Food & Beverage

ANALYTICAL METHODS MANUAL

TABLE OF CONTENTS
1–3 Introduction Food Analysis Simplified
4–6 Application Note 200 Simultaneous Measurement of L-Lactate and Ethanol in Tomato-Based Products

7 – 8 Application Note 201 Ethanol Determination in Beer

9 Application Note 202 J. Lohr Winery Utilizes YSI Instruments in Managing Dissolved Oxygen

10 – 11 Application Note 203 Choline Determination

12 – 14 Application Note 204 Simultaneous Measurement of Glucose and Sucrose Utilizing External Hydrolysis

15 – 16 Application Note 205 Simultaneous Measurement of Glucose and Sucrose in Peanut Butter

17 Application Note 206 Lactose Measurement in Cheese

18 – 19 Application Note 207 L-Glutamate Determination

20 Application Note 208 Determination of Hydrogen Peroxide

21 – 22 Application Note 209 Simultaneous Measurement of Glucose and Sucrose in Frozen Ice Cream Bars

23 – 24 Application Note 210 Glucose Measurement in Canned Green Beans

25 – 26 Application Note 211 Glucose Measurement in Frozen Green Beans

27 – 28 Application Note 212 L-Lactate in Lunch Meats

29 - 30 Application Note 213 Simultaneous Measurement of Glucose and Sucrose in Corn and Peas

31 – 32 Application Note 214 Simultaneous Measurement of Glucose and Sucrose in Cereal Products

33 – 34 Application Note 215 Simultaneous Measurement of Glucose and Sucrose in Baked Goods

35 – 36 Application Note 216 Simultaneous Measurement of Glucose and Sucrose in Sweetened Condensed Milk

37 – 38 Application Note 217 Glucose Measurement in Corn Syrup and Other Syrup Products

39 – 40 Application Note 218 Measurement of Glucose and Sucrose in Potatoes

41 – 42 Application Note 219 Dextrose Measurement in Potatoes

43 – 44 Application Note 220 Simultaneous Measurement of Dextrose and Sucrose in Molasses

45 – 46 Application Note 221 Sucrose Measurement in Molasses

47 – 48 Application Note 222 Determination of % Cook in Extruded Cereal Products

49 – 51 Application Note 223 Determination of % Cook in Extruded Cereal Products Using Chemical Solubilization
Food Analysis
SIMPL IFIED
William Miller
YSI Life Sciences Product Manager

There is a science to flavor. The field of food science YSI Biosensor Technology Advantage
has evolved significantly to meet the growing demands YSI’s innovative enzyme electrode technology harnesses
for food composition and characteristic determination. the power of enzymatic specificity and catalysis to pro-
Trends and demands of consumers, the food industry, vide a rapid, precise analytical tool. Enzymes, which are
and national and international regulations challenge powerful biological catalysts, can accelerate reactions by
food scientists as they work to monitor food composition, factors of at least one million2. Additionally, enzymes are
authentication and to ensure the quality and safety of the highly specific for both the reaction catalyzed as well as
food supply1. its choice of substrate2. YSI immobilizes substrate-specific
enzymes between two selective interfering membranes
For example, regulatory compliance with the US Food and couples the membrane component to a platinum
& Drug Administration’s (FDA) Nutrition Labeling and electrode, which results in a highly specific measurement
Education Act and other country-specific regulatory for a given substrate.
requirements, mandate disclosure and labelling of food
ingredients and their respective quantities. Also, more YSI Life Sciences’ family of Biochemistry Analyzers can
consumers are interested in food product nutrient
content, so that they may make informed purchase
meet demands through automated, plug-and-play analytical
decisions based on diet and health. solutions for raw materials testing, food R &D, in-process
monitoring and final product testing – especially when
Lastly, product quality management and competitive
pricing are driving industry trends in analytical methods rapid, precise and economical analysis of saccharides,
and technologies. Analytical methods must be applied amino/organic acids, alcohols and electrolytes are required.
across the entire food supply chain to achieve the
desired final product quality1. Cost reduction initiatives
and workforce downsizing mandate the implementation
of economical, easy-to-use technologies. Thus, simple,
costeffective, standardized technology platforms,
providing rapid, accurate results, are of the essence
for today’s food and beverage space.

Achieving these industry requirements can be a chal-

lenge. However, YSI Life Sciences’ family of Biochemistry
Analyzers can meet these demands through automated,
plug-and-play analytical solutions for raw materials
testing, food R&D, in-process monitoring and final
product testing – especially when rapid, precise and
economical analysis of saccharides, amino/organic
acids, alcohols and electrolytes are required.

1 YSI Life Sciences | Introduction

 To better explain how the enzyme electrode technology
works, an example of a common food ingredient analysis,
i.e., dextrose (D-glucose), will be used. An enzyme probe
is fitted with a three-layer membrane containing immo-
bilized glucose oxidase enzyme in the middle layer. The
face of the probe, covered by the membrane, is situated
in a buffer-filled sample module into which the dextrose
sample is injected by the instrument. The dextrose
diffuses through the first membrane. When it contacts
the immobilized glucose oxidase, it is rapidly oxidized,
producing hydrogen peroxide (H2O2).
Figure 1
YSI Enzyme Electrode Technology. Enzyme specificity and
H2O2, in turn, passes through the inner membrane and
proprietary membrane attributes allow for rapid, precise
is oxidized at the platinum anode, producing electrons.
and essentially interference-free analysis.
A dynamic equilibrium is achieved when the rate of
H2O2 production and the rate at which H2O2 leaves the
immobilized enzyme layer are constant and is indicated
by a steady state response. The electron flow is linearly
proportional to the steady state H2O2 concentration and,
therefore, to the concentration of the substrate. All of this
occurs in a matter of seconds, which results in a sample
analysis cycle of 60 seconds or less per chemistry
analyzed. See Figure 1.

Sample analysis requires little or no preparation due to

the YSI Biochemistry Analyzer’s unique sample chamber
design and membrane characteristics, which makes the
enzyme electrode impervious to sample color, pH,
turbidity, cell concentrations, salts, proteins, detergents
and other low molecular weight interferences.
Additionally, only small sample volumes are needed,
ranging between 10–50 μl. The YSI 2900 Series
Biochemistry Analyzers are highly flexible, modular
platforms with a range of configurations, options and
accessories to meet the various needs of the food
Figure 2 process cycle, regardless of scale of operations (Figure 2).
The YSI 2950 Biochemistry Analyzer can simultaneously
measure up to six chemistries for raw materials, R&D, Up to six different chemistries can be measured simulta-
in-process and final product sample analysis. neously using automated, high-throughput options, i.e.,

2 YSI Life Sciences | Introduction

96-well plates, stat mode for immediate process sample Conventional methods can be costly and/or time con-
checks and automated on-line sample analysis for the suming. The YSI biosensor technology offers a low cost
food and beverage process and effluent streams. alternative while providing rapid sample analysis due to
Each analyzer offers an intuitive graphic user interface, the characteristics of the immobilized enzyme membrane
onboard training videos and multiple data management technology. Additional time savings can be realized as
options. As compared to other conventional methods, no sample preparation is required in most cases. For
such as HPLC, YSI Biochemistry Analyzers offer a low- example, analyzing glucose and sucrose in high salt food
cost alternative for both initial capital investment and samples is easily achieved with no sample preparation.
cost per chemistry test ($0.10 - $0.70 USD/sample). In contrast, trying to perform this type of analysis using
Other features include low maintenance requirements HPLC is difficult, and sometimes impossible, due to high
and easy product changeover. salt interference. Other food and beverage applications in
which the YSI enzyme electrode technology has replaced
Industry Applications other analytical methods include various analyte monitor-
For over two decades YSI’s Biochemistry Analyzers ing of ice cream products, beer production, sugar-free
have been the analytical technology of choice for many food and beverages, dry bakery mixes and animal food.
major food and beverage manufacturers. The operations
in which the YSI biosensor technology has been utilized As most of this discussion has implied at-line or off-line
spans from raw materials analysis through final product laboratory use, YSI Biochemistry Analyzers have been
QC testing and effluent monitoring. The table below lists successfully employed for on-line process monitoring.
some of the food and beverage applications in which YSI With industry trends moving towards automated, online
Biochemistry Analyzers have been employed. and inline monitoring of food and beverage processes,
real-time monitoring of food ingredients and effluent
Table 1 can be achieved with YSI’s online monitoring and control
Examples of YSI End-User Current system. This bolt-on function automatically draws a fluid
and Emerging Applications sample from the process stream and delivers the sample
directly to the YSI Biochemistry Analyzer. The sampling
Analytical Application Function is performed aseptically, and the online analytical results
Choline in infant formula R&D, In-process can be communicated immediately to your SCADA or
process management system for real-time data acquisi-
Choline in animal feeds Final product
tion and/or feedback control.
Dextrose, Sucrose & Lactose in candy Final product
Dextrose & Sucrose in cereal R&D, In-process In conclusion, food industry trends, regulatory require-
ments and needed analytical improvements dictate the
Dextrose - starch-to-glucose conversion R&D, In-process requirement for simplified, low-cost, rapid and precise
Dextrose & Sucrose in potatoes/french fries Raw materials, In-process analysis of food and beverage ingredients throughout the
entire process train. These attributes should be coupled
Dextrose & Lactate in wine production In-process
with versatile, scale-independent technologies to provide
Glutamate (MSG) in broth & food bases Final product standardized, plug-and-play analytical platforms which
Lactose in cheese filtration R&D span across the food and beverage cycle.

Lactose in low lactose milk product In-process YSI’s unique enzyme biosensor technology helps satisfy
Lactate in tomato-based products In-process, Final product this need for common food ingredients including mono-
and disaccharides, alcohols, amino acids and more.
Sucrose content in soft drinks In-process
If you would like to learn more about YSI’s food and
beverage analytical solutions, please contact us at
ysi.com/lifesciences, [email protected] or 800-765-4974. n

References
[1] S. Suzanne Nielsen, ed. Food Analysis. New York: Springer, 2010.
[2] Stryer, Lubert. Biochemistry. New York: W.H Freeman
and Company, 2000.

3 YSI Life Sciences | Introduction

 Application Note 200LS

Simultaneous Measurement of L-Lactate and Ethanol

in Tomato-Based Products
INTRODUCTION
L-lactate and ethanol concentrations in complex matrices Precision of replicate samples was determined from
such as tomato paste and ketchup can be measured in about selected samples; and percent recovery was determined
one minute with minimal preparation using a YSI Biochemistry for samples spiked with both lactate and ethanol. The
Analyzer. YSI’s unique enzyme electrode technology provides material and method sections are written to demonstrate
for specific lactate measurements in the range of 10 to 1335 one approach to measuring lactate and ethanol in a tomato
ppm (mg/L). Ethanol can be measured in the range of 20 to process application. The results section demonstrates typical
1000 ppm. Measurements are virtually unaffected by color, precision and accuracy when using a YSI Biochemistry
turbidity, density, pH, or the presence of chemical substances. Analyzer in process applications.

When configured with the YSI 2329 lactate oxidase

membrane, a YSI 2786 alcohol oxidase membrane and
YSI 1579 buffer, the YSI analyzer simultaneously measures
lactate and ethanol after aspiration of just 15 microliters
of sample. Samples may require centrifugation to remove
particulates; therefore the lactate and ethanol are read from
the supernatant. Lactate concentrations that exceed 1335
ppm and ethanol concentrations that exceed 1000 ppm
may require dilution. Results are displayed and printed. The
sample automatically flushes from the electrode chamber 45
seconds after results and the YSI analyzer is ready to measure
the next sample. Turn around time is under two minutes.

In the manufacture and packaging of ketchup and

related tomato products, both lactate and ethanol Enzymatic oxidase
Analyte + O2 H2O2 + By product
concentrations can be used to quantify microbial testing
of ingredients, in-process and finished products. Lactate-
Oxidized at probe
producing bacteria and ethanol-producing yeast may H2O2 O2 + 2H+ + 2e-
both contribute to microbial load. When mixed with broth +0.7V
and grown in cultures for 1-5 days, the lactate and ethanol
concentrations may be correlated with the degree of Substrate proportional to electron flow...
microbial load. This method of measuring lactate and/or Substrate 2e-
ethanol typically reduces test time by hours and provides
results that better predict potential flavor issues and incipient
spoilage compared to traditional microbial methods.1

Data in this study supports the feasibility of simultaneously

measuring L-lactate and ethanol in tomato-based products.
Lactate and ethanol were measured in supernatants
of a diluted ketchup sample that was obtained from
a commercially available source. In order to prove
feasibility ketchup samples were spiked with lactate and
ethanol to levels that would be typical of changes seen in
microbial load tests (50 to 150 ppm range for both lactate
and ethanol). YSI 2900 YSI 2950

4 YSI Life Sciences | Application Note 200LS

 Application Note 200LS

METHODS
A sample from commercially available ketchup was collected Following protocol described in the previous paragraph,
and diluted 1:1 by volume with reagent water to reduce 4.000 ml of lactate and 4.000 ml of ethanol were combined
viscosity. Two aliquots of the sample were collected and with 92 ml of diluted ketchup. The theoretical changes were
transferred to 1.5 ml plastic ‘eppy’ tubes, and then spun determined to be 106.8 ppm and 128.0 ppm above base
by centrifuge. The supernatants were presented to the YSI for lactate and ethanol, respectively.
Biochemistry Analyzer for ten (10) measurements each of
L-lactate and ethanol. The readings were recorded and the RESULTS
precision of each analyte was determined. The final base YSI Biochemistry Analyzer Precision
lactate and ethanol readings were averaged and used for Unspiked Samples
to calculate spike/recovery values in the second study. Selected samples of diluted ketchup (1:1) were used for
precision studies. Ten (10) replicates of each sample were
Since levels of lactate as low as 50 ppm (mg/L) can indicate performed. Results are shown in tables on page 6 for lactate
potential flavor/spoilage issues in tomato products during and for ethanol.
microbial load tests as determined by human taste testers
(personal communication, HJ Heinz), and changes in lactate The standard deviation (STD) was determined for each
levels in cultures of approximately 100 ppm can indicate replicate series. Imprecision was no greater than 2.25%
incipient spoilage, the spike/recovery tests were designed expressed as CV.
with this in mind. Ethanol levels have been less studied,
however changes in the 50 to 150 ppm range represents Percent Recovery of Spiked Samples
a reasonable change to detect yeast or mold effects in the As an evaluation of measurement accuracy, spiked samples
microbial load tests. were measured using the YSI Biochemistry Analyzer.
The spiked sample values were then compared to the
calculated spiked sample value. To clarify the process, a
sample of known concentration, referred to as the unspiked
sample, is combined with a spike of known concentration
to create the spiked sample. The expected value of the
spiked sample is listed in the table (pg 6) as the “Calculated”
value. The value of the spiked sample obtained with the YSI
Biochemistry Analyzer is then designated the “Spiked” value.

Spiked samples of diluted ketchup were measured using

the YSI Biochemistry Analyzer within 30 minutes of spiking
and mixing. Aliquots were each measured in triplicate and
YSI 1530 (30 mmol/L; 267 mg/dL L-lactate) was used as a the average result was recorded for each. The results are
stock lactate standard. YSI 2790 (3.20 g/L; 320 mg/dL ethanol) shown in Table 1 on page 6.
was used as a stock ethanol standard. Both were prepared
by YSI metrologists with metrology-grade glassware and SUMMARY
weights using the highest purity standards available. Experimental results shown above demonstrate that the YSI
Biochemistry Analyzer can simultaneously measure lactate
Into a 100 ml volumetric cylinder 2.000 ml of lactate standard and ethanol in a tomato matrix with adequate precision and
and 2.000 ml of ethanol standard were combined with accuracy to make process and quality assurance decisions
96 ml of ketchup that had been diluted 1:1 with reagent in tomato product manufacturing. Although the methods
water. The additions represent 53.4 mg of lactate and section describes a protocol appropriate for screening
64.0 mg of ethanol. These values represent theoretical potential flavor issues and incipient spoilage in finished
changes of 53.4 ppm and 64.0 ppm increases above product, similar approaches provide means to test microbial
base lactate and ethanol concentrations, respectively load in ingredients or to assess sanitation needs in process
after corrected for volume (0.96 x unspiked conc.). equipment. L-lactate is useful in identifying growth of lactic
acid bacteria while ethanol identifies the potential presence
of yeasts and molds.
5 YSI Life Sciences | Application Note 200LS
 Application Note 200LS

L-LACTATE
Mean
Sample Replicates STD ppm CV (%)
ppm
LAC-B1 10 72.5 1.28 1.76%
LAC-B2 10 72.8 1.64 2.25%

ETHANOL
Mean
Sample Replicates STD ppm CV (%)
ppm
ETH-B1 10 181.7 3.97 2.19%
ETH-B2 10 181.2 3.85 2.13%

ORDERING INFORMATION
Table 1 YSI Part Numbers:
Calcu- 2900 Biochemistry Analyzer
Sample Spike* Unspiked* Spiked* Recovery
lated* 2328 Lactate Linearity Test Standard (15.0mmol/L)
LAC-1 53.4 72.6 122.3 123.1 99.4% 2329 L-Lactate Oxidase Membrane Kit
2363 Potassium Ferrocyanide Test Solution
LAC-2 106.8 72.6 175.3 173.6 101.0% 2392 NaCl Solution (for membrane installation)
2776 Glucose/Lactate Calibrator (0.50 g/L lactate)
ETH-1 64.0 181.5 244.8 238.2 102.8%
2786 Alcohol Oxidase Membrane Kit
ETH-2 128.0 181.5 307.2 295.0 104.1% 1579 Buffer Kit
2792 Low Concentration Ethanol Kit (0.50 g/L; 1.00 g/L)
*All values in ppm or mg/L units - 2759 0.50 g/L Ethanol Standard
- 2769 1.0 g/L Ethanol Standard n

1
YSI greatly appreciates the contributions of John Palombi, Laura Bautista and Phil Vendemio (all of Heinz North America) who provided
valuable insight and understanding regarding quality control in tomato product processing.

6 YSI Life Sciences | Application Note 200LS

 Application Note 201LS

Ethanol Determination in Beer

INTRODUCTION
Ethanol concentrations in complex matrices such as beer can YSI 2900
be measured directly and quickly using any YSI 2900 Series
Biochemistry Analyzer. YSI’s unique biosensor technology
provides for rapid ethanol measurement. Measurements
are virtually unaffected by color, turbidity, density, or pH.

When a sample is injected into the sample chamber,

the ethanol diffuses into the membrane containing
alcohol oxidase. The ethanol is immediately oxidized to
hydrogen peroxide and acetaldehyde. The hydrogen F. The following instrument setup is recommended:
peroxide is detected amperometrically at the platinum
electrode surface. The current flow at the electrode Probe A Parameters
is directly proportional to the hydrogen peroxide
concentration, and hence to ethanol concentration. Chemistry Ethanol
Unit g/L
Other alcohols can interfere with ethanol measurement Calibrator 2.00 g/L
by also oxidizing and producing a signal. Fortunately, Sample size 15 μL
most alcohols have much lower responses to alcohol End Point 45 Sec
oxidase than ethanol. One notable exception is methanol, Cal Station 21
which is over three times as sensitive to the enzyme as
ethanol. Therefore, samples must be methanol-free. Probe B Parameters

This application note demonstrates how simply and quickly B Chemistry None
ethanol concentrations can be determined using the following
method. Precision of replicate samples was determined from Autocal Parameters
selected samples; and percent recovery was determined
for samples spiked with ethanol. The results section Time (min) 30
demonstrates typical precision and accuracy when using a Sample 5
YSI Biochemistry Analyzer in process applications. T Shift (°C) 1
Cal Shift (%) 2
I. MATERIALS & SETUP
1
Calibrator should be used in small quantities and refreshed frequently
A. YSI 2900 Series Biochemistry Analyzer -
due to evaporation. Calibration should be performed from a test tube.
equipped with a 2786 Ethanol Membrane and
1579 Ethanol Buffer.
II. METHOD
B. 2790 Ethanol standards (2.00 g/L, 3.20 g/L). A. Dilute sample to bring ethanol concentration into
Place the 2.00 g/L calibrator solution in Station 2. the linear range of the instrument, which is 0.04 to
3.20 g/L. Samples with 1.5 to 2.5 g/L ethanol will give
C. Connect the 2900 Series Biochemistry Analyzer to the best results.
a suitable power source.
B. Calibrate the 2900 Series Biochemistry Analyzer with a
D. Perform the instrument and membrane check 2.00 g/L ethanol standard solution.
described in the Operations Manual.
C. Check the linearity of the membrane at least once a
E. Volumetric glassware (Class A recommended). day with an ethanol linearity check solution (3.20 g/L).
Refer to the User’s Manual for specifications.

7 YSI Life Sciences | Application Note 201LS

 Application Note 201LS

D. Assay the sample prepared in A by aspiration into the IV. RESULTS / DISCUSSION
2900 Series Biochemistry Analyzer. The linear range Several beers were analyzed for their ethanol concentrations
of the system is 0.04 to 3.20 g/L ethanol. If the value (w/v) using the YSI biochemistry analyzer and Sigma
reported exceeds this, further dilution is required. Test Kit 332-BT. The YSI analyzer results and Sigma
Test Kit results are compared in the following table:
E. Calibrate frequently as described in the User’s Manual.

III. CALCULATIONS
YSI Sigma
To calculate % ethanol, multiply the reported value by the
appropriate dilution factor. Beer A 4.40% 4.39%

Example: 5.00 mL of beer was diluted to 100 mL in Beer B 3.75 3.76

a Class A volumetric flask. When assayed, the value
Beer C 3.89 3.87
reported was 1.56 g/L ethanol.
Beer D 3.74 3.79
% Ethanol: = 0.03120g ethanol/mL beer
1.56 g/L x
0.100L/5mL = 3.12% (w/v)
YSI’s proprietary immobilized enzyme membrane technology
For a v/v result, divide the mass by the density of ethanol provided accurate ethanol results within one minute of
(0.789 g/mL at 20°C). sampling. The YSI 2900 Biochemistry Analyzer’s ability to
provide rapid, precise analyses makes it ideal for brewing
= 0.0395mL ethanol/mL beer process monitoring and control and QC analysis of beer
0.03120 g/mL /
harvest samples.
0.789 g/mL
= 3.95% (v/v)
ORDERING INFORMATION
YSI Part Numbers:
2900 Biochemistry Analyzer
2786 Ethanol Membrane Kit
2790 Ethanol Standards Kit
1579 Carbonate Buffer Concentrate
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

YSI 2900 Biochemistry Analyzer

8 YSI Life Sciences | Application Note 201LS

 Application Note 202LS

J. Lohr Winery Utilizes YSI Instruments

in Managing Dissolved Oxygen
INTRODUCTION
Although oxygen is a necessary component in the wine and 5010-W wine bottle BOD
aging process, it can also be detrimental to overall wine probe. The results are used for
quality. Too much oxygen can cause browning in white wine Quality Assurance/Quality
wines and induce flavor degradation in both red and white Control. If it is determined
wines. The J. Lohr Winery in San Jose, California, consistently that too much oxygen has
strives to minimize and control oxygen exposure to their been introduced into the wine,
premium wines throughout their winemaking process. corrective measures can be
quickly implemented to rectify
DISSOLVED OXYGEN (DO) DETERMINATION the problem. This type of careful
Using the YSI 550A handheld DO and temperature inspection allows J. Lohr Winery
instrument, the laboratory staff at J. Lohr Winery can quickly to consistently produce high-
and simply check DO levels of wine stored in stainless steel quality wines.
tanks and oak barrels. Obtaining DO concentrations in these
vessels provides a baseline measurement to determine the Today the J. Lohr products are
amount of oxygen introduction of the bottling process when available throughout the United
the wine is moved from one location to another through States and in more than 25 YSI 550A
pumps and hoses. Fittings on tank valves, hoses, and countries worldwide. The goal Dissolved Oxygen
instrument
pumps can be checked as potential sources of the oxygen of J. Lohr Winery is to produce
introduction. Preventative measures, such as nitrogen the finest varietals in the world, using a style that focuses
gas sparging can greatly diminish the amount of oxygen on flavor and complexity through vineyard selection,
introduction during wine transfer. technology and innovation.

Oxygen monitoring after bottling ensures minimal oxygen This goal has led Jerry Lohr, President and Owner, and his
introduction from the bottling tank to a 54-valve filler. The team to develop three tiers of wines produced from estate
YSI 5100 Dissolved Oxygen Instrument is used to determine vineyards: J. Lohr Cuvée Series, J. Lohr Vineyard Series, and
the dissolved oxygen levels and temperature of the wine J. Lohr Estates. In addition, J. Lohr Winery produces three
once it is bottled. These measurements are achieved using tiers of wines to meet the needs of everyday and entry-level
the YSI 5010-W, Wine Bottle BOD Probe, which is specifically wine consumption: Crosspoint Vineyards, Cypress, and
designed to fit directly into the neck of a wine bottle. The Painter Bridge. n
5010-W probe has a tapered fit, which allows a best fit and
seal atop the bottle. The probe also has a self-powered For additional information YSI 5100 and
stirrer attached to its submersible end, which ensures an regarding the J. Lohr Winery, 5010-W measure
optimal flow of wine past the membrane within the small please visit: jlohr.com dissolved oxygen
confines of the bottle. directly in the
wine bottle.
For additional information
WINE PROCESS MONITORING regarding the measurement of
DO levels can be checked at various points in the wine DO at the J. Lohr Winery,
bottling process, such as filler bowl seals and filler spouts. please contact:
DO baseline values are determined from the bottling tank Susan Kanzaki
using the 550A handheld DO instrument. Lab Supervisor, J. Lohr Winery

Bottles are then pulled from the bottling line after the corker
and checked immediately in the laboratory using the 5100

9 Application Note 202LS | YSI Life Sciences

 Application Note 203LS

Choline Determination
INTRODUCTION II. METHOD
Choline concentrations in complex matrices can be A. Total choline concentration should not exceed 450
measured directly and quickly using the YSI 2900 Series mg/L, as determined on Part D below; otherwise the
Biochemistry Analyzer. YSI’s unique enzyme technology sample will require further dilution. Use volumetric
provides for specific choline measurement. Measurements glassware for all dilutions. Dilute with either water or
are virtually unaffected by color, turbidity, density, pH, or the 2357 buffer.
presence of reducing substances.
B. Calibrate the 2900 series instrument with a 175 mg/L
When a sample is injected into the sample chamber, the Calibration Standard.
choline diffuses into the membrane containing choline
oxidase. The choline is immediately oxidized to hydrogen C. Check the linearity of the membrane at least once a
peroxide and betaine. The hydrogen peroxide is detected day by injection of a choline linearity check solution
amperometrically at the platinum electrode surface. The (450 mg/L). Refer to the User’s Manual (Section 5) for
current flow at the electrode is directly proportional to the specifications.
hydrogen peroxide concentration, and hence to choline
concentration. D. Assay the sample by aspiration into the 2900 series
instrument. The linear range of the system is 5 to 450
I. MATERIALS & SETUP mg/L choline. If the value reported exceeds this, further
A. YSI 2900 Series Biochemistry Analyzer - equipped dilution is required.
with a 2771 Choline Membrane and 2357 Buffer.
E. Calibrate frequently as described in the User’s
B. Choline standards (175 mg/L, 450 mg/L). Manual.

C. Connect the 2900 Series instrument to a suitable III. CALCULATIONS

power source. To calculate % choline, multiply the reported value by the
appropriate dilution factor.
D. Perform the instrument and membrane daily checks
described in the User’s Manual. Example: 5.0 grams of pet food and 100 mL of water were
mixed in a blender for 5 minutes. The supernatant was
E. Volumetric glassware (Class A recommended). analyzed for choline. The value reported was 77.0 mg/L
choline.
F. The following instrument setup is recommended.
Sample size: 25 μL % Choline: = 0.0015 g choline/g pet food
77.0mg/L x 0.100
Probe A Parameters Probe B Parameters L/5000 mg = 0.15% (w/w)

Chemistry Choline B Chemistry None

Unit mg/L (ppm)
Calibrator 175 mg/L Autocal Parameters
End Point 30 Sec Time (min) 30
Sample 2
T Shift (°C) 1
Cal Shift (%) 2

10 Application Note 203LS | YSI Life Sciences

 Application Note 203LS

Example: Infant formula was aspirated into the 2900 Series ORDERING INFORMATION
(no dilution). The choline content was as follows: YSI Part Numbers:
2900 Biochemistry Analyzer
Sample mg/L Choline 2771 Choline Membrane Kit
Infant Formula A 232 2772 Choline Standard Solution (175 mg/L)
2773 Choline Standard Solution (450 mg/L)
Infant Formula B 138
2357 Buffer Kit
Medical Nutritional Formula A 398 2363 Potassium Ferrocyanide Test Solution
Medical Nutritional Formula B 386 2392 NaCl Solution (for membrane installation) n

11 YSI Life Sciences | Application Note 203LS

 Application Note 204LS

Simultaneous Measurement of Glucose and Sucrose

Utilizing External Hydrolysis
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in F. Perform the instrument and membrane check
complex matrices can be measured directly and quickly described in the Operations Manual.
using the YSI 2900 Series Biochemistry Analyzer. YSI’s unique
enzyme technology provides for rapid glucose and sucrose G. Volumetric glassware (Class A recommended).
measurement. Measurements are virtually unaffected by
color, turbidity, density, pH, or the presence of reducing H. The following instrument setup is recommended:
substances. Sample Size 25 μL

When a YSI 2900 Series Biochemistry Analyzer is equipped Probe A Parameters

with a glucose membrane, both sucrose and glucose Chemistry Glucose
concentrations can be measured. This is accomplished by Unit g/L
first determining the glucose concentration. The sucrose Calibrator 1.80 g/L
is then converted to glucose, and the total glucose End Point 30 Sec
concentration is measured. The difference in the responses
corresponds to the sucrose concentration. Autocal Parameters
Temperature 1°C
After a sample is injected into the sample chamber, the
Time 30 Min
glucose diffuses to the glucose membrane, which contains
Sample 5 Sam
glucose oxidase, and is oxidized to hydrogen peroxide and
Cal Shift 2%
D-glucono-δ-lactone. The hydrogen peroxide is detected
amperometrically at the platinum electrode surface. The
current produced is directly proportional to the hydrogen II. METHOD
peroxide and glucose concentrations. In this section two different case studies will be described.
These examples can be followed when doing analysis on
For more information on this system, refer to the products that may contain similar concentrations of glucose
Operations Manual. and sucrose.

Case #1
I. MATERIALS & SETUP Example: The product is a powdered seasoning mix that is
A. YSI 2900 Series Biochemistry Analyzer - equipped believed to contain 14% glucose and 2% sucrose. The fol-
with a 2365 Glucose Membrane and 2357 Buffer. lowing sample preparation was used:
A. Weigh out about 10 grams of the powder (record
B. Glucose (1.80 g/L) standard solution.
exact weight).
C. Buffer Diluent (40 g/L NaH 2PO 4, 10 g/L Na 2HPO 4
B. Transfer the powder to a 100 mL volumetric flask
in reagent water).
using buffer diluent to rinse and dilute. Fill the flask
D. Invertase - Sigma Chemical Company I-4504 to the mark with buffer and mix. Allow the solution to
recommended. equilibrate for about 20 minutes.

E. Connect the 2900 Series instrument to a suitable C. Remove about 3 mL of the solution in B and add ~2
power source. mg of invertase enzyme. Stir gently until dissolved.
Cover the sample and allow to incubate at room
temperature for 20 minutes.

12 YSI Life Sciences | Application Note 204LS

 Application Note 204LS

D. Calibrate the 2900 Series with 1.80 g/L glucose F. Check the linearity of the membrane at least once a
standard solution. day by injection of an appropriate linearity standard.
Refer to the Operations Manual for specifications.
E. Check the linearity of the membrane at least once
a day by injection of an appropriate linearity standard. G. Assay the sample prepared in B by aspiration into
Refer to the Operations Manual for specifications. the 2900 Series instrument. This is the free glucose
concentration (Dfree).
F. Assay the sample prepared in B by aspiration into
the 2900 Series instrument. This is the free glucose H. Assay the sample prepared in C (with invertase).
concentration (Dfree). The value reported is the sum of the free glucose and
that produced from sucrose hydrolysis (Dtotal).
G. Assay the sample prepared in C (with invertase). The
value reported is the sum of the free glucose and that I. Calibrate frequently as described in the Operations
produced from sucrose hydrolysis (Dtotal). Manual.

H. Calibrate frequently as described in the Operations III. CALCULATIONS

Manual. Case #1
To calculate % glucose, multiply the reported value (Dfree)
Case #2 by the appropriate dilution factor.
The product is a hard candy that is believed to contain 13%
sucrose and 12% glucose. With this sample two separate Example: A 10.10 g powdered seasoning mix sample was
dilutions are necessary. This is due to the glucose membrane prepared as described in II. B and C. When assayed, the
reading both free glucose and the glucose produced from value reported (Dfree) was 14.6 g/L glucose.
the hydrolysis of sucrose. The sum of these two concentra-
tions exceeds the linear range. When analyzing the sample
% Glucose: = 0.1446 gram glucose/gram
treated with invertase a more dilute sample will be needed. powdered mix
14.6 g/L x 0.100
The following sample preparation was used:
L/10.10 g = 14.5% (w/w)
A. Grind the sample into a fine powder.

B. Transfer 10 grams (record exact weight) of sample,

To calculate % sucrose, subtract Dfree from Dtotal and multiply
from step A into a 100 mL volumetric flask using buffer
by the appropriate dilution and mass ratio factors.
diluent to rinse and dilute. Fill the flask to the mark
with buffer and mix. Allow the solution to equilibrate
When the sample containing invertase was assayed, the
for about 20 minutes.
value reported was 15.8 g/L (Dtotal) glucose.

C. Transfer 5 grams (record exact weight) of sample,

from step A into a 100 mL volumetric flask using buffer % Sucrose:
diluent to rinse and dilute. Fill the flask to the mark with (15.8 g/L - 14.6 g/L) x 0.100 L x 342.30 g/L sucrose
buffer and mix. 10.10g 180.16g/L glucose

D. Remove about 3 mL of the solution from C and add = 0.0226 gram sucrose/gram powdered mix
~2 mg of invertase enzyme. Stir gently until dissolved. = 2.26% (w/w)
Cover the sample and allow to incubate at room
temperature for about 20 minutes.

E. Calibrate the 2900 Series with a 1.80 g/L glucose

standard solution.

13 YSI Life Sciences | Application Note 204LS

 Application Note 204LS

Case #2
To calculate % glucose, multiply the reported value (Dfree)
by the appropriate dilution factor.

Example: A 10.10 g ground hard candy sample was

prepared as described in II B. When assayed the value
reported (Dfree) was 12.3 g/L glucose.

% Glucose:
(12.3 g/L x 0.100 L /10.10 g = 0.1218 gram glucose/
gram candy
= 12.2% (w/w)

To calculate % sucrose, multiply the reported value (Dtotal)

by the appropriate dilution factor. Then subtract Dfree from
Dtotal and multiply by the mass ratio factor.

Example: A 5.05 g ground hard candy sample was

prepared as described in II D. When assayed the value
reported (Dtotal) was 13.5 g/L.

% Sucrose:
(13.5 g/L x 0.100 L/5.05 g = 0.2673 gram glucose/
gram candy ORDERING INFORMATION
= 26.7% YSI Part Numbers:
26.7% - 12.2% = 14.5% 2900 Biochemistry Analyzer
2365 Glucose Membrane Kit
342.30 g/mole sucrose 2747 Glucose Standard Solution (1.80g/L)
14.5% x 1531 Glucose Standard Solution (9.0g/L)
180.16g/mole glucose
2357 Buffer Kit
= 27.5% (w/w) sucrose 2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

14 YSI Life Sciences | Application Note 204LS

 Application Note 205LS

Simultaneous Measurement of Glucose and

Sucrose in Peanut Butter
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
complex matrices such as peanut butter can be measured in reagent water).
directly and quickly using the YSI 2900 Series Biochemistry
Analyzer. YSI’s unique enzyme technology provides for rapid D. Connect the 2900 Series instrument to a suitable
glucose and sucrose measurements. Measurements are power source.
virtually unaffected by color, turbidity, density, pH, or the
presence of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a glucose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because glucose
interferes with sucrose analysis, it is necessary to follow G. The following instrument setup is recommended:
this protocol when analyzing for sucrose in the presence
of glucose. Sample size: 10 μL

When a sample is injected into the sample chamber, the Probe A Parameters
sucrose diffuses to the sucrose membrane, which contains Chemistry Glucose
invertase, mutarotase, and glucose oxidase. The sucrose is Unit g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase Calibrator 2.50 g/L
allows for the quick equilibrium of glucose between its α End Point 30 Sec
and β forms. In the presence of glucose oxidase, the β-D-
glucose (glucose) is oxidized to hydrogen peroxide and Probe B Parameters
D-glucono-δ-lactone. The hydrogen peroxide is detected
Chemistry Glucose
amperometrically at the platinum electrode surface. The
Unit g/L
glucose in the sample diffuses to both the glucose and
Calibrator 5.00 g/L
sucrose membranes, which contain glucose oxidase, and
End Point 30 Sec
is oxidized as well. Subtracting the response of the glucose
membrane from the response of the sucrose membrane
yields the response due to sucrose alone. The glucose Autocal Parameters
response is taken directly from the glucose membrane. Temperature 1°C
The algorithm in the instrument software calculates the net Time 30 Min
concentrations. For more information on this system, refer Sample 5 Sam
to the Operations Manual. Cal Shift 2%

I. MATERIALS & SETUP

II. METHOD
A. YSI 2900 Series Biochemistry Analyzer - equipped
A. Weigh between 5.00 and 10.0 grams of peanut
with a 2703 Sucrose Membrane, a 2365 Glucose
butter to be analyzed
Membrane and 2357 Buffer.

B. Transfer the sample to a beaker and add about 50

B. Glucose (2.50 g/L, 25.00 g/L) and Sucrose (5.00 g/L,
mL of buffer. Allow the sample to stir until the peanut
25.0 g/L) standard solutions.
butter is in solution

15 YSI Life Sciences | Application Note 205LS

 Application Note 205LS

C. Transfer the sample to a 100 mL volumetric flask Example: 6.65 g of peanut butter was diluted to 100 mL in a
using buffer diluent to rinse and dilute. Fill the flask Class A volumetric flask. When assayed, the values reported
to the mark with buffer and mix. Allow the solution to were 1.21 g/L glucose and 3.22 g/L sucrose.
equilibrate for about twenty minutes before analysis.
% Glucose: = 0.0182 g glucose/g peanut
D. Calibrate the 2900 Series instrument with 2.50 g/L butter
1.21 g/L x 0.100
glucose and 5.00 g/L sucrose standard solutions.
L/6.65 g = 1.82% (w/w)
E. Check the linearity of the membrane at least once a
day by injection of glucose (25.0 g/L) and sucrose (25.0
% Sucrose: = 0.0484 g sucrose/g peanut
g/L) linearity check solutions. Refer to the Operations
3.22 g/L x 0.100 butter
Manual for specifications.
L/6.65 g = 4.84% (w/w)
F. Assay the sample prepared in C by aspiration into
the 2900 Series instrument.*

G. Calibrate frequently as described in the Operations

Manual. ORDERING INFORMATION
YSI Part Numbers:
* The linear range of the system is 0 to 25.0 g/L for both glucose and 2900 Biochemistry Analyzer
sucrose. The combined concentration of glucose + sucrose cannot 2365 Glucose Membrane Kit
exceed 25 g/L. If the sum of the values reported exceeds this, further
2776 Glucose Standard Solution (2.50g/L)
dilution of the sample is required. If the glucose concentration exceeds
the sucrose concentration, accuracy and precision may be compromised
2777 Glucose Standard Solution (25.0g/L)
due to the software algorithm that subtracts glucose from sucrose. To avoid 2703 Sucrose Membrane Kit
compromising accuracy, refer to Application Note 204LS. 2780 Sucrose Standard Solution (5.00g/L)
2778 Sucrose Standard Solution (25.0g/L)
III. CALCULATIONS 2357 Buffer Kit
To calculate % glucose and sucrose, multiply the values 2363 Potassium Ferrocyanide Test Solution
reported by the appropriate dilution factor. 2392 NaCl Solution (for membrane installation) n

16 YSI Life Sciences | Application Note 205LS

 Application Note 206LS

Lactose Measurement III. METHOD

in Cheese A. Determine an appropriate dilution factor. The sample
injected should contain 1 to 10 g/L lactose.
INTRODUCTION
Lactose concentrations in complex matrices such as cheese B. Accurately weigh the cheese sample to be analyzed.
can be measured directly and quickly using the YSI Model Tr a n s f e r t h e s a m p l e t o a b e a ke r a n d a d d
2900 Series Biochemistry Analyzer. Measurements are approximately 50 mL deionized water.
virtually unaffected by color, turbidity, density, pH, or the
presence of reducing substances. Stir the sample over low heat (~55°C) for 10 to 15 minutes.
Transfer the sample to a 100 mL volumetric flask,
When a sample is injected into the sample chamber, the using deionized water to aid in the complete transfer.
lactose diffuses into the membrane containing galactose Fill the flask to the mark with deionized water and invert
oxidase. The lactose is immediately oxidized to hydrogen to mix.
peroxide and a galactose dialdehyde derivative. The
hydrogen peroxide is detected amperometrically at C. Calibrate the 2900 Series instrument with a 5.00 g/L
the platinum electrode surface. The current flow at the lactose standard solution.
electrode is directly proportional to the hydrogen peroxide
concentration, and hence to lactose concentration. D. Check the linearity of the membrane at least once a
day by injection of a lactose linearity check solution (25.0
I. MATERIALS & SETUP g/L). Refer to the Operations Manual for specifications.
A. YSI Model 2900 Series Biochemistry Analyzer
equipped with a 2702 Galactose Oxidase Membrane E. Assay the sample prepared in B by aspiration into
and 2705 Buffer. the 2900 series instrument. The linear range of the
system is 0.05 to 25.0 g/L lactose. If the value reported
B. Lactose standards (5.00 g/L, 25.0 g/L). exceeds this, further dilution is required.

C. Connect the 2900 Series instrument to a suitable F. Calibration should be done frequently as described
power source. in the Operations Manual.

D. Perform the instrument and membrane daily checks IV. CALCULATIONS

described in the Operations Manual. To calculate % lactose, multiply the reported result by the
appropriate dilution factor.
E. Volumetric glassware (Class A recommended).
Example: 9.96 g of cheese was prepared as described.
F. The following instrument setup is recommended: When assayed, the value reported was 2.16 g/L lactose.
Sample size: 25 μL
% Lactose: = 0.0217 g lactose/cheese
Probe A Parameters Autocal Parameters 2.16 g/L x 0.100
L/9.96 g = 2.17% (w/w)
Chemistry Lactose Temperature 1°C
Unit g/L Time 30 Min
Calibrator 5.00 g/L Sample 5 Sam ORDERING INFORMATION
End Point 30 Sec Cal Shift 2% YSI Part Numbers:
2900 Biochemistry Analyzer
II. LIMITATIONS 2935 Opaque Buffer Bottle
Because galactose and other galactosides (raffinose, 2702 Galactose Oxidase Membrane Kit
stachyose) are also substrates of galactose oxidase, the 2783 Lactose Standard Solution (5.00 g/L)
presence of these in the sample will interfere with this analysis. 2784 Lactose Standard Solution (25.0 g/L)
2705 Buffer Kit
2392 NaCl Solution (for membrane installation) n
17 YSI Life Sciences | Application Note 206LS
 Application Note 207LS

L–Glutamate Determination
INTRODUCTION II. METHOD
L-Glutamate concentrations in complex matrices can A. Total glutamate concentration should not exceed
be measured directly and quickly using the YSI 2900 10.0 mmol/L, as determined in Part D below; otherwise
Series Biochemistry Analyzer. YSI’s unique enzyme the sample will require further dilution. Use volumetric
technology provides for specific L-glutamate measurement. glassware for all dilutions.
Measurements are virtually unaffected by color, turbidity,
density, pH, or the presence of reducing substances. B. Calibrate the 2900 Series instrument with a 5.00
mmol/L L-Glutamate Calibration Standard.
When a sample is injected into the sample chamber, the
glutamate diffuses into the membrane containing glutamate C. Check the linearity of the membrane at least once
oxidase. The glutamate is immediately oxidized to hydrogen a day by injection of a glutamate linearity check
peroxide, α-ketoglutarate, and ammonia. The hydrogen solution (10.0 mmol/L). Refer to the Operators
peroxide is detected amperometrically at the platinum Manual for specifications.
electrode surface. The current flow at the electrode is directly
proportional to the hydrogen peroxide concentration, and D. Assay the sample by aspiration into the 2900 Series.
hence to glutamate concentration. The linear range of the system is 0.1 to 10.0 mmol/L
glutamate. If the value reported exceeds this, further
I. MATERIALS & SETUP dilution is required.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2754 Glutamate Membrane and 2357 Buffer. E. Calibrate frequently as described in the Operations
Manual.
B. L-Glutamate standards (5.00 mmol/L, 10.0 mmol/L).
III. CALCULATIONS
C. Connect the 2900 Series instrument to a suitable To calculate % glutamate, multiply the reported value by the
power source. appropriate dilution factor.

D. Perform the instrument and membrane daily checks For the examples, glutamate concentrations are expressed
described in the Operations Manual. as monosodium glutamate. The molecular weight of
monosodium glutamate (MSG) is 187.13 g/mole or 0.18713
E. Volumetric glassware (Class A recommended). g/mmol.

F. The following instrument setup is recommended. Example: The contents of a can of soup were blended in a
Sample size:25 μL blender on medium speed for about 3 minutes. 10.09 g of
blended soup was diluted to 100 mL in a Class A volumetric
Probe A Parameters Autocal Parameters flask with water. When assayed, the value reported was 1.42
Chemistry Glutamate Temperature 1°C mmol/L glutamate.
Unit mmo/L Time 30 Min
Calibrator 5.00 mmo/L Sample 2 Sam
End Point 30 Sec Cal Shift 2%

18 YSI Life Sciences | Application Note 207LS

 Application Note 207LS

ORDERING INFORMATION
% Glutamate:
= 0.0027 g MSG/g soup YSI Part Numbers:
1.42 mmo/L x
2900 Biochemistry Analyzer
0.100L/10.09g x
2754 Glutamate Oxidase Membrane Kit
0.18713g/mmol = 0.27% (w/w)
2755 L-Glutamate Standard Solution (5.00 mmol/ L)
MSG
2756 L-Glutamate Standard Solution (10.0 mmol/ L)
2357 Buffer Kit
Example: 2.50 grams of a dry powder seasoning mix was 2363 Potassium Ferrocyanide Test Solution
diluted to 100 mL in a Class A volumetric flask with water. 2392 NaCl Solution (for membrane installation) n
The mixture was stirred for 5 minutes. When assayed, the
value reported was 6.85 mmol/L glutamate.

6.85 mmo/L x = 0.0512 g MSG/g seasoning mix

0.100L/2.50g x
0.18713g/mmol
MSG = 5.12% (w/w)

19 YSI Life Sciences | Application Note 207LS

 Application Note 208LS

Determination of Hydrogen Probe A Parameters Autocal Parameters

Chemistry Peroxide Temperature 1°C
Peroxide Unit mg/L (ppm) Time 30 Min
Calibrator ~30 (see section B left) Sample 5 Sam
INTRODUCTION End Point 30 Sec Cal Shift 2%
Hydrogen peroxide can be measured quickly using the YSI
2900 Series Biochemistry Analyzer. YSI’s unique technology * The sample volume can be changed to meet the specific needs. Low
hydrogen peroxide concentrations will require larger sample volumes.
provides for rapid hydrogen peroxide determination.
Measurements are virtually unaffected by color, turbidity, The calibration solution should be stored and sampled from glass
density, or pH. containers. However, if the solution is prepared and used the
same day, calibrate from Station #2 and sample from test tubes.

When a sample is injected into the sample chamber, the

II. METHOD
hydrogen peroxide diffuses to the platinum electrode and
A. Dilute samples with distilled water to bring the
is oxidized. The current flow at the electrode is directly
hydrogen peroxide concentration below 300 ppm.
proportional to hydrogen peroxide concentration. The
blank membrane placed over the electrode surface rejects
B. Calibrate the 2900 Series instrument with the
potential interfering substances.
hydrogen peroxide calibration standard prepared
in I.B.
Low molecular weight phenols, mercaptans, hydroxylamines,
hydrazines, and anilines can be electrochemical interferences.
C. If desired, check the linearity of the membrane by
Refer to the Operations Manual for specifics.
injection of the linearity solution prepared in II.B.
Typically, hydrogen peroxide response is linear from
I. MATERIALS & SETUP
3 to 300 ppm.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2701 Blank Membrane and 2357 Buffer.
D. Assay the sample prepared in II.A. by aspiration into
the 2900 Series. If the value reported exceeds 300 ppm,
B. Hydrogen Peroxide Standards. YSI does not offer
dilute the sample further.
hydrogen peroxide standards. Prepare a calibration
standard with a hydrogen peroxide concentration near
E. Calibrate frequently as described in the Operations
the expected assay value of the sample, however, the
Manual.
calibration standard must produce more than 5 nA of
current. In general, about 15 ppm (mg/L) hydrogen
III. CALCULATIONS
peroxide is the lower limit for a calibration standard.
To calculate % hydrogen peroxide, multiply the reported
value by the appropriate dilution factor.
A linearity standard can also be prepared. Target
a concentration that reflects the highest hydrogen Example: 10.0 ml of sample was diluted to 100 mL in a Class
peroxide concentration of the samples being analyzed. A volumetric flask. When assayed, the value reported was
220 mg/L (ppm).
C. Connect the 2900 Series instrument to a suitable
power source. Hydrogen Peroxide in original sample:
220 mg/L x 0.100L/0.010L = 2200 mg/L (ppm)
D. Perform the instrument and membrane check
described in the Operations Manual.
ORDERING INFORMATION
E. Volumetric glassware (Class A recommended). YSI Part Numbers:
2900 Biochemistry Analyzer
F. The following instrument setup is recommended. 2701 Blank Membrane Kit
Sample size 25 μL* 2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

20 YSI Life Sciences | Application Note 208LS

 Application Note 209LS

Simultaneous Measurement of Glucose

and Sucrose in Frozen Ice Cream Bars
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
complex matrices such as ice cream bars can be measured in reagent water).
directly and quickly using the YSI 2900 Series Biochemistry
Analyzer. YSI’s unique enzyme technology provides for rapid D. Connect the 2900 Series instrument to a suitable power
glucose and sucrose measurements. Measurements are source.
virtually unaffected by color, turbidity, density, pH, or the
presence of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a glucose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because glucose
interferes with sucrose analysis, it is necessary to follow G. The following instrument setup is recommended:
this protocol when analyzing for sucrose in the presence Sample Size 10 μL
of glucose.
Probe A Parameters Probe B Parameters
When a sample is injected into the sample chamber, the Chemistry Glucose Chemistry Sucrose
sucrose diffuses to the sucrose membrane, which contains Unit g/L Unit g/L
invertase, mutarotase, and glucose oxidase. The sucrose is Calibrator 2.50 Calibrator 5.00 g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase End Point 30 Sec End Point 30 Sec
allows for the quick equilibrium of glucose between its α
and β forms. In the presence of glucose oxidase, the β-D- Autocal Parameters
glucose (glucose) is oxidized to hydrogen peroxide and Temperature 1°C
D-glucono-δ-lactone. The hydrogen peroxide is detected Time 30 Min
amperometrically at the platinum electrode surface. The Sample 5 Sam
glucose in the sample diffuses to both the glucose and Cal Shift 2%
sucrose membranes, which contain glucose oxidase, and
is oxidized as well. Subtracting the response of the glucose
membrane from the response of the sucrose membrane II. METHOD
yields the response due to sucrose alone. The glucose A. Separate the chocolate coating from the ice cream.
response is taken directly from the glucose membrane.
The algorithm in the instrument software calculates the net B. Transfer the chocolate coating to a container that is water
concentrations. For more information on this system, refer resistant and can be closed, such as a bottle or a plastic
to the Operations Manual. bag.

I. MATERIALS & SETUP C. Immerse the container into a beaker of hot water
A. YSI 2900 Series Biochemistry Analyzer - equipped (55-65°C) making sure no water enters the container.
with a 2703 Sucrose Membrane, a 2365 Glucose Allow the sample to melt stirring occasionally to keep
Membrane and 2357 Buffer. the sample homogeneous.

B. Glucose (2.50 g/L, 25.00 g/L) and Sucrose (5.00 g/L, D. Transfer about 5 grams (record exact weight) of the
25.0 g/L) standard solutions. chocolate coating to a 100 mL volumetric flask, using
buffer diluent to rinse and dilute. Fill the flask to
the mark with buffer and mix. Allow the solution to
equilibrate for about 20 minutes.
21 YSI Life Sciences | Application Note 209LS
 Application Note 209LS

E. Transfer the ice cream (without coating) into a water

resistant container and immerse in hot water until the % Glucose: = 0.0054 gram glucose/gram
0.545 g/L x 0.100L ice cream
sample is melted. Stir occasionally to keep the sample
homogeneous. /10.10 g = 0.54% (w/w)

F. Transfer about 10 grams (record exact weight) of the ice

cream sample to a 100 mL volumetric flask using buffer % Sucrose: = 0.0983 gram glucose/gram
9.93 g/L x 0.100L ice cream
diluent to rinse and dilute. Fill the flask to the mark with
buffer and mix. Allow the solution to equilibrate for /10.10 g = 9.83% (w/w)
about 20 minutes.
Example: 5.01 grams of chocolate coating were diluted to
G. Calibrate the 2900 Series instrument with 2.50 100 mL in a class A volumetric flask. When assayed, the values
g/L glucose and 5.00 g/L sucrose standard solutions. reported were 0.118 g/L glucose and 8.73 g/L sucrose.

H. Check the linearity of the membrane at least = 0.0024 gram glucose/gram

% Glucose:
once a day by injection of glucose (25.00 g/L) and coating
0.118 g/L x 0.100L
sucrose (25.0 g/L) linearity check solutions. Refer to the
/5.01 g = 0.24% (w/w)
Operations Manual for specifications.

I. Shake or stir the sample prepared in D, to keep the

% Sucrose: = 0.1743 gram glucose/gram
sample homogeneous. Assay the sample prepared in coating
8.73 g/L x 0.100L
D, by aspiration into the 2900 Series instrument. Then
/5.01 g = 17.4% (w/w)
assay the sample prepared in F by aspiration into the
2900 Series.*

J. Calibrate frequently as described in the Operations ORDERING INFORMATION

Manual. YSI Part Numbers:
2900 Biochemistry Analyzer
* The linear range of the system is 0 to 25.0 g/L for both glucose and
2365 Glucose Membrane Kit
sucrose. The combined concentration of glucose + sucrose cannot
exceed 25 g/L. If the sum of the values reported exceeds this, further
2776 Glucose Standard Solution (2.50 g/L)
dilution of the sample is required. If the glucose concentration exceeds 2777 Glucose Standard Solution (25.00 g/L)
the sucrose concentration accuracy and precision may be compromised 2703 Sucrose Membrane Kit
due to the software algorithm that subtracts glucose from sucrose. To avoid 2780 Sucrose Standard Solution (5.00 g/L)
compromising accuracy refer to Application Note 204LS.
2778 Sucrose Standard Solution (25.0 g/L)
2357 Buffer Kit
III. CALCULATIONS 2363 Potassium Ferrocyanide Test Solution
To calculate % glucose and sucrose, multiply the values 2392 NaCl Solution (for membrane installation) n
reported by the appropriate dilution factor.

Example: 10.10 grams of ice cream were diluted to 100

mL in a Class A volumetric flask. When assayed, the values
reported were 0.545 g/L glucose and 9.93 g/L sucrose.

22 YSI Life Sciences | Application Note 209LS

 Application Note 210LS

Glucose Measurement in Canned Green Beans

INTRODUCTION II. METHOD
Dextrose (D-glucose) concentrations in complex matrices A. Empty contents of can into a clean dry blender.
such as canned green beans can be measured directly and
quickly using the YSI 2900 Series Biochemistry Analyzer. YSI’s B. Blend until the sample is liquid, about ten minutes.
unique enzyme technology provides for specific glucose
measurement. Measurements are virtually unaffected by C. Transfer 25 to 50 grams of the blended sample to a
color, turbidity, density, pH, or the presence of reducing 100 mL volumetric flask, using buffer diluent to rinse
substances. and dilute. Fill the flask to the mark with buffer and
mix. Allow the solution to equilibrate for about twenty
When a sample is injected into the sample chamber, the minutes before analysis.
glucose diffuses into the membrane containing glucose
oxidase. The glucose is immediately oxidized to hydrogen D. Calibrate the 2900 Series instrument with a 2.50 g/L
peroxide and D-glucono-δ-lactone. The hydrogen peroxide glucose standard solution.
is detected amperometrically at the platinum electrode
surface. The current flow at the electrode is directly E. Check the linearity of the membrane at least once a
proportional to the hydrogen peroxide concentration, and day by injection of a glucose linearity check solution (9.00
hence to glucose concentration. g/L). Refer to the Operators Manual for specifications.

I. MATERIALS & SETUP F. Assay the sample prepared in C by aspiration into the
A. YSI 2900 Series Biochemistry Analyzer - equipped 2900 Series. The linear range of the system is 0.05 to
with a 2365 Glucose Membrane and 2357 Buffer. 9.00 g/L glucose. If the value reported exceeds this,
further dilution is required.*
B. Glucose standards (2.50 g/L, 9.00 g/L).
G. Calibrate frequently as described in the Operations
C. Buffer Diluent (40 g/L NaH 2PO 4, 10 g/L Na 2HPO 4 Manual.
in reagent water).
* The linearity of glucose on the 2900 series instrument can be increased
D. Connect the 2900 Series instrument to a suitable power to 0.05 to 25.0 g/L. This can be done by decreasing the sample size to 10
μL and checking the linearity with a 25.0 g/L solution.
source.

E. Perform the instrument and membrane daily checks III. CALCULATIONS

described in the Operations Manual. To calculate % glucose, multiply the reported value by the
appropriate dilution factor.
F. Volumetric glassware (Class A recommended).
Example: 50.02 grams of blended green beans were dilut-
G. The following instrument setup is recommended: ed to 100 mL in a Class A volumetric flask. When assayed,
Sample Size 25 μL the value reported was 2.93 g/L glucose.

Probe A Parameters Autocal Parameters

% Glucose: = 0.0059 gram glucose/gram
Chemistry Glucose Temperature 1°C green beans
2.93 g/L x 0.100L
Unit g/L Time 30 Min
/50.02 g = 0.59% (w/w)
Calibrator 2.50 g/L Sample 5 Sam
End Point 30 Sec Cal Shift 2%

23 YSI Life Sciences | Application Note 210LS

 Application Note 210LS

ORDERING INFORMATION
YSI Part Numbers:
2900 Biochemistry Analyzer
2365 Glucose Membrane Kit
2776 Glucose Standard Solution (2.50 g/L)
1531 Glucose Standard Solution (9.00 g/L)
2777 Glucose Standard Solution (25.0 g/L)
2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

24 YSI Life Sciences | Application Note 210LS

 Application Note 211LS

Glucose Measurement in Frozen Green Beans

INTRODUCTION
Dextrose (D-glucose) concentrations in complex matrices Autocal Parameters
such as frozen green beans can be measured directly and Temperature 1°C
quickly using the YSI 2900 Series Biochemistry Analyzer. YSI’s Time 30 Min
unique enzyme technology provides for specific glucose Sample 5 Sam
measurement. Measurements are virtually unaffected by Cal Shift 2%
color, turbidity, density, pH, or the presence of reducing
substances.
II. METHOD
When a sample is injected into the sample chamber, the A. Weigh up to 25.00 g of green beans to be analyzed,
glucose diffuses into the membrane containing glucose add 50 mL of buffer.
oxidase. The glucose is immediately oxidized to hydrogen
peroxide and D-glucono-δ-lactone. The hydrogen peroxide B. Transfer the sample a clean dry blender. Blend until
is detected amperometrically at the platinum electrode the sample is liquid.
surface. The current flow at the electrode is directly
proportional to the hydrogen peroxide concentration, and C. Transfer the sample to a 100mL volumetric flask using
hence to glucose concentration. buffer diluent to rinse and dilute. Fill the flask to the
mark with buffer. Allow the solution to equilibrate for
I. MATERIALS & SETUP about twenty minutes before analysis.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2365 Glucose Membrane and 2357 Buffer. D. Calibrate the 2900 Series instrument with a 2.50 g/L
glucose standard solution.
B. Glucose standards (2.50 g/L, 9.00 g/L).
E. Check the linearity of the membrane at least once a day
C. Buffer Diluent (40 g/L NaH 2PO 4, 10 g/L Na 2HPO 4 by injection of a glucose linearity check solution (9.00
in reagent water). g/L). Refer to the Operators Manual for specifications.

D. Connect the 2900 Series instrument to a suitable power F. Assay the sample prepared in C by aspiration into the
source. 2900 Series. The linear range of the system is 0.05 to
9.00 g/L glucose. If the value reported exceeds this,
E. Perform the instrument and membrane daily checks further dilution is required.*
described in the Operations Manual.
G. Calibrate frequently as described in the Operations
F. Volumetric glassware (Class A recommended). Manual.

G. The following instrument setup is recommended: * The linearity of glucose on the 2900 Series can be increased to 0.05 to
25.0 g/L. This can be done by decreasing the sample size to 10μL and
Sample Size 25 μL
checking the linearity with 25.0 g/L standard.

Probe A Parameters
III. CALCULATIONS
Chemistry Glucose To calculate % glucose, multiply the reported value by the
Unit g/L appropriate dilution factor.
Calibrator 2.50 g/L
End Point 30 Sec

25 YSI Life Sciences | Application Note 211LS

 Application Note 211LS

Example: 27.22 grams of frozen green beans were diluted ORDERING INFORMATION
to 100 mL in a Class A volumetric flask. When assayed, the YSI Part Numbers:
value reported was 2.20 g/L glucose. 2900 Biochemistry Analyzer
2365 Glucose Membrane Kit
2776 Glucose Standard Solution (2.50 g/L)
% Glucose: = 0.0081 g glucose/ 1531 Glucose Standard Solution (9.00 g/L)
2.20 g/L x 0.100L g green beans 2777 Glucose Standard Solution (25.0 g/L)
/27.22 g = 0.81% (w/w) 2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

26 YSI Life Sciences | Application Note 211LS

 Application Note 212LS

L-Lactate in Lunch Meats

INTRODUCTION
L-Lactate concentrations in complex matrices such as lunch Autocal Parameters
meats can be measured directly and quickly using the YSI Temperature 1°C
2900 Series Biochemistry Analyzer. YSI’s unique enzyme Time 30 Min
technology provides for specific L-lactate measurement. Sample 5 Sam
Measurements are virtually unaffected by color, turbidity, Cal Shift 2%
density, pH, or the presence of reducing substances.

When a sample is injected into the sample chamber, the II. METHOD
L-lactate diffuses into the membrane containing L-lactate A. Cut the sample into several pieces (about 1 inch squares).
oxidase. The L-lactate is immediately oxidized to hydrogen
peroxide and pyruvate. The hydrogen peroxide is detected B. Weigh the cut pieces of meat and record the exact weight.
amperometrically at the platinum electrode surface. The
current flow at the electrode is directly proportional to the C. Transfer the sample to a clean dry blender. Add about
hydrogen peroxide concentration, and hence to the L-lactate 100 mL of distilled or deionized water. Turn the blender
concentration. on and allow the sample to blend for about 5 minutes.

I. MATERIALS & SETUP D. Transfer the sample to a 500 mL volumetric flask. Using
A. YSI 2900 Series Biochemistry Analyzer - equipped distilled or deionized water, rinse the blender and use
with a 2329 L-Lactate Membrane and 2357 Buffer. this rinse to dilute the sample to the mark on the flask.

B. L-Lactate standards (0.50 g/L, 2.50 g/L). E. Calibrate the 2900 Series instrument with a 0.50
g/L L-lactate standard solution.
C. Connect the 2900 Series instrument to a suitable power
source. F. Assay the sample prepared in D by aspiration into
the 2900 Series. The linear range of the system extends
D. Perform the instrument and membrane daily checks to 2.50 g/L L-lactate. If the value reported exceeds this,
described in the Operations Manual. further dilution is required.

E. Volumetric glassware (Class A recommended). G. Check the linearity of the membrane at least once a day
by injection of an L-lactate linearity check solution (2.50
F. The following instrument setup is recommended: g/L). Refer to the Operations Manual for specifications.
Sample Size 25 μL
H. Calibrate frequently as described in the Operations
Probe A Parameters Manual.
Chemistry L-Lactate
Unit g/L
Calibrator 0.50 g/L
End Point 30 Sec

27 YSI Life Sciences | Application Note 212LS

 Application Note 212LS

III. CALCULATIONS
27.93 grams of turkey lunch meat was diluted to 500 mL in
a Class A volumetric flask. When assayed the value reported
was 1.27 g/L L-lactate.

% L-Lactate = 0.0227 g L-Lactate/g Meat

1.27 g/L x 0.500L
/27.93 g = 2.27% (w/w)

To measure sodium lactate multiply the concentration by the

ratio of the formula weights of sodium lactate and L-lactate. ORDERING INFORMATION
YSI Part Numbers:
2900 Biochemistry Analyzer
Sodium lactate = 112.07 g/mole
2329 L-Lactate Membrane Kit
L-lactate = 89.07 g/mole 2776 L-Lactate Standard Solution (0.50 g/L)
112.07 / 89.07 = 1.26 2777 L-Lactate Standard Solution (2.50 g/L)
2.27%(w/w) L-lactate = 2.86% (w/w) 2357 Buffer Kit
x 1.26 sodium lactate 2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

28 YSI Life Sciences | Application Note 212LS

 Application Note 213LS

Simultaneous Measurement of Glucose

and Sucrose in Corn and Peas
INTRODUCTION
Glucose (D-glucose) and sucrose concentrations in complex C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
matrices such as corn and peas can be measured directly in reagent water).
and quickly using the YSI 2900 Series Biochemistry Analyzer.
YSI’s unique enzyme technology provides for rapid glucose D. Connect the 2900 Series instrument to a suitable power
and sucrose measurements. Measurements are virtually source.
unaffected by color, turbidity, density, pH, or the presence
of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a glucose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because glucose
interferes with sucrose analysis, it is necessary to follow G. The following instrument setup is recommended:
this protocol when analyzing for sucrose in the presence Sample Size 10 μL
of glucose.
Probe A Parameters Probe B Parameters
When a sample is injected into the sample chamber, the Chemistry Glucose Chemistry Sucrose
sucrose diffuses to the sucrose membrane, which contains Unit g/L Unit g/L
invertase, mutarotase, and glucose oxidase. The sucrose is Calibrator 2.50 Calibrator 5.00 g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase End Point 30 Sec End Point 30 Sec
allows for the quick equilibrium of glucose between its α
and β forms. In the presence of glucose oxidase, the β-D- Autocal Parameters
glucose (glucose) is oxidized to hydrogen peroxide and Temperature 1°C
D-glucono-δ-lactone. The hydrogen peroxide is detected Time 30 Min
amperometrically at the platinum electrode surface. The Sample 5 Sam
glucose in the sample diffuses to both the glucose and Cal Shift 2%
sucrose membranes, which contain glucose oxidase, and
is oxidized as well. Subtracting the response of the glucose II. METHOD
membrane from the response of the sucrose membrane A. Weigh up to 25.00 grams of corn or peas to be
yields the response due to sucrose alone. The glucose analyzed. Add about 50 mL buffer diluent.
response is taken directly from the glucose membrane.
The algorithm in the instrument software calculates the net B. Transfer the solution to a clean dry blender. Blend
concentrations. For more information on this system, refer until the sample is liquid.
to the Operations Manual.
C. Transfer the sample to a 100 mL volumetric flask
I. MATERIALS & SETUP using buffer diluent to rinse and dilute. Fill the flask
A. YSI 2900 Series Biochemistry Analyzer - equipped to the mark with buffer and mix. Allow the solution to
with a 2703 Sucrose Membrane, a 2365 Glucose equilibrate for about twenty minutes before analysis.
Membrane and 2357 Buffer.
D. Calibrate the 2900 Series instrument with a 2.50 g/L
B. Glucose (2.50 g/L, 25.00 g/L) and Sucrose (5.00 g/L, glucose and 5.00 g/L sucrose standard solutions.
25.0 g/L) standard solutions.

29 YSI Life Sciences | Application Note 213LS

 Application Note 213LS

E. Check the linearity of the membranes at least once

a day by injection of glucose (25.00 g/L) and sucrose % Glucose: = 0.0005 g glucose/g peas
(25.0 g/L) linearity check solutions. Refer to the 0.128 g/L x 0.100L
/25.10 g = 0.05% (w/w)
Operations Manual for specifications.

F. Assay the sample prepared in C by aspiration into the

2900 Series instrument.*
% Sucrose: = 0.0275 g sucrose/g peas
G. Calibrate frequently as described in the Operations 6.91 g/L x 0.100L
/25.10 g = 2.75% (w/w)
Manual.

* The linear range of the system is 0 to 25.0 g/L for both glucose and
sucrose. The combined concentration of glucose + sucrose cannot exceed
25 g/L. If the sum of the values reported exceeds this, further dilution of
the sample is required.
ORDERING INFORMATION
If the glucose concentration exceeds the sucrose concentration, accuracy
and precision may be compromised due to the software algorithm that
YSI Part Numbers:
subtracts glucose from sucrose. To avoid compromising accuracy refer to 2900 Biochemistry Analyzer
Application Note 204LS. 2365 Glucose Membrane Kit
2776 Glucose Standard Solution (2.50 g/L)
III. CALCULATIONS 2777 Glucose Standard Solution (25.00 g/L)
To calculate % glucose and sucrose, multiply the values 2703 Sucrose Membrane Kit
reported by the appropriate dilution factor. 2780 Sucrose Standard Solution (5.00 g/L)
2778 Sucrose Standard Solution (25.0 g/L)
Example: 25.10 grams of peas were diluted to 100 mL 2357 Buffer Kit
in a Class A volumetric flask. When assayed, the values 2363 Potassium Ferrocyanide Test Solution
reported were 0.128 g/L glucose and 6.91 g/L sucrose. 2392 NaCl Solution (for membrane installation) n

30 YSI Life Sciences | Application Note 213LS

 Application Note 214LS

Simultaneous Measurement of Glucose

and Sucrose in Cereal Products
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
complex matrices such as cereal products can be measured in reagent water).
directly and quickly using the YSI 2900 Series Biochemistry
Analyzer. YSI’s unique enzyme technology provides for D. Connect the 2900 Series instrument to a suitable power
specific glucose and sucrose measurements. Measurements source.
are virtually unaffected by color, turbidity, density, pH, or the
presence of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a glucose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because glucose
interferes with sucrose analysis, it is necessary to follow G. The following instrument setup is recommended:
this protocol when analyzing for sucrose in the presence Sample Size 10 μL
of glucose.
Probe A Parameters Probe B Parameters
When a sample is injected into the sample chamber, the Chemistry Glucose Chemistry Sucrose
sucrose diffuses to the sucrose membrane, which contains Unit g/L Unit g/L
invertase, mutarotase, and glucose oxidase. The sucrose is Calibrator 2.50 Calibrator 5.00 g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase End Point 30 Sec End Point 30 Sec
allows for the quick equilibrium of glucose between its α
and β forms. In the presence of glucose oxidase, the β-D- Autocal Parameters
glucose (glucose) is oxidized to hydrogen peroxide and
Temperature 1°C
D-glucono-δ-lactone. The hydrogen peroxide is detected
Time 30 Min
amperometrically at the platinum electrode surface. The
Sample 5 Sam
glucose in the sample diffuses to both the glucose and
Cal Shift 2%
sucrose membranes, which contain glucose oxidase, and
is oxidized as well. Subtracting the response of the glucose
membrane from the response of the sucrose membrane II. METHOD
yields the response due to sucrose alone. The glucose A. Grind sample to a fine powder.
response is taken directly from the glucose membrane.
The algorithm in the instrument software calculates the net B. Weigh 1.000 to 5.000 g of powdered sample.
concentrations. For more information on this system, refer
to the Operations Manual. C. Transfer the sample to a 100 mL volumetric flask
using buffer diluent to rinse and dilute. Fill the flask
I. MATERIALS & SETUP to the mark with buffer and mix. Allow the solution to
A. YSI 2900 Series Biochemistry Analyzer - equipped equilibrate for about twenty minutes before analysis.
with a 2703 Sucrose Membrane, a 2365 Glucose
Membrane and 2357 Buffer. D. Calibrate the 2900 Series instrument with a 2.50 g/L
glucose and 5.00 g/L sucrose standard solutions.
B. Glucose (2.50 g/L,25.00 g/L) and Sucrose (5.00 g/L,
25.0 g/L) standard solutions. E. Check the linearity of the membranes at least once
a day by injection of glucose (25.00 g/L) and sucrose
(25.0 g/L) linearity check solutions. Refer to the
Operations Manual for specifications.
31 YSI Life Sciences | Application Note 214LS
 Application Note 214LS

F. Assay the sample prepared in B by aspiration into the IV. SAMPLES TESTED
2900 Series instrument.* Several cereal samples were assayed using YSI technology
and HPLC. The results are listed below.
G. Calibrate frequently as described in the Operations
Manual. Sucrose (%) Glucose (%)
Sample YSI HPLC YSI HPLC Label (%)*
* The linear range of the system is 0 to 25.0 g/L for both glucose and
A 35.0 36.0 1.24 0.96 38.8
sucrose. The combined concentration of glucose + sucrose cannot
exceed 25 g/L. If the sum of the values reported exceeds this, further B 21.4 22.2 0.85 0.50 21.1
dilution of the sample is required. If the glucose concentration exceeds C 22.4 23.1 1.52 1.52 24.6
the sucrose concentration, accuracy and precision may be compromised
due to the software algorithm that subtracts glucose from sucrose. To avoid D 17.4 18.3 0.27 0.07 28.0
compromising accuracy refer to Application Note 204LS. E 25.2 25.4 1.61 1.33 28.2
F 29.8 27.9 3.51 4.14 35.2
III. CALCULATIONS
G 32.2 31.1 3.01 2.42 38.7
To calculate % glucose and sucrose, multiply the values
H 32.9 30.4 0.71 0.43 38.7
reported by the appropriate dilution factor.
I 5.40 6.70 1.03 0.70 7.0
Example: A cereal sample (4.336 g) was prepared and * Label information reported as “Sucrose and Other Sugars.”
assayed as described. The values reported were 0.62 g/L
glucose and 9.88 g/L sucrose. ORDERING INFORMATION
YSI Part Numbers:
2900 Biochemistry Analyzer
% Glucose: = 0.0143 g glucose/g cereal 2365 Glucose Membrane Kit
0.62 g/L x 0.100L 2776 Glucose Standard Solution (2.50 g/L)
/4.336 g = 1.43% (w/w)
2777 Glucose Standard Solution (25.00 g/L)
2703 Sucrose Membrane Kit
2780 Sucrose Standard Solution (5.00 g/L)
% Sucrose: 2778 Sucrose Standard Solution (25.0 g/L)
= 0.2279 g sucrose/g cereal
9.88 g/L x 0.100L 2357 Buffer Kit
/4.336 g = 22.8% (w/w) 2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

32 YSI Life Sciences | Application Note 214LS

 Application Note 215LS

Simultaneous Measurement of Glucose

and Sucrose in Baked Goods
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
complex matrices such as baked goods can be measured in reagent water).
directly and quickly using the YSI 2900 Series Biochemistry
Analyzer. YSI’s unique enzyme technology provides for rapid D. Connect the 2900 Series instrument to a suitable power
glucose and sucrose measurements. Measurements are source.
virtually unaffected by color, turbidity, density, pH, or the
presence of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a glucose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because glucose
interferes with sucrose analysis, it is necessary to follow G. The following instrument setup is recommended:
this protocol when analyzing for sucrose in the presence Sample Size 10 μL
of glucose.
Probe A Parameters Probe B Parameters
When a sample is injected into the sample chamber, the Chemistry Glucose Chemistry Sucrose
sucrose diffuses to the sucrose membrane, which contains Unit g/L Unit g/L
invertase, mutarotase, and glucose oxidase. The sucrose is Calibrator 2.50 Calibrator 5.00 g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase End Point 30 Sec End Point 30 Sec
allows for the quick equilibrium of glucose between its α
and β forms. In the presence of glucose oxidase, the β-D- Autocal Parameters
glucose (glucose) is oxidized to hydrogen peroxide and
Temperature 1°C
D-glucono-δ-lactone. The hydrogen peroxide is detected
Time 30 Min
amperometrically at the platinum electrode surface. The
Sample 5 Sam
glucose in the sample diffuses to both the glucose and
Cal Shift 2%
sucrose membranes, which contain glucose oxidase, and
is oxidized as well. Subtracting the response of the glucose
membrane from the response of the sucrose membrane II. METHOD
yields the response due to sucrose alone. The glucose A. Grind sample to a fine powder.
response is taken directly from the glucose membrane.
The algorithm in the instrument software calculates the net B. Weigh 1.00 to 5.00 g of powdered sample.
concentrations. For more information on this system, refer
to the operations manual. C. Transfer the sample to a 100 mL volumetric flask
using buffer diluent to rinse and dilute. Fill the flask
I. MATERIALS & SETUP to the mark with buffer and mix. Allow the solution to
A. YSI 2900 Series Biochemistry Analyzer - equipped equilibrate for about twenty minutes before analysis.
with a 2703 Sucrose Membrane, a 2365 Glucose
Membrane and 2357 Buffer. D. Calibrate the 2900 Series instrument with 2.50 g/L
glucose and 5.00 g/L sucrose standard solutions.
B. Glucose (2.50 g/L, 25.00 g/L) and Sucrose (5.00 g/L,
25.0 g/L) standard solutions.

33 YSI Life Sciences | Application Note 215LS

 Application Note 215LS

E. Check the linearity of the membranes at least once

a day by injection of glucose (25.0 g/L) and sucrose
(25.0 g/L) linearity check solutions. Refer to the
Operations Manual for specifications.

F. Assay the sample prepared in B by aspiration into the

2900 Series instrument.*

G. Calibrate frequently as described in the Operations

Manual.

* The linear range of the system is 0 to 25.0 g/L for both glucose and
sucrose. The combined concentration of glucose + sucrose cannot
exceed 25 g/L. If the sum of the values reported exceeds this, further
dilution of the sample is required. If the glucose concentration exceeds
the sucrose concentration, accuracy and precision may be compromised
due to the software algorithm that subtracts glucose from sucrose. To avoid
compromising accuracy refer to Application Note 204LS.

III. CALCULATIONS
To calculate % glucose and sucrose, multiply the values re-
ported by the appropriate dilution factor.

Example: A baked muffin sample (4.654 g) was prepared

and assayed as described. The values reported were 0.81
g/L glucose and 8.54 g/L sucrose.
ORDERING INFORMATION
YSI Part Numbers:
2900 Biochemistry Analyzer
% Glucose: = 0.0174 g glucose/g muffin 2365 Glucose Membrane Kit
0.81 g/L x 0.100L
= 1.74% (w/w) 2776 Glucose Standard Solution (2.50 g/L)
/4.654 g
2777 Glucose Standard Solution (25.0 g/L)
2703 Sucrose Membrane Kit
2780 Sucrose Standard Solution (5.00 g/L)
% Sucrose: 2778 Sucrose Standard Solution (25.0 g/L)
= 0.1834 g sucrose/g muffin
8.54 g/L x 0.100L 2357 Buffer Kit
/4.654 g = 18.3% (w/w) 2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

34 YSI Life Sciences | Application Note 215LS

 Application Note 216LS

Simultaneous Measurement of Glucose

and Sucrose in Sweetened Condensed Milk
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
complex matrices such as sweetened condensed milk can in reagent water).
be measured directly and quickly using the YSI 2900 Series
Biochemistry Analyzer. YSI’s unique enzyme technology D. Connect the 2900 Series instrument to a suitable
provides for rapid glucose and sucrose measurements. power source.
Measurements are virtually unaffected by color, turbidity,
density, pH, or the presence of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a glucose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because glucose
interferes with sucrose analysis, it is necessary to follow G. The following instrument setup is recommended:
this protocol when analyzing for sucrose in the presence Sample Size 10 μL
of glucose.
Probe A Parameters Probe B Parameters
When a sample is injected into the sample chamber, the Chemistry Glucose Chemistry Sucrose
sucrose diffuses to the sucrose membrane, which contains Unit g/L Unit g/L
invertase, mutarotase, and glucose oxidase. The sucrose is Calibrator 2.50 Calibrator 5.00 g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase End Point 30 Sec End Point 30 Sec
allows for the quick equilibrium of glucose between its α
and β forms. In the presence of glucose oxidase, the β-D- Autocal Parameters
dextrose (glucose) is oxidized to hydrogen peroxide and Temperature 1°C
D-glucono-δ-lactone. The hydrogen peroxide is detected Time 30 Min
amperometrically at the platinum electrode surface. The Sample 5 Sam
glucose in the sample diffuses to both the glucose and Cal Shift 2%
sucrose membranes, which contain glucose oxidase, and
is oxidized as well. Subtracting the response of the glucose
membrane from the response of the sucrose membrane II. METHOD
yields the response due to sucrose alone. The glucose A. As with all applications, make sure the sample is
response is taken directly from the glucose membrane. homogeneous. In the case of a canned sample of
The algorithm in the instrument software calculates the net sweetened condensed milk, warm the can in a water
concentrations. For more information on this system, refer bath that is between 35-45°C. Shake the can before
to the Operations Manual. opening. Transfer the contents of the can to another
container. Scrape the top, bottom and sides of the can,
I. MATERIALS & SETUP making sure to remove any sugar that may have settled.
A. YSI 2900 Series Biochemistry Analyzer - equipped Stir entire contents.
with a 2703 Sucrose Membrane, a 2365 Glucose
Membrane and 2357 Buffer. B. Weigh up to 1.000 g of sweetened condensed milk to
be analyzed.
B. Glucose (2.50 g/L, 25.00 g/L) and Sucrose (5.00 g/L,
25.0 g/L) standard solutions.

35 YSI Life Sciences | Application Note 216LS

 Application Note 216LS

C. Transfer the sample to a 100 mL volumetric flask III. CALCULATIONS

using buffer diluent to rinse and dilute. Fill the flask To calculate % glucose and sucrose, multiply the values re-
to the mark with buffer and mix. Allow the solution to ported by the appropriate dilution factor.
equilibrate for about twenty minutes before analysis.
Example: 1.0135 g of sweetened condensed milk was
D. Calibrate the 2900 Series instrument with 2.50 g/L diluted to 100 mL in a Class A volumetric flask. When as-
glucose and 5.00 g/L sucrose standard solutions. sayed, the values reported were 0.096 g/L glucose and
4.47 g/L sucrose.
E. Check the linearity of the membranes at least once
a day by injection of glucose (25.0 g/L) and sucrose
% Glucose: = 0.0095 g glucose/g milk
(25.0 g/L) linearity check solutions. Refer to the
0.096 g/L x 0.100L
Operations Manual for specifications. = 0.95% (w/w)
/1.0135 g
F. Assay the sample prepared in B by aspiration into the
2900 Series instrument.*
% Sucrose: = 0.4410 g sucrose/g milk
G. Calibrate frequently as described in the Operations 4.47 g/L x 0.100L
/1.0135 g = 44.1% (w/w)
Manual.

* The linear range of the system is 0.05 to 25.0 g/L for both glucose and
sucrose. The combined concentration of glucose + sucrose cannot exceed ORDERING INFORMATION
25.0 g/L. If the sum of the values reported exceeds this, further dilution of
YSI Part Numbers:
the sample is required.
2900 Biochemistry Analyzer
If the glucose concentration exceeds the sucrose concentration, accuracy 2365 Glucose Membrane Kit
and precision may be compromised due to the software algorithm that 2776 Glucose Standard Solution (2.50 g/L)
subtracts glucose from sucrose. To avoid compromising accuracy, refer to 2777 Glucose Standard Solution (25.0 g/L)
Application Note 204LS.
2703 Sucrose Membrane Kit
2780 Sucrose Standard Solution (5.00 g/L)
2778 Sucrose Standard Solution (25.0 g/L)
2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

36 YSI Life Sciences | Application Note 216LS

 Application Note 217LS

Glucose Measurement in Corn Syrup

and Other Syrup Products
INTRODUCTION II. METHOD
Dextrose (D-glucose) concentrations in complex matrices A. Weigh 0.500 to 5.000 g of the corn syrup to be
such as corn syrup can be measured directly and quickly analyzed.
using the YSI 2900 Series Biochemistry Analyzer. YSI’s
unique enzyme technology provides for specific glucose B. Transfer the sample to a 100 mL volumetric flask, using
measurement. Measurements are virtually unaffected by buffer diluent to rinse and dilute. Fill the flask to
color, turbidity, density, pH, or the presence of reducing the mark with buffer and mix. Allow the solution to
substances. equilibrate for about twenty minutes before analysis.

When a sample is injected into the sample chamber, the C. Calibrate the 2900 Series instrument with a 2.50 g/L
glucose diffuses into the membrane containing glucose glucose standard solution.
oxidase. The glucose is immediately oxidized to hydrogen
peroxide and D-glucono-δ-lactone. The hydrogen peroxide D. Check the linearity of the membranes at least once
is detected amperometrically at the platinum electrode a day by injection of a glucose linearity check solution
surface. The current flow at the electrode is directly (9.00 g/L). Refer to the Operations Manual for
proportional to the hydrogen peroxide concentration, and specifications.
hence to dextrose concentration.
E. Assay the sample prepared in B by aspiration into the
I. MATERIALS & SETUP 2900 Series instrument. The linear range of the system is
A. YSI 2900 Series Biochemistry Analyzer - equipped 0.05 to 9.00 g/L glucose. If the value reported exceeds
with a 2365 Glucose Membrane and 2357 Buffer. this, further dilution is required.*

B. Glucose standards (2.50 g/L, 9.00 g/L). F. Calibrate frequently as described in the Operations
Manual.
C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
in reagent water). * The glucose linearity on the 2900 Series may be increased to
0.05 to 25.0 g/L. This can be done by decreasing the sample size
D. Connect the 2900 Series instrument to a suitable to 10μL and checking the linearity with a 25.0 g/L standard.
power source.
III. CALCULATIONS
E. Perform the instrument and membrane daily checks To calculate % glucose, multiply the reported value by the
described in the Operations Manual. appropriate dilution factor.

F. Volumetric glassware (Class A recommended). Example: 2.555 g of corn syrup was diluted to 100 mL in a
Class A volumetric flask. When assayed, the value reported
G. The following instrument setup is recommended: was 4.65 g/L glucose.
Sample Size 25 μL

Probe A Parameters Autocal Parameters % Glucose: = 0.1820 g glucose/g corn syrup

Chemistry Glucose Temperature 1°C 4.65 g/L x 0.100L
/2.555 g = 18.2% (w/w)
Unit g/L Time 30 Min
Calibrator 2.50 Sample 5 Sam
End Point 30 Sec Cal Shift 2%

37 YSI Life Sciences | Application Note 217LS

 Application Note 217LS

ORDERING INFORMATION
YSI Part Numbers:
2900 Biochemistry Analyzer
2365 Glucose Membrane Kit
2776 Glucose Standard Solution (2.50 g/L)
1531 Glucose Standard Solution (9.00 g/L)
2777 Glucose Standard Solution (25.0 g/L)
2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

38 YSI Life Sciences | Application Note 217LS

 Application Note 218LS

Measurement of Glucose and Sucrose in Potatoes

INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in H. The following instrument setup is recommended:
complex matrices such as potatoes can be measured directly Sample Size 25 μL
and quickly using the YSI 2900 Series Biochemistry Analyzer.
YSI’s unique enzyme technology provides for rapid dextrose Probe A Parameters Autocal Parameters
and sucrose measurement. Measurements are virtually Chemistry Glucose Temperature 1°C
unaffected by color, turbidity, density, pH, or the presence Unit g/L Time 30 Min
of reducing substances. Calibrator 2.50 Sample 5 Sam
End Point 30 Sec Cal Shift 2%
When a YSI 2900 Series Biochemistry Analyzer is equipped
with a dextrose membrane, both sucrose and dextrose II. METHOD
concentrations can be measured. This is accomplished by A. Weigh 100 to 200 grams of washed and peeled
first determining the glucose concentration. The sucrose potatoes. For information on sample selection, see J.
is then hydrolyzed to dextrose, and the total dextrose R. Sowokinos, American Potato Journal, 50, 333-334
concentration is measured. The difference in the responses (1978).
corresponds to the sucrose concentration.
B. Juicerate the potatoes in an Acme Juicerator and collect
After a sample is injected into the sample chamber, the the juice in a beaker. Wash the juicerator three times
dextrose diffuses to the glucose membrane, which contains with 100 mL portions of buffer diluent. Wait two to three
glucose oxidase, and is oxidized to hydrogen peroxide and minutes between washings.
D-glucono-δ-lactone. The hydrogen peroxide is detected
amperometrically at the platinum electrode. The current C. Quantitatively transfer the combined juice and buffer
produced is directly proportional to the hydrogen peroxide to a 500 mL volumetric flask. Rinse the beaker with
and dextrose concentrations. For more information on this several small (10 mL) aliquots of buffer and transfer to
system, refer to the Operations Manual. the flask. Dilute to the mark with buffer. Refrigerate for
one hour prior to analysis.*
I. MATERIALS & SETUP
A. YSI 2900 Series Biochemistry Analyzer - equipped D. Remove about 3 mL of the solution from C and
with a 2365 Glucose Membrane and 2357 Buffer. add ~2 mg of invertase enzyme. Stir gently until
dissolved. Cover the sample and allow incubation at
B. Glucose standards (2.50 g/L, 9.00 g/L). room temperature for 20 minutes before analysis.

C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4

E. Calibrate the 2900 series instrument witha 2.50 g/L
in reagent water).
glucose standard solution.

D. Invertase - Sigma Chemical Company I-4504

F. Check the linearity of the membrane at least once a
recommended.
day by injection of a glucose linearity check solution
(9.00 g/L). Refer to the Operations Manual for
E. Connect the 2900 Series instrument to a suitable
specifications.
power source.

G. Assay the sample prepared in C by aspiration into the

F. Perform the instrument and membrane daily checks
2900 Series. This is the free glucose concentration
described in the Operations Manual.
(Dfree).**
G. Volumetric glassware (Class A recommended).

39 YSI Life Sciences | Application Note 218LS

 Application Note 218LS

H. Assay the sample prepared in D (with invertase). The

value reported is the sum of the free glucose and that
produced from sucrose hydrolysis (Dtotal).
% Sucrose:
I. Calibrate frequently as described in the Operations (3.36 g/L - 2.13 g/L) x 0.500 L x 342.30 g/mole sucrose
Manual.
223 g 180.16 g/mole dextrose
* For potato samples with low glucose and sucrose levels, consider = 0.00524 g sucrose/g potatoes
increasing the ratio of potato sample to the volume of extracting buffer,
or consider increasing the sample size aspirated into the instrument (II.H). = 0.52% (w/w)

** The linear range of the 2900 Series instrument may be increased to 25.0
g/L. This can be done by decreasing the sample size to 10 μL and checking
the linearity with a 25.0 g/L standard.

III. CALCULATIONS
Example: A 223 g potato sample was prepared as described
in II. When the sample from II.C was assayed, the value
reported (Dfree) was 2.13 g/L glucose. The value reported for
the sample from II.D (with invertase) was 3.36 g/L dextrose
(Dtotal).

To calculate % glucose, multiply the reported value (Dfree) ORDERING INFORMATION

by the appropriate dilution factor. YSI Part Numbers:
2900 Biochemistry Analyzer
To calculate % sucrose, subtract Dfree from Dtotal and
2365 Glucose Membrane Kit
multiply by the appropriate dilution and mass ratio factors.
2776 Glucose Standard Solution (2.50 g/L)
2777 Glucose Standard Solution (25.0 g/L)
% Glucose: 1531 Glucose Standard Solution (9.00 g/L)
= 0.00478 g glucose/g potatoes
2.13 g/L x 0.500 2357 Buffer Kit
L/223 g = 0.48% (w/w) 2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

40 YSI Life Sciences | Application Note 218LS

 Application Note 219LS

Dextrose Measurement in Potatoes

INTRODUCTION II. METHOD
Dextrose (D-glucose) concentrations in complex matrices A. Weigh 100 to 200 grams of washed and peeled
such as potatoes can be measured directly and quickly potatoes. For information on sample selection, see J.
using the YSI 2900 Series Biochemistry Analyzer. YSI’s R. Sowokinos, American Potato Journal, 50, 333-334
unique enzyme technology provides for specific dextrose (1978).
measurement. Measurements are virtually unaffected by
color, turbidity, density, pH, or the presence of reducing B. Juicerate the potatoes in an Acme Juicerator and collect
substances. the juice in a beaker. Wash the juicerator three times
with 100 mL portions of buffer diluent. Wait two to three
When a sample is injected into the sample chamber, the minutes between washings.
dextrose diffuses into the membrane containing glucose
oxidase. The dextrose is immediately oxidized to hydrogen C. Quantitatively transfer the combined juice and buffer
peroxide and D-glucono-δ-lactone. The hydrogen peroxide to a 500 mL volumetric flask. Rinse the beaker with
is detected amperometrically at the platinum electrode several small (10 mL) aliquots of buffer and transfer to
surface. The current flow at the electrode is directly the flask. Dilute to the mark with buffer. Refrigerate for
proportional to the hydrogen peroxide concentration, and one hour prior to analysis.*
hence to dextrose concentration.
D. Calibrate the 2900 series instrument with a 2.50 g/L
I. MATERIALS & SETUP dextrose standard solution.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2365 Dextrose Membrane and 2357 Buffer. E. Check the linearity of the membrane at least once a
day by injection of a dextrose linearity check solution
B. Dextrose standards (2.50 g/L, 9.00 g/L). (9.00 g/L). Refer to the Operations Manual for
specifications.
C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
in reagent water). F. Assay the sample prepared in C by aspiration into the
2900 Series. The linear range of the system is 0.05 to
D. Connect the 2900 Series instrument to a suitable 9.00 g/L dextrose. If the value reported exceeds this,
power source. further dilution is required.*

E. Perform the instrument and membrane daily checks G. Calibrate frequently as described in the Operations
described in the Operations Manual. Manual.

F. Volumetric glassware (Class A recommended). * For potato samples with low dextrose content, consider increasing
the ratio of potato sample to the volume of extracting buffer. For higher
G. The following instrument setup is recommended: dextrose levels, more dilute samples are recommended.
Sample Size: 25 μL
The dextrose linearity of the 2900 series may be increased to 0.05 to 25.0
g/L. This can be done by decreasing the samples size to 10 μL and checking
Probe A Parameters Autocal Parameters the linearity with a 25.0 g/L standard.
Chemistry Glucose Temperature 1°C
Unit g/L Time 30 Min
Calibrator 2.50 Sample 5 Sam
End Point 30 Sec Cal Shift 2%

41 YSI Life Sciences | Application Note 219LS

 Application Note 219LS

III. CALCULATIONS ORDERING INFORMATION

To calculate % dextrose, multiply the reported value by the YSI Part Numbers:
appropriate dilution factor. 2900 Biochemistry Analyzer
2365 Glucose Membrane Kit
Example: A 200 g potato sample was prepared as de- 2776 Glucose Standard Solution (2.50 g/L)
scribed in III.B and C. When assayed, the value reported 1531 Glucose Standard Solution (9.00 g/L)
was 2.15 g/L dextrose. 2777 Glucose Standard Solution (25.0 g/L)
2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
% Dextrose: = 0.00538 g dextrose/g 2392 NaCl Solution (for membrane installation) n
2.15 g/L x 0.500 potatoes
L/200 g = 0.54% (w/w)

42 YSI Life Sciences | Application Note 219LS

 Application Note 220LS

Simultaneous Measurement of Dextrose

and Sucrose in Molasses
INTRODUCTION
Dextrose (D-glucose) and sucrose concentrations in C. Buffer Diluent (40 g/L NaH 2PO 4, 10g/L Na 2HPO 4
complex matrices such as molasses can be measured in reagent water).
directly and quickly using the YSI 2900 Series Biochemistry
Analyzer. YSI’s unique enzyme technology provides for rapid D. Connect the 2900 Series instrument to a suitable power
dextrose and sucrose measurements. Measurements are source.
virtually unaffected by color, turbidity, density, pH, or the
presence of reducing substances. E. Perform the instrument and membrane daily checks
described in the Operations Manual.
When a 2900 Series Biochemistry Analyzer is equipped
with a dextrose and a sucrose membrane, simultaneous F. Volumetric glassware (Class A recommended).
measurement of both analytes is possible. Because dextrose
interferes with sucrose analysis, it is necessary to follow G. The injection volume should be set to 25µL.
this protocol when analyzing for sucrose in the presence
of dextrose. H. The following instrument setup is recommended.

When a sample is injected into the sample chamber, the Probe A Parameters Probe B Parameters
sucrose diffuses to the sucrose membrane, which contains Chemistry Glucose Chemistry Sucrose
invertase, mutarotase, and glucose oxidase. The sucrose is Unit g/L Unit g/L
hydrolyzed to α-D-glucose and fructose. The mutarotase Calibrator 2.50 Calibrator 5.00 g/L
allows for the quick equilibrium of glucose between End Point 30 Sec End Point 30 Sec
the α and β forms. In the presence of glucose oxidase,
the β-D-glucose is oxidized to hydrogen peroxide and Autocal Parameters
D-glucono-δ-lactone. The hydrogen peroxide is detected Temperature 1°C
amperometrically at the platinum electrode surface. The Time 30 Min
dextrose in the sample diffuses to both the dextrose and Sample 5 Sam
sucrose membranes, which contain glucose oxidase, and is Cal Shift 2%
oxidized as well. Subtracting the response of the dextrose
membrane from the response of the sucrose membrane
yields the response due to sucrose alone. The dextrose II. METHOD
response is taken directly from the dextrose membrane. A. Weigh up to 5.000 g of molasses to be analyzed.
The algorithm in the instrument software calculates the net
concentrations. For more information on this system, refer B. Transfer the sample to a 100 mL volumetric flask using
to the Operations Manual. buffer diluent to rinse and dilute. Fill the flask to the
mark with buffer and mix. Allow the solution to
I. MATERIALS & SETUP equilibrate for about twenty minutes before analysis.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2703 Sucrose Membrane, a 2365 Dextrose C. Calibrate the 2900 Series instrument with 2.50 g/L
Membrane and 2357 Buffer. dextrose and 5.00 g/L sucrose standard solutions.

B. Dextrose (2.50 g/L, 9.00 g/L) and Sucrose (5.00 g/L, D. Check the linearity of the membrane at least once
25.0 g/L) standard solutions. a day by injection of dextrose (9.00 g/L) and sucrose

43 YSI Life Sciences | Application Note 220LS

 Application Note 220LS

linearity check solutions (25.0 g/L). Refer to the

Operations Manual for specifications. % Dextrose: = 0.1477 g dextrose/g molasses
6.75 g/L x 0.100L
/4.569 g = 14.8% (w/w)
E. Assay the sample prepared in B by aspiration into the
2900 Series instrument.*

F. Calibrate frequently as described in the Operations

% Sucrose: = 0.2832 g sucrose/g molasses
Manual.
12.94 g/L x 0.100L
/4.569 g = 28.3% (w/w)
* The combined concentration of dextrose + sucrose cannot exceed 25
g/L. If the sum of the values reported exceeds this, further dilution of the
sample is required.
ORDERING INFORMATION
If the dextrose concentration exceeds the sucrose concentration, accuracy
and precision may be compromised due to the software algorithm that
YSI Part Numbers:
subtracts dextrose from sucrose. To avoid compromising accuracy, refer 2900 Biochemistry Analyzer
to Application Note 204LS. 2365 Dextrose Membrane Kit
2776 Dextrose Standard Solution (2.50 g/L)
III. CALCULATIONS 1531 Dextrose Standard Solution (9.00 g/L)
To calculate % dextrose and sucrose, multiply the values 2703 Sucrose Membrane Kit
reported by the appropriate dilution factor. 2780 Sucrose Standard Solution (5.00 g/L)
2778 Sucrose Standard Solution (25.0 g/L)
Example: 4.569 g of molasses was diluted to 100 mL in 2357 Buffer Kit
a Class A volumetric flask. When assayed, the values 2363 Potassium Ferrocyanide Test Solution
reported were 6.75 g/L dextrose and 12.94 g/L sucrose. 2392 NaCl Solution (for membrane installation) n

44 YSI Life Sciences | Application Note 220LS

 Application Note 221LS

Sucrose Measurement in Molasses

INTRODUCTION
Sucrose concentrations in complex matrices such as F. The following instrument setup is recommended.
molasses can be measured directly and quickly using the Sample size 25 μL
YSI 2900 Series Biochemistry Analyzer. YSI’s unique enzyme
technology provides for rapid sucrose measurement. Probe A Parameters Autocal Parameters
Measurements are virtually unaffected by color, turbidity, Chemistry Sucrose Temperature 1°C
density, pH, or the presence of reducing substances. Unit g/L Time 30 Min
Calibrator 5.00 Sample 5 Sam
When a sample is injected into the sample chamber, the End Point 30 Sec Cal Shift 2%
sucrose diffuses into the membrane containing invertase,
mutarotase, and glucose oxidase. The sucrose is hydrolyzed
to α-D-glucose and fructose. The mutarotase allows for the II. METHOD
quick equilibrium of glucose between its α and β forms. In A. Weigh up to 5.000 g of molasses to be analyzed.
the presence of glucose oxidase, the β-D-glucose (dextrose)
is immediately oxidized to hydrogen peroxide and B. Transfer the sample to a 100 mL volumetric flask
D-glucono-δ-lactone. The hydrogen peroxide is detected using buffer diluent to rinse and dilute. Fill the flask to
amperometrically at the platinum electrode surface. The the mark with buffer and mix.
current flow at the electrode is directly proportional to the
hydrogen peroxide concentration, and through the series C. Calibrate the 2900 Series instrument with a 5.00 g/L
of reactions described above, the hydrogen peroxide sucrose standard solution.
concentration is also directly proportional to the sucrose
concentration. D. Check the linearity of the membrane at least once
a day by injection of sucrose linearity check solutions
Because the membrane contains glucose oxidase, any (25.00 g/L). Refer to the Operations Manual for
dextrose in the sample will also be oxidized and produce specifications.
a signal. For this reason, the sample must be dextrose-free.
If dextrose is present in the sample, refer to Application E. Assay the sample prepared in B by aspiration into the
Note 220LS for the Simultaneous Measurement of Dextrose 2900 Series. The linear range of the system is 0.1 to
and Sucrose in Molasses. For more information, refer to the 25.00 g/L sucrose. If the value reported exceeds this,
Operations Manual. further dilution is required.

I. MATERIALS & SETUP F. Calibrate frequently as described in the Operations

A. YSI 2900 Series Biochemistry Analyzer - equipped Manual.
with a 2703 Sucrose Membrane and 2357 Buffer.
III. CALCULATIONS
B. Sucrose standards (5.00 g/L25.0 g/L). To calculate % sucrose, multiply the reported value by the
appropriate dilution factor.
C. Connect the 2900 Series instrument to a suitable power
source. Example: 2.001 g of molasses was diluted to 100 mL in a
Class A volumetric flask. When assayed, the value reported
D. Perform the instrument and membrane daily checks was 5.89 g/L sucrose.
described in the Operations Manual.

E. Volumetric glassware (Class A recommended).

45 YSI Life Sciences | Application Note 221LS

 Application Note 221LS

ORDERING INFORMATION
% Sucrose: = 0.294 g sucrose/g molasses YSI Part Numbers:
5.89 g/L x 0.100L 2900 Biochemistry Analyzer
/2.001 g = 29.4% (w/w)
2703 Sucrose Membrane Kit
2780 Sucrose Standard Solution (5.00 g/L)
2778 Sucrose Standard Solution (25.0 g/L)
2357 Buffer Kit
2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

46 YSI Life Sciences | Application Note 221LS

 Application Note 222LS

Determination of % Cook in Extruded Cereal Products

INTRODUCTION
The degree of cook of extruded cereal products can I. Perform the instrument and membrane daily checks
be determined using the YSI 2900 Series Biochemistry described in the Operations Manual.
Analyzer. YSI’s unique enzyme technology provides for
specific glucose measurement. Measurements are virtually J. The following instrument setup is recommended:
unaffected by color, turbidity, density, pH or the presence Sample Size 25 μL
of reducing substances.
Probe A Parameters Autocal Parameters
A portion of a sample is solubilized in cold water and a Chemistry Glucose Temperature 1°C
portion is autoclaved. The samples containing starch are Unit g/L Time 30 Min
treated identically with glucoamylase. Glucose produced Calibrator 2.50 Sample 5 Sam
from this reaction is measured with the YSI 2900 Series End Point 30 Sec Cal Shift 2%
instrument. The ratio of glucose in the cold water sample to
glucose in the autoclaved sample yields % cook. II. METHOD
A. Cold water sample:
When a sample is injected into the sample chamber, the 1. Into a 125 mL Erlenmeyer flask disperse ~ 0.50 g
glucose diffuses into the membrane containing glucose sample into ~ 43 mL reagent water. Record the exact
oxidase. The glucose is immediately oxidized to hydrogen weight of the sample.
peroxide and D-glucono-δ-lactone. The hydrogen peroxide
is detected amperometrically at the platinum electrode 2. Add 5 mL of 1N Acetate buffer, pH 4.2.
surface. The current flow at the electrode is directly
proportional to the hydrogen peroxide concentration, and 3. Add 2.5 mL of 30% glucoamylase solution
hence to the glucose concentration.
4. Cover with aluminum foil and incubate, in water
I. MATERIALS & SETUP bath, for one hour at 40°C.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2365 Glucose Membrane and 2357 Buffer. 5. Add ~ 3.2 mL of 25% Trichloracetic Acid immediately
after the incubation and swirl the contents.
B. Glucose standards (2.50 g/L, 9.00 g/L).
6. Allow the solution to cool to room temperature.
C. 1N Acetate buffer. Transfer the solution to a 100 mL volumetric flask
and dilute with phosphate diluent buffer, pH 5.9.
D. Glucoamylase solution. Shake vigorously.

E. 25% Trichloracetic Acid.

B. Autoclaved sample:
1. Dilute the sample as in A1. Cover with aluminum
F. Phosphate diluent buffer (40 g/L NaH2PO4, 10 g/L
foil and autoclave at 15-20 psi, ~124°C ± 3°C for
Na2HPO4 in reagent water, pH 5.9).
one hour. Cool to 40°C.
G. Volumetric glassware (Class A recommended).
2. Repeat steps A2 - A6 above.
H. Connect the 2900 Series instrument to a suitable
power source.

47 YSI Life Sciences | Application Note 222LS

 Application Note 222LS

C. Blank sample:
[Cooked Starch]
Since glucoamylase may contain free glucose, perform % Cook = x 100%
steps A1-A6 without using the sample containing starch. [Total Starch]
Both the cold water sample and the autoclaved sample or
should be corrected using this value. [(Step G - Step F) x 0.9]
% Cook = x 100%
D. Calibrate the 2900 Series instrument with a 2.50 [(Step H - Step F) x 0.9]
g/L glucose standard solution.

E. Check the linearity of the membrane at least once a Example: 0.52 g of pet food was diluted to 100 mL in a
day by injection of a glucose linearity check solution Class A volumetric flask. The sample was prepared using
(9.00 g/L). Refer to the Operators Manual for the cold water procedure. When assayed, the value report-
specifications. ed was 1.45 g/L glucose.

F. Determination of Blank: Assay the blank prepared in C 0.52 g of pet food was diluted to 100 mL in a Class A vol-
by aspiration into the 2900 Series instrument.* umetric flask. The sample was prepared using the auto-
claved procedure. When assayed, the value reported was
G. Determination of Cooked Starch: Assay the sample 1.82 g/L glucose.
prepared in A by aspiration into the 2900 Series
Instrument.* The blank contained 0.01 g/L of glucose.

H. Determination of Total Starch: Assay the sample Cold water starch: = 0.249 g starch/g food
prepared in B by aspiration into the 2900 Series 1.45-0.01 g/L x 0.9 x
0.100L/0.52 g = 24.9% (w/w)
Instrument.*

I. Calibrate frequently as described in the Operations

Manual. Total starch: = 0.313 g starch/g food
1.82-0.01 g/L x 0.9 x
* The linear range of the system is 0 to 9.00 g/L glucose. If the value 0.100L/0.52 g = 31.3% (w/w)
reported exceeds this, further dilution is required.

Note: If the sample contains free glucose, both the cold water and
the autoclaved sample will have to be corrected with this value.
% Cook:
Weigh 0.5 grams of sample into 100 mL volumetric flask and dilute = 79.6%
to the mark with phosphate diluent buffer. Mix the sample until 24.9% / 31.3% x 100%
dissolved and analyze.

III. CALCULATIONS ORDERING INFORMATION

To calculate % cook, multiply the reported value by the YSI Part Numbers:
appropriate dilution factor. The value of the blank (measured 2900 Biochemistry Analyzer
step F) should be subtracted from the cooked starch 2365 Glucose Membrane Kit
(measured in step G) and the total starch (measured in 2776 Glucose Standard Solution (2.50 g/L)
step H). 1531 Glucose Standard Solution (9.00 g/L)
2357 Buffer Kit
Since 1.1 g of glucose is produced when 1.0 g of starch is 2363 Potassium Ferrocyanide Test Solution
hydrolyzed, the glucose concentration of the sample should 2392 NaCl Solution (for membrane installation) n
be multiplied by 0.9.

48 YSI Life Sciences | Application Note 222LS

 Application Note 223LS

Determination of % Cook in Extruded Cereal

Products Using Chemical Solubilization
INTRODUCTION
The degree of cook of extruded cereal products can I. Phosphate buffer (40 g/L NaH2PO4, 10 g/L Na2HPO4
be determined using the YSI 2900 Series Biochemistry in reagent water).
Analyzer. YSI’s unique enzyme technology provides for
specific glucose measurement. Measurements are virtually J. Connect the 2900 Series instrument to a suitable power
unaffected by color, turbidity, density, pH, or the presence source.
of reducing substances.
K. Perform the instrument and membrane daily checks
A portion of a sample is solubilized in cold water and a described in the Operations Manual.
portion is autoclaved or chemically solubilized. The samples
containing starch are treated identically with glucoamylase. L. Volumetric glassware (Class A recommended).
The glucose produced from this reaction is measured with
the YSI 2900 Series. In this procedure chemical solubilization M. The following instrument setup is recommended:
is described. See Application Note 222LS for the autoclaved Sample Size 25 μL
method. The ratio of glucose in the cold water sample to
glucose in the chemically solubilized sample yields % cook. Probe A Parameters Autocal Parameters
Chemistry Glucose Temperature 1°C
When a sample is injected into the sample chamber, the Unit g/L Time 30 Min
glucose diffuses into the membrane containing glucose Calibrator 2.50 Sample 5 Sam
oxidase. The glucose is immediately oxidized to hydrogen End Point 30 Sec Cal Shift 2%
peroxide and D-glucono-δ-lactone. The hydrogen peroxide
is detected amperometrically at the platinum electrode II. REAGENT PREPARATION
surface. The current flow at the electrode is directly A. Acetate Buffer (pH 4.2) - Weigh 9.1 grams of sodium
proportional to the hydrogen peroxide concentration, and acetate into 500 mL volumetric flask. Add about 300
hence to the glucose concentration. mL of distilled water and mix until the entire solid is
dissolved. Add 22.3 mL (23.4 grams) of glacial acetic
I. MATERIALS & SETUP acid. Dilute to volume with distilled water and mix.
A. YSI 2900 Series Biochemistry Analyzer - equipped
with a 2365 Glucose Membrane and 2357 Buffer. B. Glucoamylase Enzyme Solution - Pipette 30 mL of
glucoamylase into a 100 mL volumetric flask. Add 0.1
B. Glucose standards (2.5 g/L, 9.00 g/L). gram of EDTA (Ethylenediaminetetraacetic acid) and
dilute to volume with distilled water. Mix thoroughly to
C. 1N Acetate buffer. dissolve the EDTA and the glucoamylase.

D. Glucoamylase solution. C. Hydrochloric Acid Solution (2N) - For example: Measure

82.4 mL of 36.5-38% hydrochloric and transfer to a
E. 25% Trichloracetic Acid. 500 mL volumetric flask. Let cool, dilute to volume with
distilled water and mix.
F. 2N Sodium Hydroxide.
D. Sodium Hydroxide Solution (2N) - For example: Weigh
G. 2N Hydrochloric Acid.
40 grams of sodium hydroxide pellets into a 500 mL
volumetric flask. Add 300 mL of distilled water and mix.
H. A heating unit such as a hot plate power source.
Let cool, dilute to volume and mix.

49 YSI Life Sciences | Application Note 223LS

 Application Note 223LS

E. Trichloroacetic Acid Solution (25%) - Dissolve 50.0 Blank sample

grams of TCA crystals into 200 mL of distilled water.
J. Since glucoamylase may contain free glucose, perform
III. METHOD steps F-I without using the sample containing starch.
A. Grind sample to a fine powder. Both the cold water sample and the autoclaved sample
should be corrected using this value.
B. Weigh out 0.50 grams of sample twice and transfer
each to a 100 mL volumetric flask. Record exact weights. K. Calibrate the 2900 Series instrument with a 2.50 g/L
glucose standard solution.
C. Add 25 mL of distilled water to each flask. Label one
flask #1 and the second flask #2. To the flask labeled #1 L. Check the linearity of the membrane at least once a day
proceed with the chemical solubilization. Set flask #2 by injection of a glucose linearity check solution (9.00
aside until the enzymatic digestion in steps F-I. g/L). Refer to the Operators Manual for specifications.

Chemical solubilization to determine total starch M. Determination of Glucose: Assay the blank prepared
in J by aspiration into the 2900 Series instrument.*
D. Add 10 mL 2N sodium hydroxide to the solution in flask
#1. Place on a heating unit and simmer for 20 minutes. N. Determination of Cooked Starch: Assay the sample
Stir gently and periodically. prepared in flask #2 by aspiration into the 2900 Series
instrument.*
E. Add 10 mL 2N hydrochloric acid following the 20
minutes and swirl the flask. Allow the flask to cool to O. Determination of Total Starch: Assay the sample
below 50°C. prepared in flask #1 by aspiration into the 2900 Series
instrument.*
Enzymatic digestion to determine cooked starch
P. Calibrate frequently as described in the Operations
F. To both flasks (#1 and #2) add 10 mL of 1N acetate Manual.
buffer.
* The linear range of the system is 0.05–9.00 g/L glucose. If the value
reported exceeds this, further dilution is required.
G. Add 5 mL of 30% glucoamylase solution to each flask.
Mix well and place the flask in a 40°C water bath for If the sample contains free glucose, both the cold water and the
70 minutes. autoclaved sample will have to be corrected with this value. Weigh
0.5 grams of sample into a 100 mL volumetric flask and dilute to
H. After exactly 70 minutes incubation, remove the flasks the mark with phosphate buffer. Mix the sample for 20 minutes
and analyze.
from the water bath. Immediately add 5 mL of 25% TCA
to each flask to stop hydrolysis.
VI. CALCULATIONS
I. Cool to room temperature and fill to volume with To calculate % cook, multiply the reported value by the
phosphate diluent buffer and mix well. appropriate dilution factor. The value of the blank (measured
in step J) should be subtracted from the cooked starch
(measured in step N) and the total starch (measured in
step O).

50 YSI Life Sciences | Application Note 223LS

 Application Note 223LS

Since 1.1 g of glucose is produced when 1.0 g of starch is

hydrolyzed, the glucose concentration of the sample should
be multiplied by 0.9.

[Cooked Starch]
% Cook = x 100%
[Total Starch]
or
[(Step N - Step J) x 0.9]
% Cook = x 100%
[(Step O - Step J) x 0.9]

Example: 0.52 g of pet food was diluted to 100 mL in a

Class A volumetric flask. The sample was prepared using
the enzymatic digestion procedure. When assayed, the
value reported was 1.45 g/L glucose.

A 0.52 g of pet food was diluted to 100 mL in a Class

A volumetric flask. The sample was prepared using the
chemical solubilization procedure. When assayed, the
value reported was 1.82 g/L glucose.

The blank contained 0.01 g/L of glucose.

Cooked starch: = 0.249

1.45 - 0.01 g/L x 0.9 x 0.100L/0.52 g = 24.9 %

= 0.313 ORDERING INFORMATION

Total starch:
YSI Part Numbers:
1.82 - 0.01 g/L x 0.9 x 0.100L/0.52 g = 31.3% 2900 Biochemistry Analyzer
2365 Glucose Membrane Kit
2776 Glucose Standard Solution (2.50 g/L)
1531 Glucose Standard Solution (9.00 g/L)
% Cook = 2357 Buffer Kit
= 79.6%
0.249 / 0.313 x 100% 2363 Potassium Ferrocyanide Test Solution
2392 NaCl Solution (for membrane installation) n

51 YSI Life Sciences | Application Note 223LS

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