Simultaneous and sensitive detection of multiple small biological molecules by microfluidic paper-based analytical...

In this work, ZnO nanorods (ZnO NRs) with different sizes were hydrothermally grown on the surface of Whatman filter paper for the fabrication of a microfluidic paper-based device (μPAD) for the simultaneous detection of glucose and uric acid. As dual enzymatic reaction was employed for the colorimetric detection in this μPAD, the presence of ZnO NRs promoted the enzyme immobilization thus significantly enhancing the colorimetric signal. The coffee ring effect was effectively conquered by the uniform distribution of ZnO NR as well as a specialized double-layered μPAD design. Meanwhile, two color indicators with distinct colors were used to provide complementary results to better quantify the concentration of the analytes by naked eye. As a result, two linear calibration curves were obtained for the detection of glucose (0.01-10 mmol L^-1) and uric acid (0.01-5 mmol L^-1), along with a LOD of 3 μmol L^-1 for glucose and 4 μmol L^-1 for uric acid, respectively. The practical usefulness of the proposed μPAD was further validated by the simultaneous analysis of glucose and uric acid in serum samples and urine samples.