Development of the simultaneous colorimetric enzymatic detection of sucrose, fructose and glucose using a microfluidic paper-based analytical device.

Herein, we propose the first fabrication of a microfluidic paper-based analytical device (μPAD) as a simple, rapid and selective method for the simultaneous detection of sucrose, fructose and glucose for quality monitoring in the sugar industry. Multiplex detection of the three analytes was achieved by three detection zones designed on a single device. Each detection zone on the μPAD was modified with enzymes, chromogenic reagents and specific substrates. The enzymatic reactions between the specific enzymes, chromogenic reagents and their corresponding substrates yielded different colored products: brown for sucrose, blue for fructose and pink for glucose. All the reaction conditions were evaluated. The limits of detection were determined to be 0.9 mM, 0.8 mM and 0.4 mM for sucrose, fructose and glucose, respectively. The proposed μPAD successfully monitored sucrose, fructose and glucose within 11 min using a signal device and obtained a spike recovery of 100-103% for sucrose, 98-103% for fructose, and 100-102% for glucose. The μPAD was applied to detect sucrose, fructose and glucose in real samples, and the results were in good agreement with those obtained using high-performance liquid chromatography (HPLC). The proposed μPAD provides several advantages, including ease of use, fast results and low cost. Therefore, the developed μPAD is suitable for application in the sugar industry for monitoring sucrose, fructose and glucose in parallel.