Abstract

Chronic kidney disease (CKD) is associated with elevated creatinine levels due to impaired kidney function. Conventional creatinine determination is generally performed using blood serum or plasma, requiring invasive sample collection and laboratory-based instrumentation. Although microfluidic paper-based analytical devices (µPADs) have been widely investigated for point-of-care analysis, many reported designs require flow channels, multilayer structures, or complex fabrication procedures. In this study, a spot-based laser-printed microfluidic paper-based analytical device (LP-µPAD) was fabricated on Whatman Grade 4 filter paper for colorimetric creatinine detection. Hydrophobic barriers and hydrophilic detection zones were produced by laser printing followed by thermal treatment. Creatinine was detected using the Jaffe reaction, and the colour formation resulted from the Janovsky complex, which was further analyzed from smartphone images using ImageJ software. The LP-µPAD exhibited a linear response over the concentration range of 0-20 mg/dL, with a limit of quantification of 0.49 mg/dL and limit of detection of 0.15 mg/dL. The assay showed good precision with an RSD of less than 10%, satisfactory selectivity against representative interfering species, and acceptable storage stability under refrigerated conditions. Bland-Altman analysis showed good agreement between the LP-µPAD and the reference UV-Vis method. Recovery values of 99.0-107.5% for artificial saliva and 99.13-112.0% for artificial urine demonstrated satisfactory analytical performance. The proposed spot-based LP-µPAD provides a simple approach for creatinine determination in simulated biological samples.

Keywords

Paper-Based Analytical Device, Creatinine, Janovsky Complex, Colorimetric Sensing, Non-Invasive Diagnostics,

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