Sequential enzymatic quantification of two sugars in a single microchannel

Abstract

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.

Microfluidic enzymatic biosensing systems: A review

“…Li et al, 2010a), cholesterol (Aravamudhan et al, 2007;Wisitsoraat et al, 2010) and renal markers such as urea Malecha et al, 2009), creatinine (Satoh et al, 2008;Suzuki and Matsugi, 2005), alanine aminotransferase (Song et al, 2009) or glutamic-oxaloacetic transaminase . In food and environmental monitoring, enzymatic biosensing systems have been developed for the detection of fructose (Atalay et al, 2012), lactose in milk (Fornera et al, 2012;Ito et al, 2011), pesticide residues in vegetable and soil samples (Duford et al, 2013) or hypoxanthine to determine the freshness of fish meat (Dolmaci et al, 2012;Lawal and Adeloju, 2010). Another interesting application field is the monitoring of nutrients and metabolic products in cell cultures and biotechnological processes.…”

“…10C) (Songjaroen et al, 2009) which create turbulences can be integrated into the channels. In systems dedicated to electrophoretic measurements, also the different electrophoretic mobility of species and sequential injection schemes can be employed for mixing (Atalay et al, 2012;Chun-Mao et al, 2007). Substances with higher mobility in the electrophoresis channel overtake those with lower mobility and mixing takes place (Fig.…”