Continuous flow microfluidic system with magnetic nanoparticles for the spectrophotometric quantification of urea in urine and plasma samples.

Urea, synthesized exclusively in the liver, is primarily transported through the bloodstream to the kidneys, where it is excreted in urine, accounting for 80-90% of nitrogen excretion in humans. Elevated blood urea levels, indicative of kidney dysfunction, make it a crucial biomarker for assessing renal function. Previous studies on urea detection using microdevices have largely focused on conductometric methods. In this study, we demonstrated the application of a continuous flow miniaturized system for rapid spectrophotometric urea quantification using polydimethylsiloxane (PDMS) microdevices. The microdevice featured two distinct zones: an enzymatic reaction zone, where urease-conjugated magnetic nanoparticles were immobilized, and a detection zone, where reagents were incorporated to produce a colored reaction product a modified Berthelot reaction. Integrating magnetic nanoparticles as a solid support for the enzyme enabled the reuse of PDMS microdevices without compromising the analytical signal. Spectrophotometric detection was performed in an additional microdevice acting as a microflow cell coupled with optical fibers. A calibration curve was constructed using urea standards diluted in phosphate buffer solution (PBS), yielding a linear range of 0.12-3.00 mg dL. The method demonstrated detection and quantification limits of 0.04 mg dL and 0.12 mg dL, respectively. Precision and accuracy assessments yielded a repeatability of 0.90% and intermediate precision of 4.52%, with recovery rates near 100%. The method was applied to plasma and urine samples, showing urea concentrations within normal physiological ranges and an analysis throughput of 36 measurements per hour.