales). Please confirm
whether Auto Sipper (another sales) is installed in the
device for Method using Auto Sipper (another sales).

For releasing the auto start setting, select the file for
which auto start has been set, and then press the
[ENTER] key. Auto start is now canceled.
Make sure the asterisk has disappeared.

4 - 51
4.6 Introduction and Mounting Method of Separately Available Options

4.6 Introduction and Mounting Method of Separately Available Options

Introduced below are separately available options.

These options should be used according to your analytical
purpose. Of the single cell holders, mask for micro cell and
rectangular long-path cell holder, mounting method and
instrument setting method are explained in 4.6.1 to 4.6.5.

Table 4-19 Introduction of Separately Available Options

Sample
Purpose Option (separately available)
Volume
Measurement Single cell P/N: 3J2-0110 1.7 to 3.5 mL
with usual cell holder
holder
Application to Single cell P/N: 3J2-0110 340 to 600 μL
small-volume holder
samples Mask for micro P/N: 200-1537
cell
10 mm quartz P/N: 124-0357
micro cell
10 mm black P/N: 200-0551
micro cell
Sensitivity Rectangular P/N: 3J2-0111 17 to 35 mL
enhancement long-path cell
holder
10 mm quartz P/N: 210-3939
cell
Use of sipper Auto sipper P/N: 3J2-0100 ⎯

For these separately available options and the hottest

information about them, contact your dealer or the maintenance
service company authorized by Hitachi High-Technologies
Corporation.

4 - 52
 4.6.1

4.6.1 Single Cell Holder (Separately Available)

A single cell holder has only one cell setting position like the
conventional cell holders. This holder should be employed
when you want to use cells by the usual method.

1. Removal of 6-cell Holder

Remove the 6-cell holder, referring to the 6-cell holder

removing method described in “a.” of 2.4.5.

2. Mounting of Single Cell Holder

(1) Open the sample compartment and place a single cell

holder so that its positioning holes receive the guide
pins of the sample compartment. The single cell
holder must be mounted facing the rectangular cell
setting position toward the front of the instrument as
shown below.

Guide pins

Fig. 4-66 Guide Pins of Sample Compartment

Positioning holes

Rectangular cell
setting position

Face toward instrument front.

Fig. 4-67 Appearance of Single Cell Holder

4 - 53
4.6 Introduction and Mounting Method of Separately Available Options

(2) Only one screw is used for fastening both the sample
compartment and single cell holder. Tighten the
screw firmly using a Phillips screwdriver.

Screw

Fig. 4-68 Mounting Single Cell Holder in Sample

Compartment

(3) Holder mounting is now completed. Close the lid of

the sample compartment.

Fig. 4-69 View After Mounting the Single Cell Holder

3. Change of 6-cell Mode Setting

The 6-cell mode setting needs to be changed.

Turn the 6-cell mode setting from ON to OFF, referring to
section 2.5.4.
Carry out measurement, after referring to section 4.6.5.

4 - 54
 4.6.2

4.6.2 Mask for Micro Cell (Separately Available)

The mask for micro cell is used in combination with a single cell
holder and a micro cell. In this combination, a sample is
measurable in a smaller volume (340 to 600 μL) than with a 10
mm rectangular cell.

1. Mounting of Single Cell Holder

Mount a single cell holder in the same procedure as in 4.6.1.

2. Mounting of Mask for Micro Cell

(1) Shown below are the external view of this mask and its
illustration drawn as viewed in the lateral direction.
The swelled side of the mask is shown as the “Convex
side of mask” and the sunk side is shown as the
“Concave side of mask” in the figure.

Mask face

Convex side Concave side

of mask of mask

Fig. 4-70 External View and Illustration of Mask for

Micro Cell

(2) Shown below is the external view of a single cell

holder. The cell holder has a groove for mask
insertion. Insert the mask into the groove by sliding.
The mask must be inserted orienting the convex side
as directed in the figure. Insert the mask completely
until it no longer moves.

4 - 55
4.6 Introduction and Mounting Method of Separately Available Options

Concave side of
mask Groove for inserting
mask for micro cell

Orient the convex side

of mask on this side.

Fig. 4-71 External View of Single Cell Holder

Insert the mask for micro cell

into the groove by sliding.
The mask must be inserted
completely until it no longer
moves.

Fig. 4-72 Insertion of Mask for Micro Cell

3. Change of 6-cell Mode Setting

The 6-cell mode setting needs to be changed.

Turn the 6-cell mode setting from ON to OFF, referring to
section 2.5.4.
Carry out measurement, after referring to section 4.6.5.

4 - 56
 4.6.3

4.6.3 Rectangular Long-Path Cell Holder (Separately Available)

The rectangular long-path cell holder enables the user to

measure a sample using a cell having an optical path length
within 10 to 100 mm. In this application, low-concentration
samples can be measured at enhanced absorbance levels.

1. Removal of 6-cell Holder

Remove the 6-cell holder, referring to the 6-cell holder

removing method described in “a.” of 2.4.5.

2. Mounting of Rectangular Long-path Cell Holder

(1) Open the sample compartment and place the
rectangular long-path cell holder so that its positioning
holes receive the guide pins of the sample
compartment. This holder must be mounted locating
the rectangular cell mounting position on the front side
of the instrument as shown below.

Guide pins

Fig. 4-73 Guide Pins of Sample Compartment

Positioning holes

Locate on the front side

of instrument.

Fig. 4-74 Appearance of Rectangular Long-Path

Cell Holder

4 - 57
4.6 Introduction and Mounting Method of Separately Available Options

(2) Only one screw is used for fastening both the sample
compartment and rectangular long-path cell holder.
Tighten the screw firmly using a Phillips screwdriver.

Screw

Fig. 4-75 Mounting Rectangular Long-Path

Cell Holder in Sample Compartment

(3) Holder mounting is now completed. Close the lid of

the sample compartment.

Fig. 4-76 View After Mounting the Rectangular

Long-Path Cell Holder

3. Change of 6-cell Mode Setting

The 6-cell mode setting needs to be changed.

Turn the 6-cell mode setting from ON to OFF, referring to
section 2.5.4.
Carry out measurement, after referring to section 4.6.5.

4 - 58
 4.6.4

4.6.4 How to Return 6-Cell Holder

1. Removal of Present Cell Holder

Firstly, remove the mask for micro cell when mounted to the
single cell holder. Then, remove the single cell holder or
rectangular long-path cell holder, whichever mounted, by
loosening the screw shown in the figure below.

Screw

Fig. 4-77 Removal of Single Cell Holder

Screw

Fig. 4-78 Removal of Rectangular Long-Path Cell Holder

2. Mounting of 6-cell Holder

Mount the 6-cell holder, referring to the 6-cell holder

mounting method described in “b.” of 2.4.5.

3. Change of 6-cell Mode Setting

The 6-cell mode setting needs to be changed.

Turn the 6-cell mode setting from OFF to ON, referring to
section 2.5.4.

4 - 59
4.6.5 Measurement in 6-Cell Mode OFF Status

Explained here is the measurement method when the 6-cell

mode is set at OFF for using a single cell holder or rectangular
long-path cell holder. When the 6-cell mode is set at OFF,
measurement should be carried out, referring to section 3.3
except for the following.

(1) Because 6-cell mode OFF is a method to be used when the

6-cell holder is not used (i.e., a single cell holder or
rectangular long-path cell holder is used), operations
involving a movement of the 6-cell holder cannot be
performed.

(2) For execution of auto zero, a guidance “Set autozero

sample to Cell A.” is issued in the 6-cell manual mode.
However, in the 6-cell mode OFF status, cell position is not
specified (by omitting “to Cell A”). So, set the sample in
the cell holder in the sample compartment.

(3) For baseline correction in spectrum measurement, a

guidance “Set baseline sample to Cell A.” is issued in the 6-
cell manual mode. However, in the 6-cell mode OFF
status, cell position is not specified (by omitting “to Cell A”).
So, set the sample in the cell holder in the sample
compartment.

(4) In the guidance for cell position (see section 2.4.2), Cell S
is always indicated when the 6-cell mode is set at OFF.

(5) For execution of auto zero in measurement via the enlarge

display screen (Data Display), firstly place a sample for
auto zero execution in the cell holder, and then press the
[Autozero] key. Upon pressing this key, auto zero is
directly executed.

4 - 60
 5.1

5. PERFORMANCE CHECK

This section pertains to the performance check method for

checking whether the instrument provides the specified
performance. For performance check with a pen type low-
pressure mercury lamp, a separately available option is required.
For mounting method of this lamp, refer to the instruction manual
for the separately available option.

5.1 Check with Instrument Main Unit Alone

(1) After startup of the instrument, press the [MAIN MENU] key
to display the Main Menu screen (Fig. 5-1). Confirm that
nothing is contained in the sample compartment and close
the lid. For selection of performance check, press the <7>
key (Performance) and then the [ENTER] key or press the
[▲]/[▼] key to select Performance followed by the [ENTER]
key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 5-1 Main Menu Screen

(2) The instrument performance check screen (Fig. 5-2)

appears.

Performance CELL A
1 Performance Check(Auto)
2 Performance Check(Manual)
3 Performance Check(Option)

Select function. : _

Fig. 5-2 Instrument Performance Check Screen

5-1
5.1 Check with Instrument Main Unit Alone

[For automatic execution of instrument performance check]

For automatically executing instrument performance check,

press the <1> key (Performance Check (Auto)) and then
the [ENTER] key. For subsequent operation, refer to
section 5.1.8.

[For instrument performance check in each item]

For checking instrument performance in each performance

item, press the <2> key (Performance Check (Manual)) and
then the [ENTER] key. For subsequent operation, refer to
section 5.1.1 to 5.1.6.

[For instrument performance check with separately

available Hg lamp]

For checking instrument performance with a Hg lamp

available at option, press the <3> key (Performance Check
(Option)) and then the [ENTER] key. For subsequent
operation, refer to section 5.2.1 to 5.2.3.”

5-2
 5.1.1

5.1.1 Wavelength Accuracy

(1) Display the screen for each item of instrument performance

check (Fig. 5-3). For display method, follow the procedure
starting from section 5.1.. Using the [▲]/[▼] key, move
the cursor to the measuring wavelength in any of WL
Accuracy (484.6nm), (229.0nm) and (822.8nm), and then
select [1] Yes. The accuracy at the selected wavelength
will be checked. If check is not desired actually, select [2]
No.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
[1]Yes [2]No

Fig. 5-3 Screen for Each Item of Instrument

Performance Check

(2) The guidance for informing the parameter setting process

(Fig. 5-4) appears, and parameters will be set and
wavelength accuracy will be checked.

Setting PARAM....

Fig. 5-4 Guidance for Parameter Setting Process

(3) After the check of wavelength accuracy, the screen after

this check is displayed as shown in Fig. 5-5.
Measurement result is indicated on the wavelength
accuracy line; a peak difference at each wavelength of
bright line and OK when the judgment standard is met or
NG if not met. If NG is indicated, carry out wavelength
calibration by the method instructed in section 5.3.1, and
then check the wavelength accuracy again in the correct
procedure. If NG recurs, refer to section 6.8.

5-3
5.1 Check with Instrument Main Unit Alone

Performance CELL A
WL Accuracy (484.6 nm) -0.2nm OK
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Select function. : _
[1]Print

Fig. 5-5 Screen after Wavelength Accuracy Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-1 Measuring Conditions and Judgment Standard

for Wavelength Accuracy (at 484.6 nm)

Item Wavelength Accuracy (at 484.6 nm)

Measuring Emission spectrum of Xe lamp bright line
conditions (with detector on the monitor side)
Wavelength range : 478.6 to 490.6 nm
Scan speed : 40 nm/min
Data interval : High resolution
Response : Standard
Spectrum to be measured
Calculation method Determination of a difference between
peak wavelength of the measured
spectrum and 484.6 nm
Wavelength accuracy (at 484.6 nm) =
(obtained peak wavelength) - 484.6
Judgment standard Within ±1.0 nm

5-4
 5.1.1

Table 5-2 Measuring Conditions and Judgment Standard

for Wavelength Accuracy (at 229.0 nm)

Item Wavelength Accuracy (at 229.0 nm)

Measuring Emission spectrum of Xe lamp bright line
conditions (with detector on the monitor side)
Wavelength range : 223.0 to 235.0 nm
Scan speed : 40 nm/min
Data interval : High resolution
Response : Standard
Spectrum to be measured
Calculation method Determination of a difference between
peak wavelength of the measured
spectrum and 229.0 nm
Wavelength accuracy (at 229.0 nm) =
(obtained peak wavelength) - 229.0
Judgment standard Within ±2.0 nm

Table 5-3 Measuring Conditions and Judgment Standard

for Wavelength Accuracy (at 822.8 nm)

Item Wavelength Accuracy (at 822.8 nm)

Measuring Emission spectrum of Xe lamp bright line
conditions (with detector on the monitor side)
Wavelength range : 816.8 to 828.8 nm
Scan speed : 40 nm/min
Data interval : High resolution
Response : Standard
Spectrum to be measured
Calculation method Determination of a difference between
peak wavelength of the measured
spectrum and 822.8 nm
Wavelength accuracy (at 822.8 nm) =
(obtained peak wavelength) - 822.8
Judgment standard Within ±2.0 nm

5-5
5.1 Check with Instrument Main Unit Alone

Fig. 5-6 Example Result of Wavelength Accuracy Check

(at 484.6 nm)

5-6
 5.1.2

5.1.2 Wavelength Repeatability

(1) Display the screen for each item of instrument performance

check (Fig. 5-7). For display method, follow the procedure
starting from section 5.1.. Using the [▲]/[▼] key, move
the cursor to Repeatability of WL, and then select [1] Yes.
Wavelength repeatability will be checked. If check is not
desired actually, select [2] No.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
1 Yes □
□ 2 No

Fig. 5-7 Screen for Each Item of Instrument

Performance Check

(2) The guidance for informing the parameter setting process

(Fig. 5-8) appears, and parameters will be set and
wavelength repeatability will be checked.

Setting PARAM....

Fig. 5-8 Guidance for Parameter Setting Process

(3) After the check of wavelength repeatability, the screen after

this check is displayed as shown in Fig. 5-9.
Measurement result is indicated on the wavelength
repeatability line followed by OK when the judgment
standard is met or NG if not met. If NG is indicated, check
the wavelength repeatability again in the correct procedure.
If NG recurs, refer to section 6.8 .

5-7
5.1 Check with Instrument Main Unit Alone

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability ±0.20 nm OK
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Select function. : _
1 Print
□

Fig. 5-9 Screen after Wavelength Repeatability Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-4 Measuring Conditions and Judgment

Standard for Wavelength Repeatability

Item Wavelength Repeatability

Measuring Emission spectrum of Xe lamp bright line
conditions (with detector on the monitor side)
Wavelength
range : 478.6 to 490.6 nm
Scan speed : 40 nm/min
Data interval : High resolution
Response : Standard
Spectrum to be measured 3 times.
2nd measurement after shifting to
1100 nm. 3rd measurement after shifting
to 190 nm.
Calculation method A difference between maximum and
minimum peak wavelength values in 3
measurements is determined.
Then, wavelength repeatability is
calculated according to the following
formula.
Wavelength repeatability =
±(difference between maximum and
minimum peak wavelength values)/2
Judgment standard Within ±0.5 nm

5-8
 5.1.2

Fig. 5-10 Example Result of Wavelength Repeatability

Check

5-9
5.1 Check with Instrument Main Unit Alone

5.1.3 Noise Level (RMS)

(1) Display the screen for each item of instrument performance

check (Fig. 5-11). For display method, follow the
procedure starting from section 5.1.. Using the [▲]/[▼]
key, move the cursor to Noise Level (RMS), and then select
[1] Yes. Noise level will be checked. If check is not
desired actually, select [2] No.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
1 Yes □
□ 2 No

Fig. 5-11 Screen for Each Item of Instrument

Performance Check

(2) The guidance for informing the parameter setting process

(Fig. 5-12) appears, and parameters will be set and noise
level will be checked.

Setting PARAM....

Fig. 5-12 Guidance for Parameter Setting Process

(3) After the check of noise level, the screen after this check is
displayed as shown in Fig. 5-13. Measurement result is
indicated on the noise level line followed by OK when the
judgment standard is met or NG if not met. If NG is
indicated, check the noise level again in the correct
procedure. If NG recurs, refer to section 6.8.

5 - 10
 5.1.3

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS) 0.0001 ABS OK
Baseline Flatness
Baseline Stability
Hardware
Print Report
Select function. : _
1 Print
□

Fig. 5-13 Screen after Noise Level Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-5 Measuring Conditions and Judgment Standard

for Noise Level (RMS)

Item Noise Level (RMS)

Measuring Time scan measurement (ABS measurement)
conditions Wavelength : 260 nm
Response : Standard
Measurement after auto zero operation
Calculation method Calculation of noise level (RMS) from the
measured absorbance according to the
following formula
n n

∑ ( X i − ∑ ( X i / n)) 2
i =1 i =1
Noise level (RMS) =
n
(n: total number of data points, Xi: ABS at 1st
point)
Judgment standard Within 0.0002 Abs

5 - 11
5.1 Check with Instrument Main Unit Alone

Fig. 5-14 Example Result of Noise Level Check

5 - 12
 5.1.4

5.1.4 Baseline Flatness

(1) Display the screen for each item of instrument performance

check (Fig. 5-15). For display method, follow the
procedure starting from section 5.1.. Using the [▲]/[▼]
key, move the cursor to Baseline Flatness, and then select
[1] Yes. Baseline flatness will be checked. If check is not
desired actually, select [2] No.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
1 Yes □
□ 2 No

Fig. 5-15 Screen for Each Item of Instrument

Performance Check

(2) The guidance for informing the parameter setting process

(Fig. 5-16) appears, and parameters will be set and
baseline flatness will be checked.

Setting PARAM....

Fig. 5-16 Guidance for Parameter Setting Process

(3) After the check of baseline flatness, the screen after this
check is displayed as shown in Fig. 5-17. Measurement
result is indicated on the baseline flatness line followed by
OK when the judgment standard is met or NG if not met. If
NG is indicated, check the baseline flatness again in the
correct procedure. If NG recurs, refer to section 6.8.

5 - 13
5.1 Check with Instrument Main Unit Alone

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness ±0.001 ABS OK
Baseline Stability
Hardware
Print Report
Select function. : _
1 Print
□

Fig. 5-17 Screen after Baseline Flatness Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-6 Measuring Conditions and Judgment

Standard for Baseline Flatness

Item Baseline Flatness

Measuring Wavelength scan measurement (ABS
conditions measurement)
Wavelength range : 200 to 950 nm
Scan speed : 200 nm/min
Data interval : High resolution
Response : Standard
Use of data measured after baseline correction.
Excluding influence by noise, water vapor and
absorption due to quartz
Calculation method 750 points of data, excluding the top data, are
grouped into 150 blocks (1 block corresponding
to 5 nm), and the maximum value (a(i)) and
minimum value (b(i)) in “i” block are determined.
Then, flatness (c(i)) for 5 nm is calculated by the
following formula.
Flatness (c(i)) for 5 nm = (a(i) - b(i))/2 + b(i)
Flatness (c(i)) for 5 nm in all data (150 blocks) is
calculated and the maximum value (A) and
minimum value (B) of c(i) are determined.
Then, baseline flatness is calculated by the
formula given below.
Baseline flatness = ±(A - B)/2
Judgment standard Within ±0.010 Abs

5 - 14
 5.1.4

Fig. 5-18 Example Result of Baseline Flatness Check

5 - 15
5.1 Check with Instrument Main Unit Alone

5.1.5 Baseline Stability

(1) Display the screen for each item of instrument performance

check (Fig. 5-19). For display method, follow the
procedure starting from section 5.1.. Baseline stability is
required to be measured 2 hours after power-on and under
an ambient temperature change within 5 °C. Using the
[▲]/[▼] key, move the cursor to Baseline Stability, and then
select [1] Yes. Baseline stability will be checked.
If check is not desired actually, select [2] No.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
□1 Yes □
2 No

Fig. 5-19 Screen for Each Item of Instrument

Performance Check

(2) The guidance for informing the parameter setting process

(Fig. 5-20) appears, and parameters will be set and
baseline stability will be checked.

Setting PARAM....

Fig. 5-20 Guidance for Parameter Setting Process

(3) After the check of baseline stability, the screen after this
check is displayed as shown in Fig. 5-21. Measurement
result is indicated on the baseline stability line followed by
OK when the judgment standard is met or NG if not met. If
NG is indicated, check the baseline stability again in the
correct procedure. If NG recurs, refer to section 6.8.

5 - 16
 5.1.5

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability 0.0002 ABS OK
Hardware
Print Report
Select function. : _
1 Print
□

Fig. 5-21 Screen after Baseline Stability Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-7 Measuring Conditions and Judgment

Standard for Baseline Stability

Item Baseline Stability

Measuring Time scan measurement (ABS measurement)
conditions Wavelength range : 260 nm
Measurement time : 3600 s
Response : Standard
Room temperature 20 to 25 °C, at 2 hours after
power-on, temperature fluctuation within 5 °C,
noise excluded.
Use of data measured after execution of auto
zero.
Calculation method Only one smoothing is applied to the obtained
data. In the result, the maximum value (A) and
minimum value (B) of absorbance are
determined. Then, baseline stability is
calculated by the following formula.
Baseline stability (Abs/h) = A - B
Judgment standard Within 0.0007 Abs/h

5 - 17
5.1 Check with Instrument Main Unit Alone

Fig. 5-22 Example Result of Baseline Stability Check

5 - 18
 5.1.6

5.1.6 Hardware Check

(1) Display the screen for each item of instrument performance

check (Fig. 5-23). For display method, follow the
procedure starting from section 5.1.. Using the [▲]/[▼]
key, move the cursor to Hardware, and then select [1] Yes.
The hardware will be checked. If check is not desired
actually, select [2] No.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
1 Yes □
□ 2 No

Fig. 5-23 Screen for Each Item of Instrument

Performance Check

(2) After the check of RAM, ROM, lamp, wavelength driver,

wavelength and lamp usage, the hardware check screen
(Fig. 5-24) is displayed. The check result of each item is
indicated here. For printing the measurement result, press
the <1> key (Print). After check, return to the performance
check item selecting screen by pressing the [RETURN] key.

Performance CELL A
Hardware
RAM OK
ROM OK
Lamp OK
Wavelength Driver OK
WL Check OK
Lamp Usage(%) 1

Select function. : _
1 Print
□

Fig. 5-24 Hardware Check Screen

5 - 19
5.1 Check with Instrument Main Unit Alone

Table 5-8 Judgment Standard for Hardware

Item Hardware
Check items RAM : RAM is checked.
ROM : ROM is checked.
Lamp : It is checked whether the lamp
is lit or not.
Wavelength
Driver : Wavelength driver is checked.
Wavelength : Wavelength calibration at
484.6 nm is executed on the
initialization screen and it is
checked whether a peak is
detected or not.
Judgment standard OK in all check items

5 - 20
 5.1.7

5.1.7 Report Printing

(1) Display the screen for each item of instrument performance

check (Fig. 5-25). For display method, follow the
procedure starting from section 5.1.. By the Print Report
function, each item subjected to performance check can be
printed in the format of a report. For execution of this
function, select [1] Yes.

Performance CELL A
WL Accuracy (484.6 nm)
WL Accuracy (229.0 nm)
WL Accuracy (822.8 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware
Print Report
Execute(Manual)? :：＿
1 Yes □
□ 2 No

Fig. 5-25 Screen for Each Item of Instrument

Performance Check

(2) A report is printed as shown below.

Report: 2010/04/10 15:12

Spectrophotometer
Model : U-5100 Spectrophotometer
Serial No. : 1234-567
ROM VER. : 3J25300
Option :

Performance Check
WL Accuracy (484.6 nm) : -0.4 nm Tolerance ±1.0 nm OK Items subjected to
performance check
are printed in this
area.

Fig. 5-26 Example of Report Printing

5 - 21
5.1 Check with Instrument Main Unit Alone

5.1.8 Auto Check

(1) Display the screen for automatically checking the

instrument performance (Fig. 5-27). For display method,
follow the procedure starting from section 5.1.. Make sure
nothing is contained in the sample compartment and select
[1] Yes for execution of automatic performance check.

Performance CELL A
(Auto)
WL Accuracy (484.6 nm)
WL Repeatability
Noise Level (RMS)
Baseline Flatness
Baseline Stability
Hardware

Execute it? ：＿
1 Yes □
□ 2 No

Fig. 5-27 Screen for Automatic Check of

Instrument Performance

(2) Then, the guidance for confirmation of automatic printout

(Fig. 5-28) is displayed. For automatic printout, confirm
that a printer is connected, power supply is turned on and
paper has been prepared. Then, select the <1> key (Yes).
For avoiding automatic printout, select the <2> key (No).
When automatic printout is selected, it will start after
measurement of each item. However, if printout cannot
start due to a trouble on the printer or any other cause, an
error message “Cannot print out.” will be issued.
When you select [1] Abort Meas., measurement for the
current automatic performance check is aborted. Or when
you select [2] Continue Meas., printout is avoided and the
next check starts.

Print Auto? : ＿
1 Yes □
□ 2 No

Fig. 5-28 Guidance for Confirmation of

Automatic Printout

Cannot print out.：＿

1 Abort Meas. □
□ 2 Continue Meas.

Fig. 5-29 Error Message Issued when Printout is

Impossible

5 - 22
 5.1.8

(3) After start of measurement, you must wait for about 80

minutes till the end of all measurements. Upon completion
of measurement, the screen after completion of automatic
performance check (Fig. 5-30) is displayed. OK appears
when the result of each check meets the judgment standard
or NG appears if it does not meet the standard. If NG is
indicated, take a corrective measure, referring to section
5.1.1 to 5.1.6.

Performance CELL A
(Auto)
WL Accuracy (484.6 nm) -0.2 nm OK
WL Repeatability ±0.10 nm OK
Noise Level (RMS) 0.0001 ABS OK
Baseline Flatness ±0.000 ABS OK
Baseline Stability 0.0002 ABS OK
Hardware OK

Execute it? ：＿
1 Yes □
□ 2 No

Fig. 5-30 Screen after Automatic Performance Check

5 - 23
5.2 Check with Pen Type Low-Pressure Mercury Lamp Available at Option

5.2 Check with Pen Type Low-Pressure Mercury Lamp Available at Option

For performance check with a pen type low-pressure mercury

lamp, a separately available option is required. For mounting
method of this lamp, refer to the instruction manual for the
separately available option.

CAUTION
If you gaze directly at the lit Xe flash lamp, your eyes may
be damaged. This lamp emits intense ultraviolet rays.
Therefore, you must not gaze directly at the lamp.
For gazing at the lamp, put on UV-cut glasses.

(1) After startup of the instrument, press the [MAIN MENU] key
to display the Main Menu screen (Fig. 5-31). Confirm that
nothing is contained in the sample compartment and close
the lid. For selection of performance check, press the <7>
key (Performance) and then the [ENTER] key or press the
[▲]/[▼] key to select Performance followed by the [ENTER]
key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 5-31 Main Menu Screen

(2) The instrument performance check screen (Fig. 5-32)

appears. Press the <3> key (Performance Check
(Option)) and then the [ENTER] key.

5 - 24
 5.2

Performance CELL A
1 Performance Check(Auto)
2 Performance Check(Manual)
3 Performance Check(Option)

Select function. : _

Fig. 5-32 Instrument Performance Check Screen

(3) Mount the pen type low-pressure mercury lamp available at

option to the main unit. For mounting method of this lamp,
refer to the instruction manual for this option.
In succession, refer to section 5.2.1 to 5.2.3.

5 - 25
5.2 Check with Pen Type Low-Pressure Mercury Lamp Available at Option

5.2.1 Wavelength Accuracy (with Hg Lamp)

(1) Display the Performance Check (Option) screen (Fig. 5-33).

For display method, follow the procedure starting from
section 5.2.. Using the [▲]/[▼] key, move the cursor to
WL Accuracy Hg (253.7 nm), (435.8 nm) or (546.1 nm) to
be checked, and then select [1] Yes. The wavelength
accuracy at the selected wavelength with the Hg lamp will
be checked. If check is not desired actually, select [2] No.

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm)
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg
Resolution Hg (546.1 nm)
Hardware
Print Report

(Set Hg Lamp)Execute it? ：＿

□1 Yes □
2 No

Fig. 5-33 Performance Check (Option) screen

(2) The guidance for informing the parameter setting process

(Fig. 5-34) appears, and parameters will be set and
wavelength accuracy check will be carried out.

Setting PARAM....

Fig. 5-34 Guidance for Parameter Setting Process

(3) After the check of wavelength accuracy, the screen after

this check is displayed as shown in Fig. 5-35.
Measurement result is indicated on the wavelength
accuracy line; a peak difference at each wavelength of
bright line and OK when the judgment standard is met or
NG if not met. If NG is indicated, carry out wavelength
calibration by the method instructed in 5.3.1, and then
check the wavelength accuracy again in the correct
procedure. If NG recurs, refer to section 6.8.

5 - 26
 5.2.1

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm) -0.4 nm OK
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg
Resolution Hg (546.1 nm)
Hardware
Print Report

Select function. : _
1 Print
□

Fig. 5-35 Screen after Wavelength Accuracy Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key. For checking
the wavelength accuracy at other wavelength, repeat the
procedure from step (1).

Table 5-9 Measuring Conditions and Judgment

Standard for Wavelength Accuracy Hg

Item Wavelength Accuracy (at 253.7 nm)(435.8

nm)(546.1 nm)
Measuring Emission spectrum of Hg lamp bright line (with
conditions detector on the monitor side)
Wavelength of bright line from Hg lamp
253.7 nm 435.8 nm 546.1 nm
Wavelength 247.7 to 429.8 to 540.1 to
range 259.7 nm 441.8 nm 552.1 nm

Scan speed : 40 nm/min

Data interval : High resolution
Response : Standard
Spectrum to be measured
Calculation method A difference between the peak wavelength of
measured spectrum and the bright line is
determined. Then, the wavelength accuracy is
calculated by each formula in the following
table.
Wavelength of bright line from Hg lamp
253.7 nm 435.8 nm 546.1 nm
Wavelength (Obtained (Obtained (Obtained
accuracy peak peak peak
wavelength) wavelength) wavelength)
- 253.7 - 435.8 - 546.1

Judgment standard Within ±2.0 nm

5 - 27
5.2 Check with Pen Type Low-Pressure Mercury Lamp Available at Option

5.2.2 Wavelength Repeatability (with Hg Lamp)

(1) Display the Performance Check (Option) screen (Fig. 5-36).

For display method, follow the procedure starting from
section 5.2.. Using the [▲]/[▼] key, move the cursor to
WL Repeatability Hg, and then select [1] Yes. Wavelength
repeatability with the Hg lamp will be checked. If check is
not desired actually, select [2] No.

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm)
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg
Resolution Hg (546.1 nm)
Hardware
Print Report

(Set Hg Lamp)Execute it? ：＿

□1 Yes □
2 No

Fig. 5-36 Performance Check (Option) Screen

(2) The guidance for informing the parameter setting process

(Fig. 5-37) appears, and parameters will be set and
wavelength repeatability Hg will be checked.

Setting PARAM....

Fig. 5-37 Guidance for Parameter Setting Process

(3) After the check of wavelength repeatability with the Hg lamp,

the screen after this check is displayed as shown in Fig. 5-
38. Measurement result is indicated on the Hg wavelength
repeatability line followed by OK when the judgment
standard is met or NG if not met. If NG is indicated, check
the wavelength repeatability again in the correct procedure.
If NG recurs, refer to section 6.8.

5 - 28
 5.2.2

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm)
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg ±0.05 nm OK
Resolution Hg (546.1 nm)
Hardware
Print Report

Select function. : _
1 Print
□

Fig. 5-38 Screen after Wavelength Repeatability Check

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-10 Measuring Conditions and Judgment Standard

for Wavelength Repeatability with Hg Lamp

Item Wavelength Repeatability with Hg Lamp

Measuring conditions Emission spectrum of Hg lamp bright line
(with detector on the monitor side)
Wavelength range : 540.1 to 552.1 nm
Scan speed : 40 nm/min
Data interval : High resolution
Response : Standard
Spectrum to be measured 3 times.
2nd measurement after shifting to 1100 nm.
3rd measurement after shifting to 190 nm.
Calculation method A difference between maximum and minimum
peak wavelength values in 3 measurements
is determined. Then, wavelength
repeatability is calculated according to the
following formula.
Wavelength repeatability =
±(difference between maximum and minimum
peak wavelength values)/2
Judgment standard Within ±0.5 nm

5 - 29
5.2 Check with Pen Type Low-Pressure Mercury Lamp Available at Option

5.2.3 Resolution

(1) Display the Performance Check (Option) screen (Fig. 5-39).

For display method, follow the procedure starting from
section 5.2.. Using the [▲]/[▼] key, move the cursor to
Resolution Hg (546.1nm), and then select [1] Yes.
Resolution with the Hg lamp (546.1 nm) will be checked. If
check is not desired actually, select [2] No.

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm)
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg
Resolution Hg (546.1 nm)
Hardware
Print Report

(Set Hg Lamp)Execute it? ：＿

□1 Yes □
2 No

Fig. 5-39 Performance Check (Option) Screen

(2) The guidance for informing the parameter setting process

(Fig. 5-40) appears, and parameters will be set and
resolution with the Hg lamp (546.1 nm) will be checked.

Setting PARAM....

Fig. 5-40 Guidance for Parameter Setting Process

(3) After the check of resolution with the Hg lamp (546.1 nm),
the screen after this check is displayed as shown in Fig.
5-41. Measurement result is indicated on the Resolution
Hg (546.1nm) line followed by OK when the judgment
standard is met or NG if not met. If NG is indicated, check
the resolution again in the correct procedure. If NG recurs,
refer to section 6.8.

5 - 30
 5.2.3

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm)
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg
Resolution Hg (546.1 nm) 4.75 nm OK
Hardware
Print Report

Select function. : _
1 Print
□

Fig. 5-41 Screen after Resolution Check with

Hg Lamp (546.1 nm)

(4) For printing the measurement result, press the <1> key
(Print). The performance check item selecting screen
returns when you press the [RETURN] key.

Table 5-11 Measuring Conditions and Judgment Standard

for Resolution with Hg Lamp (546.1 nm)

Item Resolution with Hg Lamp (546.1 nm)

Measuring Emission spectrum of Hg lamp bright line (with
conditions detector on the monitor side)
Wavelength range : 540.1 to 552.1 nm
Scan speed : 40 nm/min
Data interval : High resolution
Response : Standard
Spectrum to be measured.
Calculation method For the peak value (A) in the determined
spectrum, peak width B (half-value width) at a
height of (A + h)/2 is obtained. The B value is
taken as a resolution.
Energy

A
(A-h)/2

0 h

Wavelength

Half-value width (B)

Judgment standard Within 5 ± 0.50 nm

5 - 31
5.2 Check with Pen Type Low-Pressure Mercury Lamp Available at Option

5.2.4 Report Printing

(1) Display the Performance Check (Option) screen (Fig. 5-42).

For display method, follow the procedure starting from
section 5.2.. By the Print Report function, each item
subjected to performance check with the Hg lamp can be
printed in the format of a report. For execution of this
function, select [1] Yes.

Performance CELL A
(Option)
WL Accuracy Hg (253.7 nm)
WL Accuracy Hg (435.8 nm)
WL Accuracy Hg (546.1 nm)
WL Repeatability Hg
Resolution Hg (546.1nm
Hardware
Print Report

Execute it? ：＿
1 Yes □
□ 2 No

Fig. 5-42 Performance Check (Option) Screen

(2) A report is printed as shown below.

Report： 2010/04/10 15:12

Model ： U-5100 Spectrophotometer
Serial No. ： 1234-567
ROM VER. ： 3J25300
Option ：

Performance Check
Items subjected to
WL Accuracy Hg (253.7 nm)： -0.4 nm Tolerance ± 2.0 nm OK option performance
check are printed in
this area.

5 - 32
 5.3.1

5.3 Wavelength Calibration

Wavelength calibration is required if the instrument performance

specification is not satisfied in the wavelength accuracy check.
The method of wavelength calibration is explained here.

5.3.1 Wavelength Calibration Method

(1) After startup of the instrument, press the [MAIN MENU] key
to display the Main Menu screen (Fig. 5-43). For selection
of system setup, press the <6> key (System Setup) and
then the [ENTER] key or press the [▲]/[▼] key to select
System Setup followed by the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 5-43 Main Menu Screen

(2) The System Setup screen (Fig. 5-44) appears.

For selecting wavelength calibration, press the <1> key (WL
CALIB.) and then the [ENTER] key or press the [▲]/[▼] key
to select WL CALIB. followed by the [ENTER] key.

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. : _

Fig. 5-44 System Setup Screen

5 - 33
5.3 Wavelength Calibration

(3) The wavelength calibration executing screen (Fig. 5-45) is

displayed. Select [1] Yes for execution of wavelength
calibration. If wavelength calibration is not desired actually,
select [2] No.

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Execute it? ：＿
1 Yes □
□ 2 No

Fig. 5-45 Wavelength Calibration Executing Screen

(4) The guidance for informing the wavelength calibrating

process (Fig. 5-46) is displayed and wavelength calibration
starts. After the calibration, you can return to the Main
Menu window by pressing the [RETURN] key.

WL CALIB.…

Fig. 5-46 Guidance for Wavelength Calibrating Process

(5) Check wavelength accuracy again as required.

5 - 34
 6.1

6. MAINTENANCE
This product is an instrument which requires periodic maintenance. In this section,
explanation is given centering on instrument cleaning, storage, specifications, etc. If use of
the instrument is continued without performing periodic checkup/maintenance, the instrument
may become faulty, which would lead to a serious trouble such as water leak, electric leak or
combustion.
Consumables and lifetime-limited parts should be purchased from your dealer or the nearest
maintenance service company authorized by Hitachi High-Technologies.

6.1 Lamp Usage

You can check the present time point in the life cycle of the lamp
in the following procedure.

(1) After startup of the instrument, press the [MAIN MENU] key
to display the Main Menu screen (Fig. 6-1). For selection
of system setup, press the <6> key (System Setup) and
then the [ENTER] key or press the [▲]/[▼] key to select
System Setup followed by the [ENTER] key.

Main Menu CELL A

1 Photometry
2 WL Scan
3 Time Scan
4 Method Menu
5 Data Menu
6 System Setup
7 Performance
8 Data Display

Select function. : _

Fig. 6-1 Main Menu Screen

6-1
6.1 Lamp Usage

(2) The System Setup screen (Fig. 6-2) appears. Press the
<2> key (Lamp Usage) and then the [ENTER] key or press
the [▲]/[▼] key to select Lamp Usage followed by the
[ENTER] key.

System Setup CELL A

1 WL CALIB.
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Select function. : _

Fig. 6-2 System Setup Screen

(3) The Lamp Usage screen (Fig. 6-3) appears. Lamp usage
will be indicated in percentage (%). The 100% value
stands for the end of service life as a standard. If 100% is
exceeded, performance check should be carried out
according to Section 5 in order to check if the lamp
performance is within the specified range. If out of the
specified range, contact your dealer or the nearest
maintenance service company authorized by Hitachi High-
Technologies.

System Setup CELL A

1 WL CALIB. Usage(%) 1
2 Lamp Usage
3 Date and Time
4 Printer Setup
5 6 Cell Mode
6 Auto Lamp OFF
7 Language

Fig. 6-3 Lamp Usage Screen

(4) After check, return to the Main Menu screen by pressing the
[RETURN] key.

6-2
 6.2

6.2 How to Clean the Instrument

a. Cleaning the Sample Compartment

Is a sample solution is spilled in the sample compartment,

remove the 6-cell holder by the determined method
(described on section 2.4.5.) and wipe off the sample
solution without delay.

<1> Turn off the instrument power supply.

<2> Pull out the power plug from the receptacle.
<3> Open the lid of the sample compartment.
<4> Loosen the screw at the top of the 6-cell holder and
remove the holder.
<5> If the spilled sample solution flows out through the
discharge port at the bottom of instrument via the
drain of sample compartment, the bottom area of the
instrument should be cleaned.

b. Cleaning the Exterior of Spectrophotometer

Before cleaning the exterior of the spectrophotometer, turn

off the power supply and pull out the power plug from the
receptacle. Then, clean the exterior always using a soft
cloth or a cloth wetted with water and wringed hard.
Avoid use of alcohol, benzine, paint thinner or the like
inflammable solvent for cleaning.
In case a sample or the like is spilled on the
spectrophotometer, turn off the power supply and pull out
the power plug from the receptacle. Then, wipe off the
spilled substance as soon as possible using a soft cloth or
a cloth wetted with water and wringed hard. You should
confirm that the wiped area is adequately dry.
Be particularly careful not to spill a sample or the like over
the control panel. If abnormality is found on the exterior,
contact your dealer or the maintenance service company
authorized by Hitachi High-Technologies.

6-3
6.3 Cleaning and Storing a Sample Cell

6.3 Cleaning and Storing a Sample Cell

For cleaning a sample cell, wash it by the determined method

using a detergent for laboratory glass apparatus. Then, rinse
the cell adequately with ultrapure water and air-dry it in a clean
environment followed by its storage.

6.4 Replacement of Radiation Source Lamp

This lamp is warranted for 1 year. The user must not attempt to
replace the lamp definitely. Entrust lamp replacement work to a
service engineer who has received training at Hitachi High-
Technologies.

6.5 Caution on Lithium Battery

CAUTION
The Model U-5100 ratio beam spectrophotometer uses a
lithium battery for memory backup. If the lithium battery is
handled in a wrong way, it might burst.
Never try its recharging, disassembly and throwing into a
flame. This battery must be disposed by a different
method from that of general wastes.
When replacement of the lithium battery becomes
necessary (for example, an error message “RAM Check
NG” is frequently issued), contact your dealer or the
maintenance service company authorized by Hitachi High-
Technologies.
Entrust this replacement work to a service engineer who
has received training at Hitachi High-Technologies (you are
charged for this work after the warranty period of the
instrument).

6-4
 6.6

6.6 Replacement of Power Fuse

WARNING
Before replacement, make sure that the power cord is not
connected.

If the power fuse is blown due to any cause, it should be

replaced with a new one in the following procedure. If the new
fuse is also blown, the instrument may be faulty. Contact your
dealer or the maintenance service company authorized by
Hitachi High-Technologies.

(1) Unplug the power cord from the connector of the

spectrophotometer. (See <1> in Fig. 6-4.)

<1>

Fig. 6-4 Unplugging the Power Cord

(2) The fuse holder is located above the power connector on

the rear side of spectrophotometer. Push the fuse holder
stopper from either side using a blade-edge screwdriver,
and the holder will come out slightly. (See <2> in Fig.
6-5.) In succession, push the other stopper in the same
manner as above, and the entire holder will advance slightly
toward you. Then, grasp and pull out the holder.

6-5
6.6 Replacement of Power Fuse

<2>

Fuse holder Power connector

Fig. 6-5 Ejection of Fuse Holder

(3) Remove the blown fuse and replace it with a new one.
(See <3> in Fig. 6-6.) Use a fuse having a proper capacity
(time lag fuse (2 A), P/N J821391). It is recommended to
prepare a spare fuse.

<3>

Fig. 6-6 Removal of Fuse Holder

6-6
 6.7

6.7 Storage of Instrument

a. After Measurement

(1) Turn off the power switch and unplug the power cord
from the receptacle.

(2) Cover the instrument with a clean cloth or the like.

NOTICE: When an organic solvent or toxic gas sample is

placed in the sample compartment, it shall be
taken out of the compartment and prevented
from being touched.

b. Before Leaving the Instrument Unused for a Long Time

(1) Store at the condition without dew condensation.

0°to 40°C and 15 to 80% humidity
(less than 70% humidity at more than 30°C) are
required.
In addition, the instrument must not receive strong
vibrations.

(2) Cover the instrument with a clean cloth or the like.

(3) Prevent acidic, alkali or any other toxic gas from

flowing into the instrument.

(4) Avoid a place where a strong magnetism may be

generated.

(5) Avoid a dusty environment.

(6) Avoid exposure to direct sunlight.

(7) The data saved in the instrument may be erased due

to complete discharge or deterioration of the lithium
battery. Therefore, you should back up the important
measurement data or conditions saved in the
instrument by printing or the like means.

6-7
6.8 If You Encounter a Trouble

6.8 If You Encounter a Trouble

If an error message appears, take a corrective measure,

referring to “a. Error Messages.” If there is an abnormality on
the instrument, take a remedial measure, referring to “b.
Troubleshooting.” If the instrument does not work normally
despite the troubleshooting, contact year dealer or the
maintenance service company authorized by Hitachi High-
Technologies.

WARNING
Electric Shock due to Contact with Instrument Interior
The instrument incorporates electric components which
might cause an electric shock if touched directly with hand.
Checkup of the instrument interior shall be entrusted to a
service engineer.

a. Error Messages

Error Message Cause Remedy

Peak not found. Peak is not found in Peaks may be below
peak detection. the threshold.
Lower the threshold
and retry peak
detection.
Printer not ready. The printer is not ready Set the printer in the
for printing. printable status.
Set the On the measurement Adjust input so that
End WL - parameter setup screen, the measuring
Start WL >= 10 nm. the measuring wavelength range is
wavelength range is less 10 nm or wider.
than 10 nm.
Set the In measurement Set the ordinate so
Upper Scale > parameters, the upper that the upper limit is
Lower Scale. limit of ordinate is equal higher than the lower
to or lower than the limit.
lower limit.
Set X-Max > X-Min. On the scale change Set the X axis so that
screen, the maximum the maximum point is
point on X axis is equal larger than the
to or smaller than the minimum point.
minimum point.
Set Y-Max > Y-Min. On the scale change Set the Y axis so that
screen, the maximum the maximum point is
point on Y axis is equal larger than the
to or smaller than the minimum point.
minimum point.

6-8
 6.8

(cont’d)
Error Message Cause Remedy
Set X-Max - On the scale change Set the X axis so that
X-Min >= 10. screen for time scan the difference between
measurement, the Max and Min is 10 s or
difference between X- more.
Max and X-Min is set to
be less than 10 s.
Set X-Max - On the scale change Set the X axis so that
X-Min >= 1. screen for wavelength the difference between
scan measurement, the Max and Min is 1 nm or
difference between X- more.
Max and X-Min is set to
be less than 1 nm.
The same CONC In calibration curve data, Different values should
value cannot be the same concentration be set.
set. value of standard
sample is set doubly.
Set a NUM of In calibration curve Set 2 or more standards
STD >=2. parameters, the number or set Thru Zero in ON
of standards is set to 1 status.
(provided Thru Zero is
OFF).
Set Upper LMT. In calibration curve Set the concentration
> Lower LMT. parameters, the upper scale so that the upper
limit of concentration is limit is higher than the
equal to or lower than lower limit.
the lower limit.
Already exists. It was attempted to save When file overwriting is
Overwrite it?: a Method file or data file not desired, change to
in an already used file an unused file name.
name.
Data will be Although data exists, When there is the
deleted. [MAIN MENU], necessary data which
Continue? [RETURN] and [START] has not yet been saved,
keys were pressed. save the relevant file
and then start up
measurement.
No. of Meas. In sample measurement Terminate
reached 150. in the photometry mode, measurement by
the maximum number of [STOP] key and save
measurements (150) the current file.
has been reached. Then, carry out the
subsequent
measurements as a
new series.
No. of files is full. The number of saved Delete unnecessary
files has reached the files, and then retry
limit (50 Method files or saving.
30 data files).
File name is not In saving, any character Save in characters
correct. (/, :, *) unallowable for a usable for a file name.
file name is used.
Set the No. of When data mode of Reduce the number of
WL <=3. photometry is wavelengths to 3 or
concentration, 4 or more less. Or change the
wavelengths have been data mode to ABS
set. or %T.

6-9
6.8 If You Encounter a Trouble

(cont’d)
Error Message Cause Remedy
Set WL1>WL2. When data mode of WL1 must be set at a
photometry is value larger than
concentration (2 WL2.
wavelengths), wavelength
value WL1 is smaller than
WL2.
Set When data mode of Set wavelength
WL1>WL2>WL3. photometry is values in the order of
concentration (3 WL1 > WL2 > WL3.
wavelengths), wavelength
values are not set in the
following order; WL1 >
WL2 > WL3.
Cannot print out. This error occurs if Retry execution after
measurement is started setting the printer in
with automatic printout the printable status.
turned ON for automatic
performance check and
then printout processing is
executed with the printer
set in printing-inhibited
status during
measurement.
Cannot load the Abnormality is found in Retry loading. If no
file. (Sum Error) loading of a Method file or improvement can be
data file. seen, contact the
maintenance service
company authorized
by Hitachi High-
Technologies.
No. of STD >=2. This error occurs if you Under the condition at
Cannot delete. attempt to set the left, the remaining
remaining number of number of standards
standards to 1 by data cannot be set to 1.
deletion in STD editing
under the condition of Thru
Zero:OFF.
Effective STD Calibration curve cannot Change to an
data is be generated due to an appropriate standard
insufficient. insufficient number of data solution and carry out
points, because there are measurement again.
the same absorbance
values in standard sample
measurement.
Set WL2 except For photometry in the DNA Specify different
CORR.WL. measurement mode, values for wavelength
wavelength 2 and 2 and correction
correction wavelength are wavelength.
set at the same value.
Set WL4 except For photometry in the DNA Specify different
CORR.WL. measurement mode, values for wavelength
wavelength 4 and 4 and correction
correction wavelength are wavelength.
set at the same value.

6 - 10
 6.8

(cont’d)
Error Message Cause Remedy
Xe Lamp Error Abnormality is found Turn on the lamp again.
when turning on the If no improvement can be
lamp. seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
Error occurred Wavelength Retry wavelength
when WL was initialization is initialization. If no
initialized abnormal. improvement can be
seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
WL Motor Error Wavelength Retry wavelength
initialization is initialization. If no
abnormal. improvement can be
seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
WL CALIB. Error Abnormality is found in Retry wavelength
wavelength calibration calibration. If no
with the bright line of improvement can be
lamp. seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
WL Correction Abnormality is found in Retry wavelength
Error wavelength correction. correction. If no
improvement can be
seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
Data Save Error RAM does not work Retry data saving. If no
normally. improvement can be
seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
6 Cell Initialize Initialization of 6-cell Mount this holder
Error holder position is properly and retry
abnormal. initialization. Or if any
holder other than the 6-
cell holder is used, set 6-
cell condition, referring to
section 2.5.4..
6 Cell Move Error Abnormality is found in Check the sample
detection of cell compartment to see if
position. there is anything that
hinders rotation of 6 cells.

6 - 11
6.8 If You Encounter a Trouble

(cont’d)
Error Message Cause Remedy
WL Motor Error Irregularity is found Retry driving. If no
when driving the improvement can be
wavelength motor. seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.
Capacitor Error Irregularity is found Retry setting. If no
when setting the improvement can be
capacitor. seen, contact the
maintenance service
company authorized by
Hitachi High-
Technologies.

b. Troubleshooting

Symptom Cause Remedy

No indication <1> The power cord <1> Plug in the power
appears even if the is unplugged. cord.
power switch is <2> The fuse is <2> Replace the fuse
turned on. blown. with a new one.
<3> Any cause other <3> If the same
than above symptom persists
after tuning off the
power switch and
turning it on, contact
the maintenance
service company
authorized by
Hitachi High-
Technologies.
Significant <1> The sample cell <1> Remove
variations in or the contaminants or
measured values transparent water drops.
plate is <2> Close the lid of
contaminated or sample
clouded with compartment.
water drops.
<3> If the same
<2> The lid of symptom persists
sample after tuning off the
compartment is power switch and
not closed. turning it on, contact
<3> Any cause other the maintenance
than above service company
authorized by
Hitachi High-
Technologies.

6 - 12
 6.8

(cont’d)
Symptom Cause Remedy
NG appears for RAM If the same symptom
RAM check on the malfunctioned. persists after tuning off the
initialization screen. power switch and turning it
on, contact the
maintenance service
company authorized by
Hitachi High-Technologies.
NG appears for ROM If the same symptom
ROM check on the malfunctioned. persists after tuning off the
initialization screen. power switch and turning it
on, contact the
maintenance service
company authorized by
Hitachi High-Technologies.
NG appears for The lamp does not If the same symptom
lamp check on the light. persists after tuning off the
initialization screen. power switch and turning it
on, contact the
maintenance service
company authorized by
Hitachi High-Technologies.
NG appears for The wavelength If the same symptom
wavelength drive mechanism is persists after tuning off the
initialization on the faulty. power switch and turning it
initialization screen. on, contact the
maintenance service
company authorized by
Hitachi High-Technologies.
NG appears for Peak wavelength Confirm that nothing is
wavelength check cannot be found. contained in the sample
on the initialization compartment. If the same
screen. symptom persists after
tuning off the power switch
and turning it on, contact
the maintenance service
company authorized by
Hitachi High-Technologies.
NG appears for The result of Carry out wavelength
wavelength wavelength calibration, referring to
accuracy in accuracy check “5.3.1 Wavelength
performance check. does not satisfy the Calibration Method (section
specification. 5.3.1.)” and check the
performance again. If no
improvement can be seen,
contact the maintenance
service company
authorized by Hitachi High-
Technologies.
NG appears for The result of Check the performance
wavelength wavelength again. If no improvement
repeatability in repeatability check can be seen, contact the
performance check. does not satisfy the maintenance service
specification. company authorized by
Hitachi High-Technologies.

6 - 13
6.8 If You Encounter a Trouble

(cont’d)
Symptom Cause Remedy
NG appears for The result of noise Check the performance
noise level in level check does again. If no improvement
performance check. not satisfy the can be seen, contact the
specification. maintenance service
company authorized by
Hitachi High-Technologies.
NG appears for The result of Check the performance
baseline flatness in baseline flatness again. If no improvement
performance check. check does not can be seen, contact the
satisfy the maintenance service
specification. company authorized by
Hitachi High-Technologies.
NG appears for The result of Check the performance
baseline stability in baseline stability again. If no improvement
performance check. check does not can be seen, contact the
satisfy the maintenance service
specification. company authorized by
Hitachi High-Technologies.
NG appears for The result of Check the performance
resolution in resolution check again. If no improvement
performance check. does not satisfy the can be seen, contact the
specification. maintenance service
company authorized by
Hitachi High-Technologies.

6 - 14
 6.9

6.9 Specifications of Model U-5010 Ratio Beam Spectrophotometer

Table 6-1 Specifications of Model U-5010 Ratio Beam

Spectrophotometer

Optics Seya-Namioka mount, ratio beam

Measuring wavelength range 190 to 1100 nm
Spectral bandpass 5 nm (546.1 nm)
Stray light 0.07% or less (220 nm for NaI,
340 nm for NaNO2)
Wavelength accuracy ±1 nm (at 484.6 nm)
Wavelength setting ±0.5 nm
repeatability
Measurement mode Photometry, wavelength scan, time
scan, multi-wavelength
measurement, ratio calculation (DNA
measurement)
Photometric range Abs : -3.000 to 3.000
%T : 0 to 300%T
Conc : 0.000 to 9999
Photometric accuracy (verified ±0.003 Abs (0 to 0.5 Abs)
with NIST SRM930) ±0.005 Abs (0.5 to 1.0 Abs)
Photometric repeatability (with ±0.002 Abs (0 to 1.0 Abs)
NIST SRM930)
Wavelength scan speed 40, 100, 200, 400, 800, 1200,
2400 nm/min (excluding filter
exchange time)
Baseline stability 0.0007 Abs/h (at 260 nm, room
temperature 20 to 25 °C,
temperature variation within 5 °C, 2
hours after power-on)
Baseline flatness ±0.010 Abs (within 200 to 950 nm,
excluding influence by noise, water
vapor and absorption due to quartz)
Noise level (RMS) 0.0002 Abs (at 260 nm, 0 Abs)
Radiation source Xenon (Xe) flash lamp
Detector Silicon photodiode
Display LCD monitor with back-light
illumination
Printer output Centronics-compatible
External dimensions 356 (W) × 426 (D) × 235 (H) mm
Weight 13 kg
Power requirements 100, 115, 220, 230, 240 V, 50/60 Hz
Power consumption 60 VA

6 - 15
 APPENDIX

Appendix a. Operating Principles of Model U-5100

Optics:
Figure “a” shows the optics of Model U-5100 ratio beam
spectrophotometer. For radiation source, a xenon flash lamp
is used. The white light emitted from the radiation source
passes through the filter and entrance slit and is converted
into a monochromatic beam by the Seya-Namioka mount
monochromator. This monochromator uses a Hitachi’s
original stigmatic concave diffraction grating with a grating
constant of 1/600 nm, blaze wavelength of 250 nm and
diffraction area of 20 mm by 25 mm. The monochromatic
beam is restricted into a bandpass of 5 nm with the exit slit,
reflected by the toroidal mirror (M2) and split into a monitor-
side (reference) beam and a sample beam. The monitor-side
beam is incident on the monitor-side detector mounted in the
monochromator. The sample beam passes through a
sample in the sample compartment and impinges upon the
sample-side detector.
The sample and reference beams incident on the respective
detectors are converted into electric signals. The Model U-
5100 detects the reference beam in order to compensate for
variations in light quantity with time. This optics is called
“ratio beam.” The ratio beam spectroscopy ensures a highly
stable photometric values unavailable with a single beam
spectrophotometer.
The beam at the center of a set 10 mm rectangular cell is 9
mm high and 3 mm wide.

APPENDIX - 1
 Xe flash lamp
Concave diffraction
grating
Mirror
Entrance slit Filter (F) (M1)

Monitor-side
detector

Exit slit
6-cell holder

Condenser
lens

Half mirror
(HM) Sample Sample-side
detector

Toroidal mirror (M2)

Monochromator Sample compartment

Fig. a-1 Optics of Model U-5100 Ratio Beam

Spectrophotometer

APPENDIX - 2
 Signal Processing and Control System:
Figure “a-2” shows the signal processing and control system
arrangement. This system is controllable with the operation
panel. After opto-electric conversion by a pair of detectors,
electric signals are amplified and A/D converted, and then
logarithmically converted by software to generate absorbance
data. The result of measurement is displayed on the data
display unit and can be printed with the printer.

Spectrophotometric
block Half mirror
Monitor-side detector
Photometric
section

Radiation source Sample

+
Monochromator Sample-side
detector

Signal and data processing block

A/D
I/O interface converter

Operation Data display Parallel

panel interface

External output block

Printer

Fig. a-2 Signal Processing and Control System

APPENDIX - 3
Appendix b. Absorption Spectrophotometry

This spectrophotometer is designed for absorption analyses of

liquid, solid and gaseous samples in the ultraviolet to visible
spectral region. Assume that a monochromatic beam with
intensity “Io” travels through a single-component sample liquid
phase having concentration “c” and path length “l,” which results
in the monochromatic radiation intensity decreasing to “It”
Then, the relation expressed by (Equation 1) will hold.
Where, ε is a constant known as absorption coefficient
(absorptivity), which varies depending on sample. And, t
denotes transmittance. Transmittance expressed as a
percentage stands for T (percent transmittance).

It
= 10−ε⋅c⋅l = t ........................(Equation 1)
I0
100 ⋅ t = T ................................(Equation 2)
1
log =ε⋅ c ⋅ l = A ....................(Equation 3)
t

Also, the common logarithm of inverse transmittance can be

expressed by (Equation 3). This equation is called “Bouguer-
Beer (Lambert-Beer) law. “A” in (Equation 3) is called
absorbance (abbreviated as Abs). Absorbance “A” is
proportional to concentration “c.” Therefore, using this
relationship, quantitative analysis is enabled by comparing
absorbance between a concentration-known standard solution
and a concentration-unknown solution. The Model U-5100 is
capable of measuring both percent transmittance and
absorbance.
For the customer who will use a spectrophotometer for the first
time, it is recommended to read JIS K 0115:2004 “General rules
for molecular absorptiometric analysis.”

Monochromatic Transmitted
beam Io beam It

Concentration c of single-
component sample
Liquid phase length l

Fig. b-1 Bouguer-Beer (Lambert-Beer) Law

APPENDIX - 4
Appendix c. Proper Use of Spectrophotometer

(1) Solvent Selection

When selecting a solvent for sample preparation, keep the

following requirements in mind.

• No or small absorption in measuring wavelength range.

• Non-interactive with solute.
• Low volatility.

Table c-1 lists the applicable wavelength ranges of common

organic solvents. (For details, refer to the new “Analytical
Chemistry” volume of Experiment Chemistry Course issued
by The Chemical Society of Japan.)
Also, the wavelength range where organic solvents are
applicable varies with grade. It is recommended to use
solvents having a grade for spectrochemical analysis.

APPENDIX - 5
 Table c-1 Applicable Wavelength Ranges (indicated
by solid lines)

Wavelength 200 nm 300 nm 400 nm or more

Solvent
200 nm
Cyclohexane
220 nm
Ethyl alcohol
220 nm
Methyl alcohol
220 nm
Diethyl ether
220 nm
Dioxane
220 nm
n-hexane
250 nm
Chloroform
250 nm
Isopropyl alcohol
250 nm
Acetic acid
270 nm
Ethyl acetate
275 nm
Carbon tetrachloride
280 nm
Benzene
2-butane 335 nm
(methyl ethyl ketone)
340 nm
Acetone
380 nm
Carbon disulfide

APPENDIX - 6
(2) Special Samples

Note that the Bouguer-Beer law mentioned in Appendix “b”

is not applicable to the following special samples.

• Fluorescing sample
• Significantly turbid sample

In measurement of a solid sample such as glass plate, the

energy of beam radiation undergoes a loss (r) due to
reflection on the surface of solid substance. This case can
be expressed by (Equation 4). “r” varies depending on the
reflectance of substance.

I t I 0 = 10 −ε ⋅c⋅l − r .........(Equation 4)

APPENDIX - 7