Analytical Method Validation for Quantification of Active Pharmaceutical Ingredients (APIs): A Comparative Study of HPLC and UV-Spectrophotometry

Abstract

Correct determination of Active Pharmaceutical Ingredients (APIs) will play a crucial role in the quality, safety, and compliance of pharmaceutical products. The purpose of the study is to provide a comparative assessment of the High-Performance Liquid Chromatography (HPLC) and UV-Spectrophotometry as the methods used in the determination of API using the ICH Q2(R1) criteria of complete analytical method validation. Both methods were developed using methods of optimizing analytical conditions, standard and sample solution preparation, and evaluation of major validation parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD), and limit of quantification (LOQ) and robustness. The results showed that HPLC had a better analytical performance, higher precision, accuracy, lower LOD values, and low LOQ values and higher resistant to slight procedural variations. Although somewhat less accurate and prone to interference, UV-Spectrophotometry was a quick and cost-effective and convenient technique to conduct daily quality control and primary analyses. Comparative analysis revealed that HPLC is more applicable to complex formulations involving a high degree sensitivity and resolution level, whereas UV is useful in resource- restrained environment and simple-to-quantify tasks. Summing up, the paper emphasizes the advantageous features of the two methods and emphasizes the necessity of using the method depending on the analytical needs, the complexity of formulation, and the available laboratory facilities. The findings will be useful in enhanced decision-making in development and validation of pharmaceutical analytical methods.