Simultaneous microchip enzymatic measurements of blood lactate and glucose.
A miniaturized capillary electrophoretic (CE) microchip device for the simultaneous measurements of lactate and glucose is described. The new microchip bioassay protocol integrates an electrophoretic separation of lactate and glucose, post-column enzymatic reactions of these metabolites with their respective oxidase enzymes, and an amperometric (anodic) detection of enzymatically-liberated hydrogen peroxide at a gold-coated thick-film carbon detector. Factors influencing the response have been examined and optimized, and the analytical performance has been characterized. Applicability of the microchip assay to clinical samples, such as serum and blood, is demonstrated. The microchip protocol obviates cross enzymatic reactions and interferences from major oxidizable constituents common to dual glucose-lactate enzyme electrodes. Such ability to rapidly separate and quantitate lactate and glucose on a small microchip platform should find important clinical and biotechnological applications.
Wang
Joseph
J
Departments of Chemical Engineering and Chemistry and Biochemistry, Biodesign Institute, Arizona State University, Tempe, AZ 85287-5001, USA. joseph.wang@asu.edu <joseph.wang@asu.edu>
Chatrathi
Madhu Prakash
MP
Collins
Greg E
GE
D000328:Adult; D001234:Aspergillus niger / Q000201:enzymology; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods; D001786:Blood Glucose / Q000737:chemistry*; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D056656:Lab-On-A-Chip Devices; D007773:Lactates / Q000097:blood* / Q000737:chemistry; D008297:Male; D046208:Microchip Analytical Procedures / Q000379:methods; D010376:Pediococcus / Q000201:enzymology; D013696:Temperature
D001786:Blood Glucose; D007773:Lactates; D006861:Hydrogen Peroxide