YSI 2700 SELECT Biochemistry Analyzer

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Garry Pitts
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1 YSI 2700 SELECT Biochemistry Analyzer

2 Immobilized Enzyme Biosensor Key to accurate measurement YSI biosensor technology is the key to fast, accurate measurement. What does this mean for you? Results that are specific for the analyte of interest Fast results in 60 seconds results you can use to adjust your process Low operating costs enzymes are used over and over An enzyme specific for the substrate of interest is immobilized between two membrane layers: polycarbonate and cellulose acetate. The substrate is oxidized as it enters the enzyme layer, producing hydrogen peroxide, which passes through cellulose acetate to a platinum electrode, where the hydrogen peroxide is oxidized. The resulting current is proportional to the concentration of the substrate. YSI membranes contain three layers. The first layer, porous polycarbonate, limits the diffusion of the substrate into the second enzyme layer, preventing the reaction from becoming enzymelimited. The third layer, cellulose acetate, permits only small molecules, such as hydrogen peroxide, to reach the electrode, eliminating many electrochemically-active compounds that could interfere with the measurement. 2 Analyte Oxygen Hydrogen peroxide GLUCOSE MEASUREMENT D-glucose is oxidized, producing hydrogen peroxide and glucono-lactone. Polycarbonate layer An example of the reaction that takes place in a YSI immobilized-enzyme biosensor. D-glucose + O2 SUCROSE MEASUREMENT requires three immobilized enzymes: invertase, mutarotase and glucose oxidase. The invertase converts sucrose to glucose and fructose. Mutarotase allows the rapid conversion of a-d-glucose to b-d-glucose until an equilibrium is established. Then b-d-glucose is oxidized in the presence of glucose oxidase, producing hydrogen peroxide. We make immobilized-enzyme biosensors to measure these substrates: Glucose Sucrose Lactose Lactate Galactose Glutamate Choline Glutamine Ethanol Sucrose + H2O α-d-glucose β-d-glucose + O2 Immobilized enzyme layer glucose oxidase glucose oxidase Current (nanoamperes) invertase mutarotase Cellulose acetate layer D-glucono-δ-lactone + H202 α-d-glucose +(fructose) β-d-glucose Platinum electrode D-glucono-δ-lactone + H2O Time (seconds) TYPICAL BIOSENSOR RESPONSE CURVE Once the substrate reaches the sensor, current develops rapidly, reaching a steady-state in 15 to 30 seconds. The current is directly proportional to the substrate concentration in the sample.

Food Analysis IN many instances, measuring key components during the manufacture of food products can prevent the loss of batches due to misformulation.

The YSI SELECT can meet your needs for real-time, accurate analysis of key food components during manufacturing.

3 Food Analysis IN many instances, measuring key components during the manufacture of food products can prevent the loss of batches due to misformulation. Traditional methods, such as HPLC and GC, require high maintenance, expert operators, and long analysis times, making them impractical for process monitoring. The YSI SELECT can meet your needs for real-time, accurate analysis of key food components during manufacturing. You can use the analyzer to measure common food ingredients, such as: Glucose (Dextrose) Sucrose Lactose Lactate Galactose Glutamate Choline Glutamine Ethanol Hydrogen Peroxide When placed in a manufacturing facility, the analyzer, with realtime results, allows immediate verification of formulation or immediate intervention and reformulation, if necessary. Because the instrument is simple to use, extensive operator training is not required, allowing a number of people to operate it with confidence. Quality checks throughout its operation guarantee reproducible, accurate results. Batch Analysis The optional YSI 2710 Turntable facilitates the analysis of batches of up to 24 samples. Load the turntable, and the YSI SELECT will automatically analyze all the samples, print the results, and send them to a PC via RS232 connection, if desired. Other Features Results are specific for your analytes, even in complex matrices Select measurement units: g/l, mg/l, mmol/l or % (w/v) Select calibration values and frequency for your application Two measurement chemistries can be run simultaneously Easy-to-follow menus prompt you when operating the unit 3

More recently, researchers have recognized the need to regulate nutrients and byproducts as well, and have determined that controlling them plays a vital role in the health and productivity of their

4 Bioprocess Monitoring FERMENTATIONS and cell cultures require tight control of system variables to achieve consistent, desirable results. Regulating variables, such as oxygen, ph and temperature, has long been accepted as important for assuring viable processes. More recently, researchers have recognized the need to regulate nutrients and byproducts as well, and have determined that controlling them plays a vital role in the health and productivity of their processes. The YSI SELECT Biochemistry Analyzer provides vital information about your processes. It provides rapid, accurate analysis of key nutrients and byproducts, including: Glucose Lactate Glutamine Glutamate Ethanol Lactose Sucrose Galactose Hydrogen peroxide Sample analysis can be done two ways, off-line or on-line. 4 Off-Line Analysis For many processes, off-line analysis is a good method for monitoring nutrients or byproducts, particularly if consumption or production rates are slow. As a benchtop analyzer, the YSI SELECT is easy to use and provides results quickly in 60 seconds. Sample volume requirements are low about 25 µl which allows you to run as many samples as you want. And the best feature of all is that no sample preparation is required no filtration and no dilution in most cases. Couple the YSI SELECT with the YSI 2710 Turntable for batch analysis. The 24-position turntable lets you load samples and walk away. The instrument will automatically analyze them, print results and send them to a PC. On-Line Analysis For processes that require frequent monitoring and regulation, on-line analysis allows you the freedom to sample automatically and adjust nutrient and byproduct concentration. The YSI 2730 Aseptic Monitoring & Control system pulls samples directly from a bioreactor, measures important nutrients and byproducts, and immediately controls the replenishment of nutrients, if needed, while maintaining the sterility of the process. All system components are sterilizable.

5 How On-Line Monitoring Works The YSI 2730 Monitoring and Control system pulls sample directly from a bioreactor and pumps it to the YSI SELECT at programmed intervals. After each analysis, an antiseptic reagent, typically a dilute hypochlorite or hydroxide solution, is flushed through the sample lines and remains there until the next cycle to assure sterility. Isolation Filter Filtrate Pump Glucose concentration g/l Glucose Regulation in Fermentation Using YSI Analog Control Elapsed Hours Concentration Consumption A yeast fermentation was run and its glucose concentration monitored frequently. The YSI Monitoring & Control system was instructed to trigger replenishment of the nutrient solution to achieve a constant glucose concentration of 2.0 g/l. The graph indicates successful regulation of the fermentation for an extended period. Over varying glucose consumption rates (right axis), the system was able to maintain a glucose concentration of 2.0 g/l (left axis) Glucose consumption in mg/min/l Bioreactor Vessel Feedstock Pump Feedstock For those bioprocesses in which loss of cells is a concern, you can connect the YSI system to an on-line filtration device. You can adjust system parameters, such as flow rate, to accommodate the needs of the filtration device. For regulation, connect the YSI SELECT to a feed pump. When nutrient concentration falls below a programmed setpoint, the YSI SELECT will signal the feed pump to add nutrient to the bioreactor. Other system variables, such as sampling interval and concentration setpoint, allow you to customize the instrument for your process needs. For processes that consume glucose quickly, for example, you can configure the system to monitor glucose every few minutes. For other processes that consume nutrients slowly, monitoring every 30 or 60 minutes may be more appropriate. Other Features RS232 and analog signal (with YSI 2730) allows data logging with a PC Closed-loop monitoring and control capability Discrete samples can be run when analyzer is acting as on-line monitor Autoclavable on-line monitor components Two measurement chemistries can be run simultaneously Selectable measurement units: g/l, mg/l, mmol/l, % (w/v) 5

6 Chemistries UNLESS otherwise stated, specifications reflect use with YSI-supplied calibration solutions and a 25 µl sample. Sample size, calibration concentration and endpoints can be changed to accommodate specific applications. Detection Calibration Linearity Precision Linearity Linearity Typical Range Point Check CV 0 to Cal to Membrane Point n=10 cal range Working point max Life Choline mg/l 175 mg/l 450 mg/l 2% ±2% ±5% 7 days D-Glucose 0-9 g/l at 25 µl 2.50 g/l 9.0 g/l at 25 µl 2% ±2% ±5% 21 days Dextrose sample size sample size 0-25 g/l at 10 µl 25 g/l at 10 µl sample size sample size Ethanol g/l 2.00 g/l 3.20 g/l 2% ±2% ±5% 5 days Galactose 2, g/l 10 days Hydrogen mg/l 21 days Peroxide 2 Lactose 0-25 g/l 5.00 g/l 25.0 g/l 2% ±2% ±5% 10 days L-Glutamate 0-10 mmol/l 5.00 mmol/l 10.0 mmol/l 2% ±2% ±5% 7 days L-Glutamine mmol/l 5.00 mmol/l 8.00 mmol/l 4% ±4% ±5% 5 days L-Lactate g/l 0.50 g/l 2.67 g/l 2% ±2% ±5% 14 days L-Lactic Acid Sucrose 0-25 g/l 5.00 g/l 25.0 g/l 2% ±2% ±5% 10 days 1. Methanol is a potential substrate interference. Specifications are for 10 µl sample. 2. The user must provide calibrator solution. 3. Lactose is a potential substrate interference. 4. Specifications are for the simultaneous measurement of glutamine and glutamate. 6

Specifications Analyzer Aspirated sample volume: User selects 5 to 65 microliters Analysis time: 60 seconds Precision: <2% cv, n=10 Linearity: ±5%, calibration value to range maximum Calibration:

7 Specifications Analyzer Aspirated sample volume: User selects 5 to 65 microliters Analysis time: 60 seconds Precision: <2% cv, n=10 Linearity: ±5%, calibration value to range maximum Calibration: User selects frequency Size: 35.6 x 25.4 x 35.6 cm, 11.4 kg (14 x 10 x 14 inches, 25 lbs) Working environment: 15 to 35 C ambient temperature, 10 to 90% relative humidity (noncondensing) Power: 110 to 120 volts, 50/60 Hz, 1.0 amp, 3-wire grounded. 220 to 240 volts, 50/60 Hz, 3-wire grounded Battery backup: Rechargeable Ni-Cad batteries to back up RAM. Monitor Sample inlet tubing: Silicone, 0.08" OD x 0.02" ID; Volume: 5.1 microliters per inch Inlet channel pump tubing: PharMed 0.13" OD x 0.035" ID Valve tubing: 0.03" ID Wasteline tubing: Silicone, 0.16" OD x 0.10" ID Nominal flow rate (inlet line): 100 to 2500 microliters per minute ±6 PSI) Sample interval: 2 minutes to 1,000 hours Turntable Sample containers: 10 to 16 mm dia, 20 to 100 mm height Sample wheel: 19.1 dia x 1.9 cm (7.5 dia x 0.75 in) Size: 23.5 x 20.3 x 12.7 cm, 2.5 kg (9.25 x 8 x 5 in, 5.5 lbs) 7

Application Notes APPLICATION notes are available for a variety of products that our chemists have tested. Our applications support team is ready to assist you with your sample applications.

8 Application Notes APPLICATION notes are available for a variety of products that our chemists have tested. Our applications support team is ready to assist you with your sample applications. Call us at Fax No. 301 Dextrose in Molasses No. 302 Sucrose in Molasses No. 303 Simultaneous Dextrose & Sucrose in Molasses No. 304 Dextrose in Potatoes No. 305 Simultaneous Dextrose & Sucrose in Potatoes No. 306 Dextrose in Corn Syrup & Other Syrup Products No. 307 Simultaneous Dextrose & Sucrose in Sweetened Condensed Milk No. 308 Simultaneous Dextrose & Sucrose in Baked Goods No. 309 Simultaneous Dextrose & Sucrose in Cereal Products No. 310 Simultaneous Dextrose & Sucrose in Corn & Peas No. 311 L-Lactate in Lunch Meats No. 312 Dextrose in Frozen Green Beans No. 313 Ethanol in Beer No. 314 Dextrose in Canned Green Beans No. 315 Simultaneous Dextrose & Sucrose Utilizing External Hydrolysis No. 316 Simultaneous Dextrose & Sucrose in Ice Cream Bars No. 317 Determination of Hydrogen Peroxide No. 318 L-Glutamate Determination No. 319 Determination of % Cook in Extruded Cereal Products Using Physical Solubilization No. 320 Lactose Measurement in Cheese No. 322 % Cook in Extruded Cereal Products Using Chemical Solubilization No. 323 Simultaneous Dextrose & Sucrose in Peanut Butter No. 324 Choline Determination No. 325 Simultaneous Measurement of Glutamine & Glutamate Systems Application Notes No. 201 Data Logging No. 202 Effluent Monitoring No. 203 Aseptic Monitoring & Control 8

9 Typical Results YSI immobilized-enzyme biosensors provide accurate results, which the representative data on this page demonstrate. Contact us if you need to know how the YSI analyzer works with your sample. GLUTAMATE: spike & recovery Product Recovery Samples Chicken broth 100.1% 6 Salad dressing 100.0% 3 Chow mein 99.7% 3 Spice mix 99.0% 3 Tomatoes 99.7% 3 To evaluate the accuracy of our glutamate measurement, each sample was spiked with a known concentration of glutamate. In each case, nearly 100% of the spike was recovered, indicating no significant matrix interferences. CHOLINE: spike & recovery Product Spike Recovery Infant 12% 105% formula 35% 98.9% 60% 98.6% Infant 50% 99.1% formula 65% 96.1% with protein 75% 96.4% hydrolysate 100% 98.8% To evaluate the accuracy of our choline measurement, each sample was spiked with a known concentration of choline. In each case, nearly 100% of the spike was recovered, indicating no significant matrix interferences. BREAKFAST CEREALS: YSI analyzer versus HPLC YSI Analyzer % w/w HPLC % w/w Sample Sucrose Dextrose Sucrose Dextrose Label* A B C D E F G H I *Label information reported as sucrose and other sugars. The sucrose and dextrose concentrations of 9 breakfast cereals correlated well when measured with a YSI analyzer and HPLC. GLUTAMINE IN CELL CULTURE: YSI analyzer versus HPLC YSI (mmol/l) HPLC (mmol/l) The glutamine concentration of a cell culture was measured with a YSI analyzer and with HPLC. Although the results correlated well, the YSI results were available right away for process adjustments. 9

10 How to Order WE sell the YSI 2700 SELECT worldwide. For more information, please contact us. YSI United States Fax YSI Limited United Kingdom 44 (0) Fax 44 (0) YSI Hong Kong Fax YSI Japan Fax YSI SELECT Biochemistry Analyzer YSI 2700D Dual-Channel Biochemistry Analyzer YSI 2700S Single-Channel Biochemistry Analyzer YSI 2700M Dual-Channel Biochemistry Analyzer with YSI 2730 Aseptic Monitoring & Control Accessory YSI 2710 Turntable YSI 2730 Aseptic Monitoring & Control Accessory Other Supplies YSI 2788 Preventive Maintenance Kit YSI 2751 Printer Paper YSI 2392 Sodium Chloride Solution YSI 2363 Potassium Ferrocyanide Solution YSI Reagents Chemistry Membrane Buffer Calibration Linearity Check Kit Solution Solution Choline mg/l mg/l Glucose g/l g/l (25 µl sample) g/l (10 µl sample) Ethanol g/l g/l Glutamate mmol/l mmol/l Glutamine mmol/l mmol/l L-Lactate g/l g/l, g/l Lactose g/l g/l Sucrose g/l g/l In addition to the reagents listed above, you ll need YSI 2392 Sodium Chloride Solution and YSI 2363 Potassium Ferrocyanide Solution (not needed for lactose). 1. Ethanol standards supplied as a kit of 3 each 2.00 g/l and 2 each 3.20 g/l. 10

References S.K. Bhattacharya, A.K. Dubey, High-Level Expression of a Heterologous Gene in Escherichia coli in Response to Carbon-Nitrogen Source and C/N Ratio in a Batch Bioreactor, Biotechnol. Prog.

11 References S.K. Bhattacharya, A.K. Dubey, High-Level Expression of a Heterologous Gene in Escherichia coli in Response to Carbon-Nitrogen Source and C/N Ratio in a Batch Bioreactor, Biotechnol. Prog. 13, 151 (1997) L.J. DeHart, J.T. Winzeler, R.M. Gleason, Rapid Analysis of Monosodium Glutamate (MSG) Utilizing Immobilized Enzyme Sensor Technology, presented at IFT Annual Meeting, July 1993 L.J. DeHart, S. Payne, Immobilized Enzymatic Methods of Sugar Analysis, presented at Conference on Sugar Processing Research, September 1992 R. Engelhardt, M. Vojacek, Determination of Total Choline in Nutritional Products, presented at AOAC Regional Meeting, Central Section, November 1996 R. Engelhardt, J.T. Winzeler, Automated and Rapid Determination of Choline in Nutritional Products Employing Immobilized Choline Oxidase and Electrochemical Detection of Hydrogen Peroxide, presented at IFT Annual Meeting, July 1993 R. Fike, J. Kubiak, P. Price, D. Jayme, Feeding Strategies for Enhanced Hybridoma Productivity: Automated Concentrate Supplementation, BioPharm 6, 49 (1993) G.L. Kleman, J.J. Chalmers, G.W. Luli, W.R. Strohl, A Predictive and Feedback Control Algorithm Maintains a Constant Glucose Concentration in Fed-Batch Fermentations, Applied and Environmental Microbiology 57, 910 (1991) K.B. Konstantinov, Y-S. Tsai, D. Moles, R. Matanguihan, An Aseptic Technique for On-Line Monitoring of the Glucose and Lactate Concentrations in Long-Term Perfusion Cultures, presented at the 209th American Chemical Society National Meeting and Exposition, April 1995 K.B. Konstantinov, Y-S. Tsai, D. Moles, R. Matanguihan, Control of Long-Term Perfusion Chinese Hamster Ovary Cell Culture by Glucose Auxostat, Biotechnol. Prog. 12, 100 (1996) K.B. Konstantinov, Y-S. Tsai, D. Moles, R. Matanguihan, Control of Long-Term Cho Cell Culture by Glucose Auxostat, presented at the 209th American Chemical Society National Meeting and Exposition, April 1995 M. Linz, A-P. Zeng, R. Wagner, W-D. Deckwer, Stoichiometry, Kinetics, and Regulation of Glucose and Amino Acid Metabolism of a Recombinant BHK Cell Line in Batch and Continuous Cultures, Biotechnol. Prog. 13, 453 (1997) G.W. Luli, W.R. Strohl, Comparison of Growth, Acetate Production, and Acetate Inhibition of Escherichia coli Strains in Batch and Fed-Batch Fermentations, Applied and Environmental Microbiology 56, 1004 (1990) M.B. Madaras, R.B. Spokane, J.M. Johnson, J.R. Woodward, Glutamine Biosensors for Biotechnology Applications, with Suppression of the Endogenous Glutamate Signal, Analytical Chemistry 69, 3674 (1997) J.T. Winzeler, Current Trends in Enzyme Sensors, PI Quality, July/August 1993 J.T. Winzeler, D. Moles, Monitoring and Control of Nutrients and Byproducts, Biomedical Products, May 1997 M. Wnukowski, D. Angone, C. Chi. Chou, Application of a Biosensor- Based Instrument for the Monitoring of Sucrose Loss in Refinery Waste Water, presented at Conference on Sugar Processing Research, September 1992 Z. Zhang, J.M. Scharer, M. Moo- Young, Mathematical Model for Aerobic Culture of a Recombinant Yeast, Bioprocess Engineering 17, 235 (1997) 11

12 12 Yellow Springs, Ohio USA Printed in USA 198 B41

YSI Biochemistry Analyzer. Speed and Accuracy Made Simple

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