Glucose determination with enzyme reactors

Matsumoto et al. (1988) coupled enzyme reactors for the elimination of glucose and ascorbic acid with enzyme reactors for the determination of sucrose, glucose and fructose in an FLA manifold. 

The coupled reactions of isomerase, mutarotase and GOD should be applicable to assay fructose, but a sensor using this sequence has not been described to date. An FIA manifold containing an enzyme reactor with glucose isomerase and mutarotase for the determination of fructose has been developed by Olsson (1987). The glucose formed was measured spectrophotometrically. 

J. Ruz, A. Rios, M.D. Luque de Castro, M. Valcarcel, Determination of glucose in alcoholic beverages by flow injection with two internally coupled injection valves and an enzyme reactor, Anal. Chim. Acta 211 (1988) 281. 

A second example is the determination of glucose, free cholesterol, creatinine, and lactic acid (figure 21.8). This method uses on-line immobilized enzyme reactors followed by chemiluminescence detection of the hydrogen peroxide enzyme reaction product reaction with luminol. Detection limits of 0.1 mg dl were observed. 

Murakami Y, Takeuchi T, Yokoyama K, Tamiya E, Karube I and Suda M 1993 Integration of enzyme-immobilized column with electrochemical flow cell using micromachining techniques for a glucose detection system Anal. Chem. 65 2731-5 Olson B, Stalbom B and Johansson G 1986 Determination of sucrose in the presence of glucose in a flow Injection system with immobilized multi-enzyme reactors Anal. Chim. Acta 179 203-8... 

T. Yao and Y. Kobayashi, Amperometric Determination of Glucose, Galactose, Free Cholesterol and Choline in Serums by Flow Injection Analysis with Immobilized Enzyme Reactors [in Japanese]. Bunseki Kagaku, 32 (1983) 253. 

B. Olsson, B. Stalbom, and G. Johansson, Determination of Sucrose in the Presence of Glucose in a Flow-Injection System with Immobilized Multi-Enzyme Reactors. Anal. Chim. Acta, 179 (1986) 203. 

Matsumoto, K., Kamikado, H., Matsubara, H., Osajima, Y., Simultaneous Determination of Glucose, Fructose, and Sucrose in Mixtures by Amperometric Flow Injection Analysis with Immobilized Enzyme Reactors , Anal. Chem. 60 (1988) 147-151. 

Ruz, J., Luque de Castro, M. D., Valcarcel, M., Determination of Glucose in Alcoholic Beverages by Flow Injection with IWo Internally Coupled Injection Valves and an Enzyme Reactor , Anal Chim. Acta 211 (1988) 281-285. 

Immobilized enzymes are used in the industrial production of many compounds. They are used in the manufacture of amino acids, sugar products such as glucose and high fructose com syrup, and many types of pharmaceutical compounds including antibiotics such as penicillin (1-4). The enzyme is used in these cases for enantioselective synthesis, to produce the desired enantiomer or isomer. Enzymes are currently being explored for their use in the manufacture of biofuels (5). Enzymes are used in analysis in both free and immobilized states. In both states, they are able to determine, with great sensitivity, the concentration of substrates in complex matrices. Immobilized enzymes can be deployed in several different form factors, such as reactor tubes, microwell plates, and as enzyme electrodes in biosensor devices. 

Matsumoto, K., H. Kamikado, H. Matsubara, and Y. Osajima. 1988. Simultaneous determination of glucose, fructose, and sucrose in mixtures by amperometric flow injection analysis with immobilized enzyme reactors. Anal. Chem. 60 147-151. 

Figure 5 Five-channel enzyme sensor for the simultaneous determination of glucose, lactate, glutamate, glutamine, and ammonium. MFM, microfiltration module WV, valves P, pumps DC, dialysis cell B, blank reactors MC, reactor D, biosensor flow cell. (Adapted with permission from Ref. 34.)...

Enzyme Assay Procedure. The catalytic potency of the immobilized g-galactosidase was determined in a plug flow reactor ( 9). Glucose liberated by the catalytic activity of 3-galactosi-dase on lactose was determined by the glucose oxidase-chromogen method (21 ) with some modifications. 

Table 4 shows the calculated values of conversion, productivity, and yield for each residence time tested the productivity for the maximum conversion reached in the continuous SSF (97%) was 5.9 g/l/h, with yield of 81%, for a residence time of 11.1 h, calculated using the average values of the four determined TRS and ethanol concentrations for this residence time. In the discontinuous SSF reactor, for the same enzyme and yeast concentrations, the results were total conversion of TRS in glucose in 6 h and 99.8% of glucose/ethanol conversion in 9 h, with high concentrations of free glucose in the medium. 

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