Abstract
This paper presents an enzyme-based method for the determination of urea in aqueous samples using crude urease from Citrullus lanatus seeds. The method is based on the enzymatic hydrolysis of Urea into ammonia (NH4) and carbon dioxide (CO2) by urease. Ammonia formation is determined spectrophotometrically after nesslerization and then quantitatively related to urea concentration via a calibration curve. Reaction conditions for the assay were investigated and optimized. Optimum temperature was found to be 50 oC, optimum pH was 8.0 and optimum reaction time was 10 minutes. The Calibration curve for urea determination was linear (R2 =0.987) within the concentration range of 0.5 – 50 mg/L. Limits of detection and quantification (LOD and LOQ) for the method were 0.08 and 0.13 mg/L respectively. Application of the method to urea determination in surface water samples revealed good precision (1.45 – 3.28 % RSD) and accuracy (84 – 92% recovery). The presented method is simple and has the potential to be a practical tool for monitoring of urea in environmental samples.
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