Analytical Method Validation for the Determination of Ninhydrin Positive Substances in Amino Acids by High Performance Liquid Chromatography

Abstract

The aim of the work is focused on the Validation of an Analytical method for the determination of Ninhydrin Positive substances (NPS) in Amino acids using HPLC Technique in a single method. Since, most of the amino acids are widely used in the determination of Renal and Nutrition drug products.The Chromatographic separation was performed using Sodium Amino Acid Analysis Cation Exchange column using Sodium Eluent Na 315, Sodium eluent Na 425 and Sodium Eluent Na 640 as Mobile phase, performed by Gradient program with detection of wavelength 570 nm using flow rate as 0.4 mL/min. The method has been developed and validated using post column derivatization technique (HPLC/Pinnacle PCX).The Method Validation Parameters/Performance Characteristics like Specificity, Method Precision, Linearity and Range, LOD, LOQ, Ruggedness, Solution stability, Robustness was carried out and all the amino acids were successfully met the acceptance criteria and the method has been validated as per ICH Q2R1 Guideline.The method has been successfully validated for its intended purpose.