This paper describes the modification of microfluidic paper-based analytical devices (μPADs) with chitosan to improve the analytical performance of colorimetric measurements associated with enzymatic bioassays. Chitosan is a natural biopolymer extensively used to modify biosensing surfaces due to its capability of providing a suitable microenvironment for the direct electron transfer between an enzyme and a reactive surface. This hypothesis was investigated using glucose and uric acid (UA) colorimetric assays as model systems. The best colorimetric sensitivity for glucose and UA was achieved using a chromogenic solution composed of 4-aminoantipyrine and sodium 3,5-dichloro-2-hydroxy-benzenesulfonate (4-AAP/DHBS), which provided a linear response for a concentration range between 0.1 and 1.0 mM. Glucose and UA were successfully determined in artificial serum samples with accuracies between 87 and 114%. The limits of detection (LODs) found for glucose and UA assays were 23 and 37 μM, respectively. The enhanced analytical performance of chitosan-modified μPADs allowed the colorimetric detection of glucose in tear samples from four nondiabetic patients. The achieved concentration levels ranged from 130 to 380 μM. The modified μPADs offered analytical reliability and accuracy as well as no statistical difference from the values achieved through a reference method. Based on the presented results, the proposed μPAD can be a powerful alternative tool for non-invasive glucose analysis.

This paper describes a paper-based colorimetric biosensor for measuring glucose concentration levels in human tear samples. Colorimetric biosensors were wax printed on paper platforms and modified with chitosan previously prepared in acetic acid. The proposed device was explored to measure the glucose levels in human tear samples using 3,3′,5,5′-tetramethylbenzydine (TMB) as the chromogenic reagent. The paper-based colorimetric biosensor exhibited a linear behavior for the glucose concentration range between 0.1 and 1.0 mM. The achieved analytical sensitivity and limit of detection (LOD) were 84 AU/mM and 50 µM, respectively. Moreover, the device provided analytical reliability and no statistical difference when compared to the data recorded with a commercial glucometer. The proof-of-concept of our device was successfully demonstrated by measuring the glucose levels in six tear samples from nondiabetic subjects. In general, the results showed that the colorimetric biosensor has noticeable potential to be used as a powerful tool for tear glucose monitoring, since this fluid offers lower potential interferences, non-invasive sample collection and is pain-free. Furthermore, the proposed device could facilitate the treatment of diabetic patients who need constant control of glucose levels and cannot tolerate multiple finger sticks per day.

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