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February 2, 20260 citationsOpen Access

Method Development and Validation by HPLC (A Review)

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*W*1Dr. Ananthakrishnan, 2Dr. Jeyavelraman Kiruthika, 3Mr. P. Sriram, 4Mr. A. Raj Kumar, 5Mr. S. Jeevan Nithish, 6Gopal Dilip Wagh

Key Points

  • This review aims to summarize the essential processes involved in developing and validating HPLC methods for drug analysis.
  • Evaluating chromatographic factors such as sample preparation and mobile phase selection.
  • Focusing on the choice of column and detector type for method optimization.
  • Discussing compliance with Good Manufacturing Practice (GMP) during validation processes.
  • HPLC is identified as a highly efficient method for drug detection and quantification.
  • The review outlines the significance of method validation for ensuring accuracy and precision in pharmaceutical applications.
  • Highlights the necessity of a comprehensive validation policy for adherence to regulatory standards.

Abstract

High-Performance Liquid Chromatography (HPLC) is one of the most widely used analytical techniques for the detection, separation, and quantification of drugs. During method optimization, various chromatographic factors are evaluated, such as sample preparation, selection of the mobile phase, choice of column, and type of detector. This article focuses on the processes involved in HPLC method development, optimization, and validation. Due to its benefits, including high speed, specificity, accuracy, precision, and suitability for automation, HPLC is extensively used for the analysis of drugs in multicomponent pharmaceutical dosage forms. The development and validation of HPLC methods play a vital role in new drug discovery, formulation development, manufacturing, and in both human and veterinary studies. Analytical method validation is an essential requirement during drug development and production to confirm that the method is appropriate for its intended application. To comply with Good Manufacturing Practice (GMP) standards, pharmaceutical industries must establish a comprehensive validation policy that clearly describes validation procedures. This article mainly emphasizes the optimization of HPLC operating conditions.

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*1Dr. Ananthakrishnan, 2Dr. Jeyavelraman Kiruthika, 3Mr. P. Sriram, 4Mr. A. Raj Kumar, 5Mr. S. Jeevan Nithish, 6Gopal Dilip Wagh (2026) studied this question.

synapsesocial.com/papers/6980fe7cc1c9540dea8108fehttps://doi.org/10.5281/zenodo.18439970
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  5. 5ULTRA-PERFORMANCE LIQUID CHROMATOGRAPHY (UPLC) BASED ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR PHARMACEUTICAL DOSAGE FORMS: A COMPREHENSIVE REVIEW2026