Simultaneous Detection of Glucose and Acetoacetate in Artificial Urine Samples Using 3D-Connector Microfluidic Paper Based Analytical Devices

Diabetic patients sometimes experience both hyperglycemia and hyperketonemia simultaneously. This condition requires proper management of glucose and ketone body biomarkers. The fluctuating intensity of these biomarkers necessitates regular monitoring. This study aims to develop concept non-enzymatic measurement method by integrating µPADs (microfluidic paper-based analytical devices) for detecting glucose and acetoacetate in artificial urine samples. The method uses silver nanoparticle formation for glucose detection and Schiff base reaction as well as the Rothera test for acetoacetate detection. Optimal conditions found include the glucose detection zone with volume ratio of AgNO3 500 mM : starch 3% (w/v) (1 : 1), and the acetoacetate detection zone with glycine 900 mM in phosphate buffer pH 9.4. The artificial urine sample combination consists of glucose, acetoacetate, and acetone with volume ratio of (1 : 1 : 1). The 3D-Connector for glucose uses NaOH 10 M : starch 3% (w/v) with volume ratio of (1 : 3), while for acetoacetate, sodium nitroprusside 15% (w/w) in DMF 5% (v/v). Validity for glucose measurement shows linearity (R² = 0.9664), precision (RSD = 4.56%), accuracy (88.75 – 99.62%), LOD (1.61 mM), and LOQ (5.37 mM). Conversely, acetoacetate measurement shows linearity (R² = 0.9636), precision (RSD = 1.24%), accuracy (99.44 – 99.73%), LOD (1.41 mM), and LOQ (4.69 mM).