Method Validation and Uncertainty Estimation for Total Phosphorus Determination in Animal Feed Using UV-Vis Spectrophotometer

Abstract

Objective: The study presents the validation protocol for the determination of total phosphorus content at parts per million (ppm) (μg/L) levels in animal feed by a UV-Vis spectrophotometer. Methods: The measured absorbance of solutions against the blank solution was at 400 nm with the spectrophotometer. A combined ammonium heptamolybdate tetrahydrate and ammonium monovanadate blue solution was used as a coloring reagent for detection. This method was validated by evaluation of statistical parameters such as linearity, sensitivity, limits of detection (LOD) and quantification (LOQ), precision, accuracy, and measurement uncertainty using a matrix blank (MB) against the phosphorus standard. Results: The Instrumental Detection Limit was 0.066 ppm and the Instrumental Quantification Limit was 0.22 ppm, respectively, while the phosphorus recovery and repeatability percent were 101.15% and 0.11%, respectively. However, the linearity of this method was 0.1 to 30 ppm. The measurement uncertainty of this method was 2.82%, following Commission Regulation (EC). Conclusion: The estimated parameters in the validation protocol, were found to meet the imposed performance criteria, and the procedure was validated for the intended use.