Model: 53F Document Type: User Manual Customer P.O.

:
Effective Date: 03/05/07 Revision Date: 09/28/07 Revision No.: 1.1

Operation Manual:
UV PhotoX Fluorescence System
Model 53F

Biotechnology Pulp & Paper Water Quality Petrochemical Pharma Chemical

Customer: Custom Sensors & Technology

531 Axminister Drive
Fenton, MO 63026
P: 636.305.0666 F: 636.305.1096

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Notice of Confidentiality

The information contained within this document is confidential and proprietary to Custom Sensors &
Technology (CST) and may be covered under existing US or CST patents pending. This information shall
not be reproduced or further disclosed, in whole or in part, to anyone other than employees of the
company purchasing the product without prior written consent from Custom Sensors & Technology.

References
Document Name: Operation Manual: UV Photo-X Fluorescence System Model 53F
Document Number: 53F

Disclaimer
The information in this document is subject to change without notice and should not be construed as a
commitment by Custom Sensors & Technology. CST assumes no responsibility for any errors that may
appear in this document. This manual is believed to be complete and accurate at the time of publication.
In no event shall CST be liable for incidental or consequential damages in connection with or arising from
the use of this manual.

All trademarks are the sole property of their respective owners.

Safety Guidelines

PLEASE READ THE INSTRUCTIONS CAREFULLY BEFORE WORKING WITH THIS PRODUCT!
This device has left our facility after careful testing of all the transmitters functions and safety features.
The functioning and operational safety of the product can only be ensured if the user observes the usual
safety precautions as well as the specific safety guidelines stated in these operating guidelines:
• Before connecting the device to the electrical supply, please ensure that the operating voltage
stated on the power supply corresponds to the voltage supplied to the unit.
• The functioning and operational safety of the instrument can only be maintained under the
climatic conditions specified in the specification section of this manual
• If the instrument is moved from warm surroundings, condensate may form and interfere with the
functioning of the instrument. In this event, wait until the temperature of the transmitter
equilibrates to the new temperature before putting the transmitter back into operation.
• Transmitter controls, maintenance and repair work must only be carried out by a suitable qualified
technician.
• If there is any reason to assume that the product can no longer be employed without a risk, it
must be set aside and appropriately marked to prevent further use.
• The safety of the user may be endangered if the instrument:
o is visibly damaged;
o no longer operates as specified
o has been damaged in transport
• If you are in doubt, the product should be sent back to the factory
• The operator of this product must ensure that the following laws and guidelines are observed
when using this product around dangerous substances:
o EEC directives
o National Fire Protection Association
o Safety data-sheets of the chemical manufacturer

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PhotoX Fluorometer
Installation and Operating Instructions
Table of Contents
Section 1: Unpacking ............................................................................................................ 5
Section 2: Hardware Installation...................................................................................... 7
2.1 Physical ................................................................................................................ 7
2.2 Optical................................................................................................................... 8
2.3 Electrical ............................................................................................................ 10
Section 3: Introduction to Fluorescence Monitoring ............................................... 12
3.1 Theory of Operation ...................................................................................... 12
3.2 Filter Based Fluorimetry .............................................................................. 12
Section 4: Glossary, Hardware Description, and Modes of Operation............. 14
4.1 Definition of Terms ........................................................................................ 14
4.2 Hardware Description ................................................................................... 15
4.3 Modes of Operation ....................................................................................... 16
4.3.1 Mode 1 Initialization.................................................................................. 16
4.3.2 Mode 2 Initialization.................................................................................. 16
Section 5: Instrument Controls....................................................................................... 18
5.1 Active or Operational Controls and Indicators.................................... 19
5.2 Factory and Troubleshooting Controls and Indicators..................... 20
5.3 Inactive Controls and Indicators .............................................................. 20
Section 6: Fluorometer Operations and Diagnostics .............................................. 21
6.1 Quick Start Operations Guide.................................................................... 21
6.2 Detailed Operations Guide.......................................................................... 25
6.2.1 First Power-Up Following Installation..................................................... 25
6.2.2 Mode 1 or Calibrated Operation ............................................................... 29
6.2.3 Mode 2 or Fixed Range Operation ........................................................... 32
6.2.4 Tips for Easy Operation ............................................................................... 34
6.3 Diagnostics and Troubleshooting ............................................................. 35
6.3.1 Diagnostic Indicators .................................................................................... 35
6.3.2 Basic Troubleshooting .................................................................................. 36
Section 7: System Specifications ................................................................................... 39
7.1 Optical Configuration .................................................................................... 39
7.2 Transmitter Specifications .......................................................................... 40
7.3 Light Source Specifications ........................................................................ 41
7.3.1 CST UVF-100 Xenon Flashlamp ................................................................ 41
7.4 Fluorescence Accessory Specifications .................................................. 42
7.4.1 Integrated Front Surface Probe Specifications ................................... 42
7.4.2 Stand-Alone Front Surface Probe Specifications ............................... 42
7.4.3 90º Fluorescence Flow Cell Specifications............................................ 43

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7.5 Application Specific Information, Notes, and Specifications ......... 44

7.5.1 Span Check Remote Operation Specifications.................................... 44
7.5.2 Enclosure Specifications .............................................................................. 44
7.5.3 Purge Kit Specifications ............................................................................... 44
7.5.4 Special Operational Procedures................................................................ 44
7.5.5 Additional Comments.................................................................................... 44
Appendix 1: Application Engineering Assistance...................................................... 45
A1.1 Beginning Work ................................................................................................ 45
A1.2 Mode 1 (Calibrated Analysis)..................................................................... 46
A1.3 Mode 2 (Fixed Range Analysis) ................................................................ 46
A1.4 Useful Equations ............................................................................................. 47
Table 1: Data organization example................................................................. 48
Appendix 2: Advanced Troubleshooting ...................................................................... 49
A2.1 Emergency Troubleshooting Procedure ................................................. 49
Appendix 3: Analyzer System Engineering Drawings ............................................ 50
Appendix 4: QA/QC Documentation.............................................................................. 51

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Section 1: Unpacking
CST provides the 53F system in three ways: 1) Laboratory system mounted on a back
panel, 2) Process equipment mounted in an appropriate enclosure, and 3) OEM component
hardware. In all of these configurations it is critical to proper installation and setup that the
following parts be identified.

1) Find/identify the following components

a) 53F UV PhotoX transmitter

b) Xenon flash lamp assembly

c) Fiber optic cables (2) [only included with the stand

alone probe]

d) The fluorescence probe (2 types available)

i) Integrated probe: the probe insertion length and fiber optic cables are integrated into
one assembly

ii) Stand alone: the probe has no fiber optic cables attached

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e) The cable to connect the PhotoX transmitter to the light source

i) For panel mounted systems the lamp and transmitter are already connected.
ii) For OEM components, the cable is typically shipped separated from both the light
source and the transmitter.

f) Calibration bottle

g) Adjustment/Initialization tool

2) Determine where to install/place the system

3) Insure that required resources are available (power, purge air, etc.)

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Section 2: Hardware Installation

In addition to the physical, optical, and electrical installation requirements of the fluorometer it is
recommended that the transmitter be placed in a clean, dry area. If the area surrounding the
analyzer system exceeds our upper temperature limit of 115º F, it is recommended that the
transmitter be moved. For systems mounted in an enclosure, the enclosure may be purged with
clean, dry, oil free air (or nitrogen) to help dissipate heat within the enclosure.

Please follow the outlined sections below as a guideline to installing the 53F system for your
application or in the case of OEM components onto a suitable panel.

DO NOT supply power to the unit until specifically told to do so in the Instrument Setup
and Initialization procedure given in Section 6.

2.1 Physical

Consult Appendix 3 for mechanical drawings to assist with the installation.

1. For permanent installations, bolt the fluorometer, back panel, or enclosure (as
appropriate) in place using mounting screws. The location should be secure, rigid, and
strong enough to support the weight of the installed system.
2. For laboratory use, the system is supplied on either a back panel with rubber feet or
mounting rack. Make sure there is sufficient space on a bench/table top to
accommodate the size of the hardware supplied.
3. Run required electrical wiring for instrument power and 4-20mA output signals. Review
Section 2.3 Electrical below prior to making electrical connections.
4. Run required dry and oil free purge air for the unit. (Only necessary with systems
mounted in an enclosure).
5. Run the fiber optic cable(s) in conduit or other appropriate protective measure as
desired. Refer to Section 2.2 Optical below for information on connecting the optical
components.

NOTE: Secure the fiber optic cables so that they are not free to move around, be
accidentally stepped on, leaned against, or otherwise damaged during normal
operations. The allowed bend radius for fibers supplied with the 53F system is eight
(8) inches.

6. Install probe in the sensing location, making sure there are no fluid leaks and the probe
has been fully tested under process conditions for temperature and pressure safety.
The sensing location could be the CST supplied calibration bottle for OEM and
laboratory based systems.

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2.2 Optical

Light Source SMA 905 Connection Point

(to the probe input)

53F Transmitter SMA 905 Connection Point

(from the probe output)

1. Remove the protective boots (typically black) from the SMA 905 connectors on the
fiber optic cables supplied for use with the stand alone probe or located on the
integrated probe.
2. Clean the fiber ends using a lint free cotton swab dipped in either spectroscopic
grade isopropyl alcohol (IPA, also referred to as 2-propanol) or methanol.
Spectroscopic grade acetone can also be used, but may de-laminate the cotton from
the swab by dissolving the binding agent.
3. Remove the red protective boots from the SMA 905 connections on the transmitter
and light source. For installations with the stand alone probe, there are also red
protective boots on the two probe SMA 905 connections.
4. Connect the probe to the transmitter and light source.
a. Integrated probe.
i. The fiber bundle is connected to the light source.
ii. The single fiber is connected to the transmitter.

NOTE: Pointing the probe at the over-head light will illuminate the fiber. The
single fiber will appear as a single bright spot, whereas the bundle will clearly
show several small bright spots. The fiber bundle also appears larger than the
single fiber. CST will usually label the fiber bundle as either “Bundle” or “Source”
to facilitate proper connection.

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SMA Protective boots

Input ( I )
(bundle, lamp
excitation)
Port
Output ( O )
(Return, emission,
transmitter)
Port

b. Stand alone probe.

i. Connect one fiber optic cable between the light source and the probe
input port. The probe connection is marked with an “I”.
ii. Connect one fiber optic cable between the transmitter and the probe
output port. The probe connection is marked with an “O”.

NOTE: Secure the fiber optic cables so that they are not free to move around, be
accidentally stepped on, leaned against, or otherwise damaged during normal
operations. The allowed bend radius for fibers supplied with the 53F system is eight (8)
inches.

NOTE: NEVER use a wrench, pliers, or other tool to tighten the SMA connections. The
connectors are designed to operate after being finger tightened. Over tightening the
SMA connections will result in damage to the connector and the fiber optic cable limiting
the light transmission capabilities of the system and typically requiring replacement of
the fiber optic cables for normal analyzer operation.

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2.3 Electrical

Consult Appendix 3 for electrical connectivity drawings to assist with the installation.

53F transmitter electrical connectivity points are illustrated.

TB1 = power and 4-20

mA current
loop output)
(removable
terminal block)
TB2 = external/local
span control
(removable terminal
block) Grounding
Terminal

Example of terminal strip connectivity on panel mounted systems.

NOTE: A jumper (green circles above) is required if an external or remote Form B

contact (normally closed) switch or relay is not installed.

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1. Verify electrical connection between the light source and the transmitters
a) The 53F transmitter controls the light source flash rate.
b) Verify that there is a cable connection between the transmitter and light source module.
2. Connect power and ground to the system. Make sure that this step does NOT power up
the 53F transmitter. It is recommended that an ON/OFF switch be employed.
a) For panel mounted systems CST provides a terminal strip for electrical connectivity,
either 120 VAC (for systems equipped with an AC to 24 VDC power supply) or 24 VDC
per the specifications of the customer.
b) For laboratory systems, CST supplies the system with an attached 3-pronged 120V AC
power cord. CST typically installs an ON/OFF switch on these systems; however, the
customer may request this option be eliminated.
3. Connect the 4-20 mA signal output line.
a) For panel mounted systems CST provides a terminal strip for electrical connectivity.
b) For OEM systems, connectivity must be made to terminals on the TB1 connector
(reference the appropriate schematic in Appendix 3)
4. Span Check Filter Control
a) For panel mounted systems, verify that there is a connection on the terminal strip to
either an external normally closed relay or that a jumper wire is connected between the
terminal points (reference Appendix 3 for the electrical connectivity schematic). Failure
to have the connection in place will prevent the instrument from initializing.
b) Insure that there is either a jumper or connections to a terminal strip or the TB2
connector (top of the transmitter) between the COM and SPAN N.C. positions. Failure
to have the connection in place will prevent the instrument from initializing.
5. Insure that both the 24V Power and the Loop Output are both properly grounded through
Terminal 3 of the TB1 connector. Improper grounding could result in erroneous
measurement results and/or measurement instability.

NOTE: DO NOT power up the system at this time. Please reference Section 6 for
instructions on powering the unit and initializing the system.

Operational Tip: CST recommends that data logging of the 4-20 mA output from the 53F
system be done with a minimum logging interval of 4 seconds. Logging on faster time scales is
not necessary since the update period for the transmitter is 4 seconds.

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Section 3: Introduction to Fluorescence Monitoring

3.1 Theory of Operation

The ability to monitor the concentration of an analyte in a process stream is critical for
accurate and reliable process control. There are many techniques used to determine the
analyte concentration of interest. One of the most sensitive sensing techniques is molecular
fluorescence. Fluorescence occurs when a molecule absorbs light energy, either ultraviolet or
visible, and rapidly emits light, at longer wavelengths. Fluorescence of this type is commonly
referred to as Stokes fluorescence. Fluorimetry characterizes the excitation and emission
properties of the molecular species. Figure 1 shows an example of the excitation and emission
spectrum from a hypothetical fluorophore.

Figure 1: Stokes fluorescence from a

fluorescent molecule.

Fluorimetry is concerned with three types of information: 1) The intensity versus

wavelength distribution, which is characteristic of the electronic properties of the molecule, 2)
The intensity of the fluorescence, which is typically correlated to the concentration of the
fluorescent molecule in the solution, and 3) The time required for fluorescence to cease (the
fluorescence lifetime). The CST 53F fluorometer is a filter-based analyzer and is only
concerned with items 1 and 2 above.

3.2 Filter Based Fluorimetry

A filter-based fluorometer is a good choice when qualitative measurements, for example
the presence or absence of the analyte of interest, are desired. Additionally, through the use of
calibration standards and environmental control, a filter based fluorometer is capable of
quantitative determination of analyte concentration. It is critical for quantitative measurements,
that the filter based analyzer is accurately correlated to standard laboratory methods and
solutions.
The 53F fluorometer uses optical filters to provide a specific range of both excitation and
emission wavelengths tuned to coincide with analyte specific molecular fluorescence. In the

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CST fluorometer the filters are located internal to the transmitter and are specific to the
application. Therefore, the CST fluorometer is a dedicated instrument for monitoring only one
specific analyte of interest. In order to monitor a new analyte of interest, the user must return
the fluorometer to CST for factory reconfiguration.
In brief, the CST fluorometer works as follows: The light source launches excitation light
through the excitation wavelength selection filter and then into a fiber optic cable. The fiber
optic cable transfers the excitation light to the CST fluorescence probe mounted in the process.
The fluorescence probe launches light into the process sample, and collects the molecular
fluorescence (emission) from the sample. The emission light is then transferred from the probe
through fiber optic cable to the analyzer. The emission light passes through an emission filter in
order to remove any residual excitation energy collected by the fluorescence probe. The
emission light then impinges a detector and the fluorescence intensity is displayed on the
analyzer.
Unlike many fluorescence units on the market, the CST fluorometer utilizes a xenon
flash lamp to provide excitation energy. This lamp allows the CST fluorometer to be easily
configured to meet any excitation wavelength requirement within the UV and visible spectral
range with the appropriate filter selection. The xenon flash lamp also has an extended lifetime
compared to other common UV sources (deuterium, mercury vapor, etc), which reduces the
cost of ownership of the analyzer.
The remainder of this manual provides the user with the necessary tools to operate the
fluorometer and meet the measurement requirements. In addition to standard operation
methods and procedures, a section detailing some application development objectives
(Appendix 1) has been provided to aid the user in defining the parameters required to realize
optimal process monitoring.

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Section 4: Glossary, Hardware Description, and Modes of Operation

4.1 Definition of Terms

Excitation Filter: The excitation filter is used to select the range of wavelengths, commonly
referred to as the pass band, used to cause or excite the molecular fluorescence. Wavelengths
not in the pass band are rejected and ideally never reach the sample. Removal of wavelengths
outside the pass band minimizes the possibility of false fluorescence readings due to detection
of light from the lamp which mimics the fluorescence signal.

Emission Filter: The emission filter is used to select the range of wavelengths, the pass band,
to be passed to the measure detector. Wavelengths not in the pass band are rejected and
ideally never reach the sample. It is critical that the excitation wavelengths never reach the
detector, since it will respond to excitation light. Excitation light impinging the measure detector
results in increased residual background levels, which reduces the dynamic range, the signal-to-
noise ratio and the signal-to-blank ratio.

Measure Detector: The light detector is most often a photomultiplier tube, though photodiodes
are increasingly being used. The light passing through the emission filter is detected by the
photomultiplier or photodiode. The light intensity, which is proportional to the analyte
concentration, is registered as a digital readout.

Span Filter: The span filter is used to check instrument operation. The span check filter used
by CST is a neutral density filter that attenuates the fluorescence signal. The degree of
attenuation is called the attenuation factor and this value is recorded in Section 7.1.

Background Fluorescence: The fluorescence signal due to the probe/analyzer optical

configuration, stray light, and fluorescence from the background material.

Sensitivity: The ability of the analyzer to detect a given level of analyte based on the molecular
fluorescence from the analyte. The actual limits of detection depend on the properties of the
analyte measured and the process conditions. Parameters such as pH, temperature, oxygen
content, and background solvent, to name but a few, may dramatically alter the fluorescence
intensity measured. Typically, detection of parts-per-million (PPM) and parts-per-billion (PPB)
analyte levels can be detected. In general, fluorescent measurements are 1,000 to 500,000
times more sensitive than absorbance based photometric measurements. Practically, sensitivity
means the minimum analyte concentration that can be measured above background
fluorescence in the process.

Specificity: The ability of the analyzer to monitor one specific analyte in a mixture of
background materials without interference from the background materials. In absorbance based
photometric measurements, interference problems are common since many materials absorb
light, making it difficult to isolate the targeted analyte in a complex mixture. However,
fluorometers are highly specific and less susceptible to interferences because fewer materials
exhibit molecular fluorescence. Furthermore, if background materials do absorb and emit light,
it is rare that they will emit the same wavelength of light as the analyte of interest.

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Signal-to-noise ratio: Signal refers to the emission collected by the fluorescence probe and
monitored by the analyzer. Noise refers to the output from the instrument’s electronics, which is
present whether or not sample is being read and any collection of errant wavelengths not
removed by the optical filters. Noise is measured by placing the fluorescence probe in air and in
complete darkness (no stray light). CST determines the signal-to-noise as a quality assurance
tool to insure that the analyzer system meets design criteria. For process monitoring, the
signal-to-noise ratio is not as important as the signal-over-background ratio.

Signal-over-background ratio: Signal refers to the emission collected from a sample with
known analyte concentration by the fluorescence probe and monitored by the analyzer.
Background refers to the process liquid containing no analyte of interest and any stray light
present in the system. The signal-over-background ratio should be calculated during the
application engineering phase of the project. Knowing this ratio will help determine when the
stray light level changes and/or the background material fluorescence properties change. Refer
to Appendix 1 for additional details.

Dynamic Range: Dynamic range refers to the range of concentrations an instrument can read,
from the minimum to the maximum detectable concentration. The minimum detectable
concentration is determined by signal-to-noise and signal-over-background ratios. The
maximum detectable concentration is determined by the compound’s chemistry and by factors
such as instrument sensitivity ranges, optical pathlength, specificity of optical filters, etc.

Linearity: Fluorescence intensity is typically directly proportional (linear) to concentration. There

are, however, many factors that affect this linear relationship. For example, variations in
temperature, pH, dissolved oxygen content, stray light, turbidity, variation in the chemical
composition of the background, etc can dramatically affect the linearity of the fluorescence
response. Practically, the linearity of the measurement is determined during the application
engineering phase of the project. CST experience indicates that fitting of the fluorescence
response to a second order polynomial provides the best concentration prediction during
process monitoring. Refer to Appendix 1 for details.

Process Background: The liquid solution used to transport or sustain the analyte of interest in
the process. This solution has all the chemical constituents found in the process except the
analyte of interest.

4.2 Hardware Description

Stand Alone Probe: CST front surface fluorescence probe, in which the fiber optic cable
connecting the probe to the transmitter is NOT connected to the probe. i.e. connecting the probe
to the transmitter requires four connections: 1) Two (2) on the probe, 2) One (1) to the light
source, and 3) One (1) to the 53F transmitter. This probe has SMA 905 connections on the
probe marked with an “I” for input (from the light source) and “O” for output (return to the 53F
transmitter).

Integrated Probe: CST front surface probe, in which the fiber optic cables are integrated into
the probe assembly, i.e. the only connections required for this probe are at the light source and

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the transmitter, not on the probe itself. Typically, the light source connection will be marked with
a label reading “Bundle” or “Source”.

Light Source: CST typically uses a Xenon flash lamp for this system, however any light source
(deuterium lamp, tungsten halogen lamp, LED, laser diode, etc.) could be used with the 53F
transmitter.

4.3 Modes of Operation

4.3.1 Mode 1 Initialization

Mode 1 initialization is a method for setting up the fluorometer for process monitoring. It
is also typically referred to as the Calibrated Mode. Mode 1 initialization is used to setup the
fluorometer when the fluorescence intensity versus analyte concentration and the background
fluorescence of the process have been characterized. Mode 1 initialization must be performed
while the fluorescence probe is immersed in the calibration (reference) sample.

In Mode 1, the fluorescence calibration is based on a process sample or standard

solution with a known analyte concentration and hence known fluorescence level. The
background material for this sample must be the process background material. The
fluorescence from the calibration (reference) sample is then used to set the midpoint of the
fluorescence scale. During Mode 1 initialization, the zero fluorescence level is set electronically
to 0 counts (display value), the midpoint range is set to 1000 counts (display reading after
initialization), and the upper limit of the range is set to 1999 counts. Section 6 details the
procedure for Mode 1 initialization.

In Mode 1, the background fluid will typically exhibit a non-zero display value. This is
why the CST fluorescence analyzer is a signal-over-background measurement. In Mode 1
operation, the 4-20 mA output response is scaled from 4 mA at 000 counts and 20 mA at 1999
counts, with the 1000 count calibration point equaling 12 mA.

NOTE: If the fluorescence of the process has not been characterized, Mode 2
initialization must be performed first followed by an application engineering study, refer
to Appendix 1 for guidance. This process is an advanced use of the 53F system, and
should only be performed in consultation with CST technical staff.

4.3.2 Mode 2 Initialization

Mode 2 initialization is a method for setting up the fluorometer for process monitoring
and/or application engineering studies. Mode 2 initialization is used to setup the fluorometer
when the fluorescence intensity versus analyte concentration and the background fluorescence
of the process are unknown. Characterization of the fluorescence response as a function of
analyte concentration can be performed onsite, refer to Appendix 1 for details, or at CST in the
Applications Development lab for a nominal charge.

Mode 2 operation electronically determines the fluorescence scale. Initialization may be

performed while the fluorescence probe is immersed in any process sample, although for

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simplicity CST recommends using the process background material without any analyte of
interest (commonly referred to as the zero or blank sample). Mode 2 initialization sets the
absolute zero fluorescence level to a display value of 000 counts and the upper fluorescence
limit to a display value of 1999. In general, the background fluorescence level will not equal 000
counts. The background level is a combination of fluid native fluorescence and the native
fluorescence of the optical materials in the probe assembly. Probe fluorescence is most
prominent when the excitation wavelength is below 280 nm. Section 6 details the procedure for
Mode 2 initialization.

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Section 5: Instrument Controls

A description of the numbered controls and indicators are listed below and broken into
three categories: 1) Active (used during normal operations), 2) Troubleshooting (only used by
CST personnel, CST certified technicians, or at the request of a CST technical staff member),
and 3) Inactive controls (these controls have no relevance or function during both normal
operations and troubleshooting).

Transmitter Front Panel

(2)

(1) (3)

(4) (5)

(6)

(7)

(8) (9) (10) (11)

Transmitter Upper Panel (Top of the Unit)

(12) (13)

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5.1 Active or Operational Controls and Indicators

During normal operations, the following controls and indicators are used for either
configuring the analyzer or monitoring operations.

Label # Description Function/Use/Operational Information

Initialization/Preset 1) Initializes analyzer operation.
1
Control Button 2) Sets the mode of operation
Indicates position of the span check filter:
Span Check LED
2 Red = In the optical path (check/verify operation)
Indicator
Green = Out of the optical path (standard operation)
Indicates the amount of energy impinging the fluorescence
emission detector

Monitoring States:
Measure Detector
4 RED = too much light intensity contact CST
Status Indicator
Solid Green – acceptable
Solid Yellow – acceptable
Flashing Green – acceptable
Flashing Yellow – acceptable
Indicates the amount of energy impinging the reference
(excitation) detector

Monitoring States:
Reference Detector
6 RED = too much light intensity contact CST
Status Indicator
Solid Green – acceptable
Solid Yellow – low light intensity contact CST
Flashing Green – low light intensity contact CST
Flashing Yellow – low light intensity contact CST
Displays the fluorescence level in counts (arbitrary units)
Format XXXX (for a range of 000 to 1999)
7 Digital Display
Values outside the format range indicates an operational
failure (contact CST)
Span Check Filter Press, hold 1-2 seconds, and then release to initiate span
12
Motion Control check filter motion, either in or out.

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5.2 Factory and Troubleshooting Controls and Indicators

These controls are used by the factory to configure and setup the analyzer. These
controls should only be used at the request of a CST technical staff member or as part of
troubleshooting procedures supplied by CST technical staff, or located in Appendix 2. NEVER
USE THESE CONTROLS DURING NORMAL OPERATIONS

Label # Description Function/Use/Operational Information

Used to adjust the zero fluorescence level to 000 counts.
8 Fine Zero Adjustment This control should NEVER be used during normal
operation.
Used to make large adjustments to the zero fluorescence
Course Zero
9 level. This control should NEVER be used during normal
Adjustment
operation.
Used to adjust the 4 mA output level to coincide with the
4 mA Level 000 fluorescence level. The 4 mA adjustment should
10
Adjustment NEVER be used to set a non-zero fluorescence level to 4
mA.
Used to adjust the 20 mA output level to coincide with the
20 mA Level 1999 fluorescence level. The 20 mA adjustment should
11
Adjustment NEVER be used to set a fluorescence level less than 1999
counts to 20 mA.
Press and hold to determine the analyzer offset from 000
counts. During normal operation the displayed value
Zero Level Check
13 should be 000 +/- 20 counts. If the value exceeds this
Switch
range, the zero levels are no long adjusted properly and
measured responses are suspect.

5.3 Inactive Controls and Indicators

These controls are inactive and have no control or operations function with regards to
the fluorescence analyzer.

Label # Description Function/Use/Operational Information

USER Mode Indicator Not in use, if active analyzer is not functioning properly,
3
LED contact CST immediately for assistance
USER Mode Span No function.
5
Adjustment

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Section 6: Fluorometer Operations and Diagnostics

This section is broken up into two parts: 1) The Quick Start Operations Guide and 2)
The Detailed Operations Guide. The Quick Start section is sufficient for all applications where
Mode 1 or calibrated operation is desired (90% of all applications). For users performing
application development work, it may be necessary to operate in both Mode 1 and Mode 2, so
detailed procedures are supplied for working in both Modes and switching between Modes.
Incorrectly switching between operational Modes can lead to significant operational difficulties
and instrument response instability.

Operation Note: The digital display and 4-20 mA output are correlated to fluorescence
intensity in counts (arbitrary units). To convert fluorescence intensity to relevant engineering
units the 4-20mA output must be scaled using an external device. The transmitter display
cannot be set to relevant engineering units, and has a fixed range of 000 to 1999 counts.

Please reference Section 2 for details regarding hardware installation (physical

mounting, optical connectivity, and electrical connectivity).

6.1 Quick Start Operations Guide

This procedure assumes that the hardware has been installed correctly and is
ready for operation. For OEM components, wiring and mounting will be required before
this procedure is applicable. For a panel mounted system this procedure is valid once
electrical connectivity is established.

DO NOT power-up the unit until directed to do so in this procedure. Powering the
unit incorrectly will result in unstable measurements and errant readings.

CST shipped the unit in Mode 1 or the calibrated mode, after completion of QA/QC
testing.

TB 2 Connector TB 1 Connector

External Span Control Grounding

Jumper (shown) Terminal

1) Insure that there is either a jumper or connections to a terminal strip on the TB2 connector
(top of the transmitter) between the COM and SPAN N.C. positions. Failure to have the
connection in place will prevent the instrument from initializing.

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2) Insure that both the 24V Power and the Loop Output are both properly grounded through
Terminal 3 of the TB1 connector. Improper grounding could result in erroneous
measurement results and/or measurement instability.
3) Connect the probe to the transmitter and light source.
a) Integrated probe.
i) The fiber bundle is connected to the light source.
ii) The single fiber is connected to the transmitter.

NOTE: Pointing the probe at the over-head light will illuminate the fiber. The single fiber
will appear as a single bright spot, whereas the bundle will clearly show several small
bright spots. The fiber bundle also appears larger than the single fiber.

b) Stand alone probe.

i) Connect one fiber optic cable between the light source and the probe. The probe
connection is marked with an “I”.
ii) Connect one fiber optic cable between the transmitter and the probe. The probe
connection is marked with an “O”.

NOTE: Secure the fiber optic cables so that they are not free to move around, be
accidentally stepped on, leaned against, or otherwise damaged during normal
operations. The allowed bend radius for fibers supplied with the 53F system is eight (8)
inches.

4) Place the probe in a dark environment, containing a non-fluorescing sample (air, water,
etc.). Examples of appropriate environments are:
a) The calibration bottle (with the probe approximately ¼ - ½” from the bottom of the bottle),
or
b) A dedicated sampling point (slip stream, side vessel, etc.), or
c) The process pipe, vessel, etc.

5) Power up the analyzer and wait a minimum of 15 minutes before proceeding with the
procedure.

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Span Filter Indicator

Measure Detector Status

Reference Detector Status

6) After the waiting period, the

following should be observed:
a) The Span Filter Indicator should be green (filter is out)
b) The Detector Status LEDs (located to the left of the display) should be:
i) Measure: Flashing Yellow or Flashing Green (typically)
ii) Reference: Solid Green
c) The display should be reading a number between 000 and 250 (typically) with a stability
of +/- 20 counts. Consult the CST QA/QC documentation for actual test results on non-
fluorescing materials (designated as Mode 1).
d) If either Detector Status LED is RED, contact CST immediately for assistance.

NOTE: The Measure Detector Status LED has four (4) allowed states for proper
fluorescence measurement. Flashing Yellow, Flashing Green, Solid Yellow and Solid
Green. If any of these four states occur, the unit should be functioning normally.

NOTE: NEVER allow the probe tip to be exposed to ambient light for longer than 1
minute. If exposure occurs, place the probe back in a dark environment for 2 hours and
suspend all work until the time has passed. Excessive light will result in RED indicator
LEDs on both the Measure and Reference Detector Status.

7) Add calibration fluid to the probe insertion region. Wait 2 minutes, the following should be
observed:
a) The Span Filter Indicator should be green (filter is out)
b) The Detector Status LEDs (located to the left of the display) should be:
i) Measure: Solid Yellow or Solid Green (typically)
ii) Reference: Solid Green
c) Assuming that the CST QA/QC calibration fluid is a relatively close match to your
calibration fluid, the display should be reading between 900 and 1100 counts (typically)
with a stability of +/- 20 counts. If you did not specify a calibration fluid, then agreement
between the factory calibration and your calibration sample is unknown. Consult the
CST QA/QC documentation for a description of the calibration solution and the actual
test results (designated as Mode 1).
d) If either Detector Status LED is RED, contact CST immediately for assistance.

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Span Filter Indicator

Initialization Button (requires

insertion of the adjustment tool)

Measure Detector Status

Reference Detector Status

8) Calibrate the analyzer to your

calibration solution.
a) Insert the Adjustment/Initialization tool into the hole marked INT PRESET and depress
the button for 3-5 seconds.
b) Wait 2 minutes, the following should be observed:
i) The Span Filter Indicator should be green (filter is out)
ii) The Detector Status LEDs (located to the left of the display) should be:
(1) Measure: Solid Yellow or Solid Green (typically)
(2) Reference: Solid Green
iii) The display should read 1000 +/- 20 counts and be stable within this allowed range.
iv) If either Detector Status LED is RED, contact CST immediately for assistance.

NOTE: If the display does not fall within the range from 980 to 1020 counts repeat the
calibration step above. Contact CST for technical assistance if three (3) attempts at
calibration are not successful.

9) You are now ready to proceed with either lab work or process monitoring.

Operation Tip 1: The 53F system is designed for continuous 24/7 operation, so leave the
system running whenever possible. Cycling of power frequently will require calibration steps on
a shorter time interval.

Operational Tip 2: The calibration bottle supplied by CST is a common Nalgene™ bottle
supplied by almost every laboratory supply company. If you are using the bottle for calibration
and system validation, purchase a set of bottles from Fisher to insure that cross contamination
between samples does not occur.

Operational Tip 3: The span check filter can be used to provide additional points during
instrument testing and verification. The filter utilized is a neutral density filter and attenuates the
fluorescence response from any sample. Consult the CST QA/QC documentation for the exact
attenuation level in your instrument.

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6.2 Detailed Operations Guide

This procedure assumes that the hardware has been installed correctly and is
ready for operation. For OEM components, wiring and mounting will be required before
these procedures are applicable.

DO NOT power-up the unit until directed to do so in the procedures outlined

below. Powering the unit incorrectly will result in unstable measurements and errant
readings.

CST shipped the unit in Mode 1, or the calibrated mode, after completion of
QA/QC testing. Whenever possible, please leave the unit in Mode 1 when powering the
unit off. Mode 1 operation is the standard or normal mode of operation.

6.2.1 First Power-Up Following Installation

TB 2 Connector TB 1 Connector

External Span Control Grounding

Jumper (shown) Terminal

1) Insure that there is either a jumper or connections to a terminal strip on the TB2 connector
(top of the transmitter) between the COM and SPAN N.C. positions. Failure to have the
connection in place will prevent the instrument from initializing.
2) Insure that both the 24V Power and the Loop Output are both properly grounded through
Terminal 3 of the TB1 connector. Improper grounding could result in erroneous
measurement results and/or measurement instability.
3) Verify the connection between the probe, transmitter, and light source.
a) Integrated probe.
i) The fiber bundle is connected to the light source.
ii) The single fiber is connected to the transmitter.

NOTE: Pointing the probe at the over-head light will illuminate the fiber. The single fiber
will appear as a single bright spot, whereas the bundle will clearly show several small
bright spots. The fiber bundle also appears larger than the single fiber.

b) Stand alone probe.

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i) Connect one fiber optic cable between the light source and the probe. The probe
connection is marked with an “I”.
ii) Connect one fiber optic cable between the transmitter and the probe. The probe
connection is marked with an “O”.

NOTE: Secure the fiber optic cables so that they are not free to move around, be
accidentally stepped on, leaned against, or otherwise damaged during normal
operations. The allowed bend radius for fibers supplied with the 53F system is eight (8)
inches.

4) Place the probe in a dark environment, containing a non-fluorescing sample (air, water,
etc.). Examples of appropriate environments are:
a) The calibration bottle (with the probe approximately ¼ - ½” from the bottom of the bottle),
or
b) A dedicated sampling point (slip stream, side vessel, etc.), or
c) The process pipe, vessel, etc.
5) Power up the analyzer and wait a minimum of 15 minutes before proceeding with the
procedure.

NOTE: CST recommends at least one (1) hour of warm-up when using the analyzer
system after an extended (>1 hour) power down period.

6) After the waiting period, the following should be observed:

a) The Span Filter Indicator should be green (filter is out)
b) The Detector Status LEDs (located to the left of the display) should be:
i) Measure: Flashing Yellow or Flashing Green (typically)
ii) Reference: Solid Green
c) The display should be reading a number between 000 and 250 (typically) with a stability
of +/- 20 counts. Consult the CST QA/QC documentation for actual test results on non-
fluorescing materials (designated as Mode 1).
d) If either Detector Status LED is RED, contact CST immediately for assistance.

NOTE: The Measure Detector Status LED has four (4) allowed states for proper
fluorescence measurement. Flashing Yellow, Flashing Green, Solid Yellow and Solid
Green. If any of these four states occur, the unit should be functioning normally.

NOTE: NEVER allow the probe tip to be exposed to ambient light for longer than 1
minute. If exposure occurs, place the probe back in a dark environment for 2 hours and
suspend all work until the time has passed. Excessive light will result in RED indicator
LEDs on both the Measure and Reference Detector Status.

7) Add calibration fluid to the probe insertion region. Wait 2 minutes, the following should be
observed:
a) The Span Filter Indicator should be green (filter is out)
b) The Detector Status LEDs (located to the left of the display) should be:
i) Measure: Solid Yellow or Solid Green (typically)
ii) Reference: Solid Green

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c) Assuming that the CST QA/QC calibration fluid is a relatively close match to your
calibration fluid, the display should be reading between 900 and 1100 counts (typically)
with a stability of +/- 20 counts. If you did not specify a calibration fluid, then agreement
between the factory calibration and your calibration sample is unknown. Consult the
CST QA/QC documentation for a description of the calibration solution and the actual
test results (designated as Mode 1).
d) If either Detector Status LED is RED, contact CST immediately for assistance.

NOTE: The Measure Detector Status LED has four (4) allowed states for proper
fluorescence measurement. Flashing Yellow, Flashing Green, Solid Yellow and Solid
Green. If any of these four states occur, the unit should be functioning normally.

NOTE: NEVER allow the probe tip to be exposed to ambient light for longer than 1
minute. If exposure occurs, place the probe back in a dark environment for 2 hours and
suspend all work until the time has passed. Excessive light will result in RED indicator
LEDs on both the Measure and Reference Detector Status.

Span Filter Indicator

Initialization Button (requires

insertion of the adjustment tool)

Measure Detector Status

Reference Detector Status

8) Calibrate the analyzer to your

calibration solution. (Mode 1 Operation)
a) Insert the Adjustment/Initialization tool into the hole marked INT PRESET and depress
the button for 3-5 seconds.
b) Wait 2 minutes, the following should be observed:
i) The Span Filter Indicator should be green (filter is out)
ii) The Detector Status LEDs (located to the left of the display) should be:
(1) Measure: Solid Yellow or Solid Green (typically)
(2) Reference: Solid Green
iii) The display should read 1000 +/- 20 counts and be stable within this allowed range.
iv) If either Detector Status LED is RED, contact CST immediately for assistance.

NOTE: If the display does not fall within the range from 980 to 1020 counts repeat the
calibration step above. Contact CST for technical assistance if three (3) attempts at
calibration are not successful.

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Top of the Transmitter

9) Checking unit operation with the span check neutral density filter.
a) Perform this step with the calibration fluid in place
b) Press and hold the SPAN CHK switch (top of the transmitter) for 1-2 seconds to actuate
the span check assembly. This will move the span check filter into the optical path and
attenuate the fluorescence intensity.
c) The Span Filter Indicator should be red (filter is in).
d) Wait 2 minutes for the signal level to stabilize.
e) The degree of signal attenuation is given in Section 7: Instrument Specifications -
Optical and is listed as the Attenuation Factor (a decimal). The expected signal level is
calculated by taking the Attenuation Factor * Display (or 4-20) value. For example, when
the attenuation factor is 0.407 and the displayed fluorescence level is 997, the expected
display value after inserting the span check filter would be 406 (always round up to the
next whole digit).
f) Press and hold the SPAN CHK switch (top of the transmitter) for 1-2 seconds to actuate
the span check assembly. This will move the span check filter out of the optical path.
g) The Span Filter Indicator should be green (filter is out).
h) Wait 2 minutes for the signal level to stabilize.

NOTE: During span check filter actuation, the displayed values will range from very low
to very high as the assembly moves through the optical path. This is normal operation
for the unit when the span check filter is used.

10) You are now ready to proceed with either lab work, generation of a complete calibration
curve, or process monitoring.

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6.2.2 Mode 1 or Calibrated Operation

Mode 1 operation is the standard or recommended mode of operation for all monitoring
applications. The following procedure assumes that the unit has been installed properly and
has not been moved, altered or otherwise reconfigured since the initial installation. Additionally,
the unit must have been continuously powered with the probe tip mounted in a dark (no ambient
light illuminating the probe tip) environment. If any or all of these conditions have not been met,
please perform the First Power-up Procedure given in Section 6.2.1 (above).

1) Verify that the probe is mounted in a dark environment (the CST calibration bottle, the
process monitoring point, etc.) where ambient light cannot reach the probe tip.
2) Verify that the probe tip is located in a process/test sample. The fluorescence
characteristics of this sample maybe unknown, but CST recommends that the sample be
non-fluorescing. Examples of non-fluorescing samples are:
a) Air,
b) Distilled water, or
c) Non-fluorescing solvents (methanol, cyclohexane, iso-octane, etc.).

Span Filter Indicator

Initialization Button (requires

insertion of the adjustment tool)

Measure Detector Status

Reference Detector Status

3) Add calibration fluid to the probe

mounting point. Wait 2 minutes, the following should be observed:
a) The Span Filter Indicator should be green (filter is out)
b) The Detector Status LEDs (located to the left of the display) should be:
i) Measure: Solid Yellow or Solid Green (typically)
ii) Reference: Solid Green
c) Assuming that the calibration fluid is a relatively close match to your previous calibration
fluid, the display should be reading between 900 and 1100 counts (typically) with a
stability of +/- 20 counts. If this new calibration fluid does not match or has been
intentionally changed since the previous calibration, the display value may not be close
to the expected 1000 count level.
d) If either Detector Status LED is RED, contact CST immediately for assistance.
4) Calibrate the analyzer to your calibration solution. (Mode 1 Operation)
a) Insert the Adjustment/Initialization tool into the hole marked INT PRESET and depress
the button for 3-5 seconds.
b) Wait 2 minutes, the following should be observed:
i) The Span Filter Indicator should be green (filter is out)

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ii) The Detector Status LEDs (located to the left of the display) should be:
(1) Measure: Solid Yellow or Solid Green (typically)
(2) Reference: Solid Green
iii) The display should read 1000 +/- 20 counts and be stable within this allowed range.
iv) If either Detector Status LED is RED, contact CST immediately for assistance.

NOTE: If the display does not fall within the range from 980 to 1020 counts repeat the
calibration step above. Contact CST for technical assistance if three (3) attempts at
calibration are not successful.

Top of the Transmitter

5) Checking unit operation with the span check neutral density filter.
a) Perform this step with the calibration fluid in place
b) Press and hold the SPAN CHK switch (top of the transmitter) for 1-2 seconds to actuate
the span check assembly. This will move the span check filter into the optical path and
attenuate the fluorescence intensity.
c) The Span Filter Indicator should be red (filter is in).
d) Wait 2 minutes for the signal level to stabilize.
e) The degree of signal attenuation is given in Section 7: Instrument Specifications -
Optical and is listed as the Attenuation Factor (a decimal). The expected signal level is
calculated by taking the Attenuation Factor * Display (or 4-20) value. For example, when
the attenuation factor is 0.407 and the displayed fluorescence level is 997, the expected
display value after inserting the span check filter would be 406 (always round up to the
next whole digit).
f) Press and hold the SPAN CHK switch (top of the transmitter) for 1-2 seconds to actuate
the span check assembly. This will move the span check filter out of the optical path.
g) The Span Filter Indicator should be green (filter is out).
h) Wait 2 minutes for the signal level to stabilize.

NOTE: During span check filter actuation, the displayed values will range from very low
to very high as the assembly moves through the optical path. This is normal operation
for the unit when the span check filter is used.

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6) Generate a minimum three (3) point calibration curve for the analyzer. CST recommends a
five (5) point curve. This calibration will allow the operator to generate a predictive equation
that converts the display/4-20 fluorescence intensity (in counts) to the relevant
engineering/application unit. Typically, this will be the fluorophore concentration. The
procedure below can be used as a guide for generating the calibration curve.
a) CST recommends the following four solutions plus the calibration solution for generating
the curve
i) Zero fluorescence – typically the background material
ii) Tenth-span – solution has one-tenth the fluorophore concentration of the calibration
solution
iii) Half-span – solution has one-half the fluorophore concentration of the calibration
solution
iv) One and a Quarter Span – solution has 1.25 times the fluorophore concentration of
the calibration solution.
b) Prepare calibration curve solutions in advance in quantities appropriate for procedure
(this will depend on the size of volume of the probe insertion point).
c) The unit is calibrated and operating in Mode 1.
d) Admit the calibration curve solutions individually and record the fluorescence response
for each sample. Replicate this sequence three times.
e) Take the response data, plot, and then fit the data using either a linear calibration or
second order polynomial equation. CST typically uses the second order polynomial fit,
which provides a better predictor of the fluorescence response for fluorophore
concentrations higher than that of the calibration solution.
f) The analyzer is now fully calibrated and ready for use.

NOTE: The span check filter can be used to provide additional points during instrument
testing and verification. The filter utilized is a neutral density filter and attenuates the
fluorescence response from any sample. Consult the CST QA/QC documentation for the
exact attenuation level in your instrument.

7) You are now ready to proceed with either lab work or process monitoring.

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6.2.3 Mode 2 or Fixed Range Operation

Mode 2 operation is used almost exclusively for application development in the
laboratory. However, this operational mode can also be used for process monitoring for some
fluorophores. CST does not recommend that Mode 2 be used for process monitoring, however.
The following procedure assumes that the analyzer system is operating in Mode 1 within normal
parameters.

1) Verify that the analyzer is operating in Mode 1. If necessary follow the Mode 1 calibration
procedure given in Section 6.2.3.

Span Filter Indicator

Initialization Button (requires

insertion of the adjustment tool)

Measure Detector Status

Reference Detector Status

2) Make sure that the probe tip is immersed in a sample. The fluorescence level of the test
sample need not be known with regard to the Mode 1 calibration curve. Wait 2 minutes, the
following should be observed:
a) The Span Filter Indicator should be green (filter is out)
b) The Detector Status LEDs (located to the left of the display) should be:
i) Measure: Flashing Yellow, Flashing Green, Solid Yellow or Solid Green (dependent
on the fluorophore concentration)
ii) Reference: Solid Green
c) If either Detector Status LED is RED, contact CST immediately for assistance.
3) Initialize the analyzer to the Fixed Range (Mode 2 Operation)
a) Insert the Adjustment/Initialization tool into the hole marked INT PRESET and depress
the button for at least 8 seconds.
b) Wait 2 minutes, the following should be observed:
i) The Span Filter Indicator should be green (filter is out)
ii) The Detector Status LEDs (located to the left of the display) should be:
(1) Measure: Flashing Yellow, Flashing Green, Solid Yellow or Solid Green
(dependent on the fluorophore concentration)
(2) Reference: Solid Green
(3) If either Detector Status LED is RED, contact CST immediately for assistance.
c) The display should read a value between 000 and 1999 counts. If a 1, -1, or -500 is
shown, the unit is off-scale and only Mode 1 may be used to evaluate the fluorophore of
interest in the concentration range desired.

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NOTE: The span check filter can be used to provide additional points during instrument
testing and verification. The filter utilized is a neutral density filter and attenuates the
fluorescence response from any sample. Consult the CST QA/QC documentation for the
exact attenuation level in your instrument.

Top of the Transmitter

4) Checking unit operation with the span check neutral density filter.
a) Perform this step with the calibration fluid in place
b) Press and hold the SPAN CHK switch (top of the transmitter) for 1-2 seconds to actuate
the span check assembly. This will move the span check filter into the optical path and
attenuate the fluorescence intensity.
c) The Span Filter Indicator should be red (filter is in).
d) Wait 2 minutes for the signal level to stabilize.
e) The degree of signal attenuation is given in Section 7: Instrument Specifications -
Optical and is listed as the Attenuation Factor (a decimal). The expected signal level is
calculated by taking the Attenuation Factor * Display (or 4-20) value. For example, when
the attenuation factor is 0.407 and the displayed fluorescence level is 997, the expected
display value after inserting the span check filter would be 406 (always round up to the
next whole digit).
f) Press and hold the SPAN CHK switch (top of the transmitter) for 1-2 seconds to actuate
the span check assembly. This will move the span check filter out of the optical path.
g) The Span Filter Indicator should be green (filter is out).
h) Wait 2 minutes for the signal level to stabilize.

NOTE: During span check filter actuation, the displayed values will range from very low
to very high as the assembly moves through the optical path. This is normal operation
for the unit when the span check filter is used.

5) You are now ready to proceed with sample evaluation in Mode 2

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6.2.4 Tips for Easy Operation

Operation Tip 1: The 53F system is designed for continuous 24/7 operation, so leave the
system running whenever possible. Cycling of power frequently will require calibration steps on
a shorter time interval.

Operational Tip 2: The calibration bottle supplied by CST is a common Nalgene™ bottle
supplied by almost every laboratory supply company. If you are using the bottle for calibration
and system validation, purchase a set of bottles from Fisher to insure that cross contamination
between samples does not occur.

Operational Tip 3: The span check filter can be used to provide additional points during
instrument testing and verification. The filter utilized is a neutral density filter and attenuates the
fluorescence response from any sample. Consult the CST QA/QC documentation for the exact
attenuation level in your instrument.

Operational Tip 4: The First Power-Up Following Installation sequence should be completed
anytime the unit has been powered down for an extended time period (> 7 days).

Operational Tip 5: When powering the unit down, always leave the analyzer in Mode 1.
Restarting the analyzer and verifying operation is much easier with Mode 1.

Operational Tip 6: The 53F analyzer will restart after power down or power failures using the
operating parameters stored prior to the power loss. If power failure occurred in Mode 2, it is
likely that upon the restart analyzer operation will be erratic and unstable. Simply follow the
Mode 2 operation setup (Section 6.2.3) before continuing work.

Operational Tip 7: CST recommends that any sample to be used for Mode 1 calibration have
at least 100 counts of fluorescence intensity in Mode 2.

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6.3 Diagnostics and Troubleshooting

The primary diagnostic features of the 53F are the indicator LEDs on the front panel of
the analyzer and the digital display. This section is a summary of diagnostic information
scattered throughout the manual as appropriate to the section.

6.3.1 Diagnostic Indicators

Span Filter Indicator

Measure Detector Status

Reference Detector Status

Description Function/Use/Operational Information

Indicates position of the span check filter:
Span Check LED
Red = In the optical path (check/verify operation)
Indicator
Green = Out of the optical path (standard operation)
Indicates the amount of energy impinging the fluorescence
emission detector

Monitoring States:
Measure Detector RED = too much light contact CST
Status Indicator Solid Green – acceptable
Solid Yellow – acceptable
Flashing Green – acceptable
Flashing Yellow – acceptable
Indicates the amount of energy impinging the reference
(excitation) detector

Monitoring States:
Reference Detector
RED = too much light intensity contact CST
Status Indicator
Solid Green – acceptable
Solid Yellow – low light intensity contact CST
Flashing Green – low light intensity contact CST
Flashing Yellow – low light intensity contact CST
Normal Range: 000 to 1999

Display Values indicating Failed Operation:

Digital Display
1 in the left most digit
-1 in the left most digit
-500

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6.3.2 Basic Troubleshooting

Span Filter Indicator

Measure Detector Status

Reference Detector Status

TB 2 Connector TB 1 Connector

External Span Control Grounding

Jumper (shown) Terminal

Example of terminal strip connectivity on panel mounted systems.

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Problem: Red LED on MSR DET STATUS or REF DET STATUS

Solution: Power unit off and call CST for a Return Material Authorization (RMA) number.
This unit has too much light impinging the detector or detectors and needs to be
evaluated at the factory.
=====================================================================
Problem: The Span Filter Indicator is neither RED nor GREEN, the unit will not initialize
and there is a humming/buzzing sound.

Cause: There is not a jumper or connection between the COM and SPAN N.C. positions
on the TB2 connector or the common terminal. Therefore, the span assembly is
continually operating and the instrument will not initialize. Failure to have the
connection in place will prevent the instrument from initializing.

Solution: Make sure there is a jumper between the COM and SPAN N.C. positions on the
TB2 connector or terminal strip. For panelized systems with a terminal strip,
there must be a jumper or connectivity to an external/remote Form B contact
(normally closed) switch or relay.
=====================================================================
Problem: The Span Filter Indicator is neither RED nor GREEN and the unit will not
initialize. No sound is heard other than flashing of the lamp.

Cause: The transmitter optical assembly has been shoved up into the transmitter and the
span assembly cannot actuate.

Solution: Call CST for an RMA number. The unit must be repaired by CST.

=====================================================================
Problem: There is a solid yellow indicator on the REF Status LED
Cause: The light passing to the reference PMT does match the factory levels.

Solution: Step 1 - Make sure that the fiber connections to the light source and transmitter
are tight (finger tight). If no change, then:
Step 2 – (Integrated Probe) Make sure that the fiber bundle is connected to the
light source and that the single fiber is connected to the transmitter. (Stand Alone
Probe) Make sure that the port marked “I” on the probe has a fiber running to the
light source, and the port marked “O” has a fiber run to the transmitter. If no
change, then:
Step 3 – Power off the analyzer. Remove the probe from the insertion location
and disconnect the integrated probe (or the stand alone probe run fibers) from
the light source and transmitter.
Step 4 – (Integrated Probe) Hold a flashlight at the end of the probe, and
examine the light transmitted through the fibers to the end of the assembly.
There should be a very bright light emitting from the SMA connections. If the
output is not bright, then call CST for an RMA number, the probe must be
returned for evaluation. (Stand Alone Probe) Disconnect the run fibers from the
probe. Hold a flashlight at the end of the probe, and examine the light
transmitted through the SMA ports at the top of the probe. If the output is not

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bright, then call CST for an RMA number, the probe must be returned for
evaluation. Next, place the flashlight at one end of a fiber optic cable and
examine the light transmitted through the fiber. Repeat procedure for the other
fiber. If either fiber has low transmission, contact CST for the purchase of a new
fiber.
Step 5 – if the probe (or probe + run fibers) all have acceptable transmission,
then reconnect to the analyzer and power the system. Wait 15 minutes. If there
is still no change to the LED color, contact CST for an RMA.

=====================================================================

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Section 7: System Specifications

This section provides the detailed specifications for the 53F analyzer system purchased.
In addition, this section may contain application specific notes on operability, functionality, etc.
Where multiple options are available, the purchased option will be highlighted in yellow.

7.1 Optical Configuration

The optical configuration listed is for a complete analyzer system. The excitation
wavelength selection filter listed is located within the light source SMA connection assembly.
The detectors, reference filter, emission filter, and span check filter specified are internal to the
transmitter housing. The only filter that is field replaceable is the excitation filter.

Optical Method: Fluorescence

Fluorescence Accessory: CST Front Surface Fluorescence Probe

Part Number: 3502

CST 90º Fluorescence Flow Cell

Part Number: 5016

Excitation Light Source: CST Xenon Flash Lamp

Part Number: UVF-100

CST LED Flash Assembly

Part Number:

Excitation Selection Filter: CWL =

Bandpass =
Tmax =

Reference Selection Filter: CWL =

Bandpass =
Tmax =

Emission Selection Filter: CWL =

Bandpass =
Tmax =

Cut-on Wavelength =
Tmax =

Span Check Filter: Type: Neutral Density

Average Optical Density =
Optical Density @ Emission CWL =
Attenuation Factor =

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Detectors: PMT
Silicon Photodiode
InGaS Photodiode

7.2 Transmitter Specifications

Transmitter Model Number:

Transmitter Serial Number:

Operation Program Version: 18.1

20.1
20.2

Span Check Filter: Local and Remote Operation

Optical Connectivity: SMA905

Mechanical
Width: 4.4” (112 mm)
Height: 9.5” (242 mm)
Depth: 3.5” (89 mm)
Weight: <3 lbs (1.4 kg)

Operating Temperature: 41 to 122º F (5 – 50º C)

Storage Temperature: -4 to 122º F (-20 to 50º C)

Display 3½ Digits

Measurement Range: 000 to 1999 counts

Typical Response Time: ~2 minutes to 90% of step change
Maximum Zero Shift: < 2 % of full scale (< 40 counts)
Long Term Output Drift < 5 % per month of full scale (<100 counts)
Repeatability ±1% of full scale (± 20 counts)

Power Requirement 24VDC (18-36 VDC)

Maximum Power Consumption: 1.0 A @ 24 VDC
Typical Power Consumption: 500 mA @ 24 VDC
Analog Output 4-20mA isolated (sourcing)
Analog Loop Resistance 400 Ohms, maximum @ 24 VDC

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7.3 Light Source Specifications

The specifications for each light source are listed separately for clarity. Reference the
highlighted source in Section 7.1 to determine the appropriate specifications.

7.3.1 CST UVF-100 Xenon Flashlamp

Dimensions:
Width: 1.8” (46 mm)
Height: 5.4” (138 mm)
Depth: 3.3” (84 mm)
Weight: < 1 lb (0.45 kg)

Optical Connectivity: SMA905

Spectral Range: 160 – 5000 nm

Stability (CV): <3%
Lifetime: > 1x108 flashes

Voltage Input: 11 – 28 VDC

Typical Input DC Current: 0.2 amps RMS
Peak Input Current: 1.0 amps
Trigger: TTL
Input Connector: 9-pin sub-D

Output Voltage: 400 – 600 VDC (set by CST)

Output Power: 2 watts max
Flash rate: 50 Hz

Operating Temperature: 32 to 104º F (0 – 40º C)

Storage Temperature: -40 to 194º F (-40 to 90º C)
Operating Humidity: up to 95% RH, non-condensing
Shock and Vibration: 1.5G, 5 – 200 Hz (Mil-STD-810C)

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7.4 Fluorescence Accessory Specifications

Please reference section 7.1 Optical Configuration for the highlighted sample accessory.

7.4.1 Integrated Front Surface Probe Specifications

Material of Construction: 316 L Stainless Steel
Titanium
Hastelloy C
Other,

Window Sealing Material: Viton

Kalrez™
Chemraz™

Optical Window Material: UV Grade Sapphire

Optical Connectivity: SMA 905

Pressure Rating: up to 2,000 psig

Temperature Rating: up to 392º F (up to 200º C)

up to 450º F (up to 232º C)
up to 600º F (up to 315º C)

Physical Dimensions
Probe Insertion Length:
Probe Overall Length:
Run Length (Fiber Portion):
Total Assembly Length:
Probe Diameter: 12.0 mm (0.472”)
12.7 mm (0.500”)

7.4.2 Stand-Alone Front Surface Probe Specifications

Material of Construction: 316 L Stainless Steel
Titanium
Hastelloy C
Other,

Window Sealing Material: Viton

Kalrez™
Chemraz™

Optical Window Material: UV Grade Sapphire

Optical Connectivity: SMA 905

Pressure Rating: up to 2,000 psig

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Temperature Rating: up to 392º F (up to 200º C)

up to 450º F (up to 232º C)
up to 600º F (up to 315º C)

Physical Dimensions
Probe Insertion Length:
Probe Overall Length:
Run Fiber Length:
Run Fiber Core Diameter: 600 micrometer
800 micrometer
Probe Diameter: 12.0 mm (0.472”)
12.7 mm (0.500”)

7.4.3 90º Fluorescence Flow Cell Specifications

CST Part Number: 5016

Material of Construction: 316 L Stainless Steel

Titanium
Hastelloy C
Other,

Window Sealing Material: Viton

Optical Material: UV Grade Fused Silica

Pressure Rating: up to 250 psig

Temperature Rating: up to 392º F (up to 200º C)

Process Adapter Connections: ½” Swagelok™

Physical Dimensions:
Length (defined as the optical axis): 5.375” (137 mm)
Width (defined as the flow axis): 4.0” (102 mm)
Height (fluorescence axis): 3.625” (92 mm)

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7.5 Application Specific Information, Notes, and Specifications

This section lists all the application specific information related to specifications,
operations, diagnostics, troubleshooting, and third-party integrated equipment. Additionally,
integrated equipment will be supplied with all manufacturer supplied documentation.

7.5.1 Span Check Remote Operation Specifications

For remote operation of the span check filter assembly, please use normally closed
Form B contact switch or relay.

7.5.2 Enclosure Specifications

Physical
Length:
Width:
Height:

Enclosure Type:

Enclosure Material:

Purge Kit Supplied: Yes

No

Area Classification:

7.5.3 Purge Kit Specifications

Manufacturer:

Please reference the Purge Kit Manual and Specifications documentation supplied with
the analyzer system.

7.5.4 Special Operational Procedures

7.5.5 Additional Comments

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Appendix 1: Application Engineering Assistance

This section is provided to enable operators to use the system in the laboratory or
development environment to determine the best practices for using the analyzer system. CST
terms this activity as application development, and offers this service to customers on a contract
basis. However, CST will consult with customers on their internal application development
activities. Please contact either Mike Ponstingl ([email protected]) or Jess V Ford
([email protected]) for assistance. The phone number is 636-305-0666 and the fax
number is 636-305-1096.
The system was supplied with an optical configuration specific to a given fluorescence
excitation and emission pair, which is defined by the fluorophore of interest. However, the
configuration may be applicable to additional fluorophores as well.. Therefore, the information
provided below is only valid when the fluorophore of interest has excitation and emission
characteristics that correspond to the installed optical filters.

A1.1 Beginning Work

The system was supplied by CST in Mode 1 and calibrated using a material with
fluorescence characteristics defined by the installed filter sets. For most systems, a customer
sample was supplied or a test solution specified for determination of optical configuration and
subsequent analyzer setup. Therefore, the analyzer should be ready for application
development work. Simply follow the procedures given in Section 6.1 to insure functionality.
Prepare the samples of interest to be evaluated with the fluorescence analyzer. It is
imperative that the probe be mounted in a dark environment for application development work,
so that ambient light does not affect the measured values. CST recommends that a brown
Nalgene™ bottle (one was supplied with the system) be used as the sample vessel unless a
dedicated sample handling system is being employed. When purchasing the Nalgene™ bottles
make sure to select the bottles that are described as UV or Light resistant. These bottles are
available from all the major laboratory supply companies such as ThermoFisher, VMR, etc.
Work at CST has found that the bottles have a useable life of approximately 4-6 months when
used for application development.
Before beginning your experimental sequences, make sure you understand which
operational Mode is necessary and read the appropriate Sections above. For work on a
multipoint calibration curve, evaluation of response linearity with concentration, or evaluation of
samples similar to calibration material used by CST, Mode 1 operation the starting point.
However, if the sample is not similar to the CST calibration fluid or the concentration range of
the test samples is quite large, it may be necessary to employ Mode 2. Furthermore, if
comparison of fluorescence levels between possible calibration solutions is desired, Mode 2
must be used. Mode 2 allows the relative fluorescence of each sample to be measured under a
fixed set of operational parameters. CST recommends, however, that Mode 1 operation be
used first, whenever possible, and Mode 2 be employed only after Mode 1 has been found to be
insufficient for the work planned.

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A1.2 Mode 1 (Calibrated Analysis)

Table 1 provides an example of a data organization table commonly used by CST during
application development. Series 1 evaluates zero or non-fluorescing materials. Air and distilled
water are always evaluated by CST in this sequence, followed by solvents or process base fluid
samples supplied by the customer. The goal of this section is to determine the baseline of the
measurement. This baseline is a combination of the native fluorescence of the zero materials
plus contribution from optical materials within the sampling accessory. For applications where
the excitation wavelength is less than 280 nm, fluorescence from the sample accessory may
contribute more background response than the base fluids. It is critical to understand that the
baseline level response is a function of the calibration level. If the fluorescence level of the
calibration material changes, the baseline level will also change.
Series 2 measures the relative fluorescence levels of various samples in relation to the
calibration sample. For this test sequence, CST always defines Sample 0 as the calibration
material. This sequence is used to examine linearity of the fluorescence response and for
generation of a calibration curve. Typically, CST examines two approaches for fitting the data
response: 1) linear (y = mx + b) and 2) a second order polynomial fit (y = Ax2 + Bx + C). In
general, CST finds that the second order polynomial provides the best prediction of
fluorescence response.
Series 3 is used exclusively to test the calibration curve and uses samples with
concentrations both within the Series 2 range, and at least two samples above the calibration
concentration. If the R2 value from a regression analysis of the Series 3 data, using the Series 2
calibration curve, does not exceed 0.95 then the Series 3 results are combined with Series 2
and a new calibration curve is generated. In this instance, Series 3 is repeated with a new set
of test samples at different concentrations.
Based on the test sequences above, CST determines if the calibration concentration
must be modified in order to improve measurement range and/or concentration predictability.
The specification of the calibration concentration is critical for reliable process monitoring. CST
measurement philosophy is to set the calibration level to a sample concentration equal to the
maximum expected fluorophore concentration. Process upsets above this level are recorded as
values above the calibration set point for easy identification, and the relative level of the offset is
determined from the calibration curve. An alternate calibration philosophy sets the calibration
level the midpoint of the expected concentration range. In this instance the maximum process
concentration is very near the upper limit of the analyzer range and process upsets are most
likely recorded as off-scale events. Consequently, the magnitude of the upset condition is
unknown and cannot be extrapolated from the calibration curve.

A1.3 Mode 2 (Fixed Range Analysis)

Mode 2 is ideal for comparing a wide variety of samples to determine a potential
calibration sample for Mode 1 analysis. This situation is especially relevant when the process
sample fluorescence is uncertain; typically the case when process samples were not made
available to CST for optical configuration and instrument setup. Additionally, if multiple
fluorophores are responsive to the optical filter configuration, Mode 2 can be used to compare
the relative fluorescence levels. Once again, Table 1 can be useful for organizing the Mode 2
data set.
As described above, Series 1 tests the background level of the zero or non-fluorescing
materials that may be measured by the fluorescence accessory. In Mode 2, Series 2 is used to
compare the fluorescence response of the test samples with regard to their native fluorescence,

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not relative to an arbitrary calibration level as in Mode 1. Finally, Series 3 is not useful in Mode
2, unless multiple fluorophore comparison is necessary.
It is possible to determine a range factor that will take the values recorded in Mode 2 and
convert them into approximate Mode 1 values, relative to a specific calibration sample (i.e. you
must choose one sample as the “calibration” sample). Outlined below are the steps involved:

Step 1: Determine the calibration sample

Step 2: Determine the Range Multiplier

1000
Range Multiplier =
Mode 2 Measured Value

Step 3: Convert the Mode 2 sample value into an approximate Mode 1 value by

Mode 1 Prediction = Mode 2 Value * Range Multiplier

CST recommends that only samples with at least 100 counts of fluorescence in Mode 2
be used as calibration fluids in Mode 1. If this guideline is not followed, measurement instability
may result.

A1.4 Useful Equations

1: Converting the 4 – 20 output to Fluorescence Intensity (counts)

Let x = 4-20 output

⎛ x−4⎞
⎜ ⎟ *1999 = Counts
⎝ 16 ⎠

2: Converting the 4 – 20 output to Engineering Units

Let x = 4-20 output

Let y = full scale range of the engineering unit

⎛ x−4⎞
⎜ ⎟ * y = Engineering Value
⎝ 16 ⎠

3: Calculating the Attenuation Factor from the test data

Let x = measured value

Let y = measured value with the span filter inserted

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y
Attenuating Factor =
x

Table 1: Data organization example.

Operational Mode:

Series 1: Base Fluid Fluorescence Levels.

Material Probe/Analyzer Fluorescence Span Filter Fluorescence

Series 2: Process Sample Analysis

Sample Number Analyte Concentration Measured Fluorescence
0
1
2
3
4
5
6
Series 3: Monitoring verification using selected calibration (initialization) procedure
Sample Number Analyte Concentration Measured Fluorescence
0
1
2
3
4
5
6

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Appendix 2: Advanced Troubleshooting

A2.1 Emergency Troubleshooting Procedure

This procedure should be used to test the operation of the UV PhotoX transmitter
electronics in the field. This procedure should only be utilized by CST trained service personnel
and only at the request of a CST Technical Staff member. Connect the probe to the analyzer
and place the probe tip in the calibration bottle.

The following steps must be completed in sequence to test the electronics:

1) Turn the transmitter on and allow the system to self-initialize: ~10-15 seconds
2) Press and hold the INT button
3) While holding the INT button, turn off the power, then turn the power back on
4) Release the INT button
a) MSR LED should be solid green
b) REF LED should be off (no color)
c) Display = 000
d) 4-20 output = 4 mA
e) Record all deviations from the information above
5) Press the INT and hold until
a) MSR LED should be solid red
b) REF LED should be off (no color)
c) Display = 1900
d) 4-20 output = 19.2 mA
e) Record all deviations from the information above
6) If the outputs match steps 4 and 5, then the problems are in the optical portions of the
analyzer. Power the unit off
7) If the outputs do not match steps 4 and 5, call CST with the outputs obtained in steps 4 and
5. Power the unit OFF

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Appendix 3: Analyzer System Engineering Drawings

See attached.

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Appendix 4: QA/QC Documentation

See attached.

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