Sequential enzymatic quantification of two sugars in a single microchannel

This paper presents the implementation of a multiple analyte enzyme assay, based on the sequential injection of the different enzyme solutions, in an electrokinetic driven microfluidic chip. The assay methodology for the simultaneous quantification of d -glucose and d -fructose was reported in previous publications but here the real integration of both enzyme assays was achieved. When assays were executed separately, good reproducibility was observed with average CV values of 5.2% and 4.5% for the d -glucose and d -fructose assay, respectively. Next, the assays for the quantification of d -glucose and d -fructose were integrated simultaneously on chip, where each assay was executed consecutively in the same microreactor by applying a specific sequence of potentials at the reservoirs. This article proves the integration of a sequential based quantification approach in continuous microfluidic chips with electrokinetic actuation.