Creatinine, an important biomarker for renal functions, is often conventionally estimated using gold standard Jaffe reaction from blood; which involves the usage of a spectrophotometric measurement; thus, restricts it’s utilitarian scopes in point-of-care settings. Here, we report the development of a single step quantification of creatinine from whole blood using a paper-based microfluidic device. Our platform uses Whatman filter paper integrated with LF1 membrane; allows on-chip separation of blood plasma through LF1 membrane, while Whatman component of the device contains the embedded reagents for Jaffé reaction. Combination of two different grades of paper allows a single step quantification of creatinine as the separated blood plasma traverses to the reaction zone through capillary imbibition. Colorimetric readouts were quantified using an RGB sensor instead of a smartphone which is highly platform dependent rendering relatively higher cost rather than other components of point-of care (POC) devices. Our sensor has been integrated within a 3D box; thereby virtually making the detection instrument free and perfectly suited for POC settings. The limit of detection (LOD) of our device is 0.219 mg/dL, which falls within the lower range of physiological values. The coefficient of determination (R²) for linearity and median accuracy have been found as 0.978 and 94.047%, respectively. The relative standard deviation (RSD) for precision measurements has remained below 5% for the developed protocol. Furthermore, we have validated the performance of our device with 35 clinical samples in laboratory settings against the gold standard measurements. Our Bland Altman plot as well as t-test and Chi-square test clearly confirms the validity of our device within 95% confidence interval.

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