Glucose biochip: Dual analyte response in connection to two pre-column enzymatic reactions

This article describes a novel 'Lab-on-a-Chip' protocol generating two electrophoretic peaks for a single analyte, based on the coupling of two different pre-column enzymatic reactions of the same substrate followed by electrophoretic separation of the reaction products. Such operation is illustrated for the measurement of glucose in connection to the corresponding glucose oxidase (GOx) and glucose dehydrogenase (GDH) reactions. The pre-column enzymatic reactions generate hydrogen peroxide and NADH species, that are separated (based on their different charges) and detected at the end-column amperometric detector. The peak current ratio can be used for confirming the peak identity, estimating the peak purity, addressing co-migrating interferences, and deviations from linearity. A driving voltage of 2000 V results in peroxide and NADH migration times of 93 and 260 s, respectively. Factors influencing the unique dual glucose response are examined and optimized. The concept can be extended to different target analytes based on the coupling of two pre-column reactions with electrophoretic separation of the reaction products.