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Analytical Quality by Design-Based Reverse-Phase High-Performance Liquid Chromatography Method Development for
Sakshi B Patil1, Shubhangi B Sutar1, Rachana Bhimanwar2,3
1Ashokrao Mane College of Pharmacy, Peth Vadgaon, India.
Biomedical Chromatography : BMC
|July 25, 2026
Summary
Analytical Quality by Design (AQbD) improves biomarker analysis using RP-HPLC. This systematic approach enhances method robustness and reproducibility, addressing limitations of traditional trial-and-error techniques for better disease diagnosis and monitoring.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Pharmacology
Background:
- Biomarkers are vital for disease diagnosis and treatment monitoring.
- Reverse-phase high-performance liquid chromatography (RP-HPLC) is a common technique for biomarker analysis.
- Conventional RP-HPLC method development relies on inefficient trial-and-error strategies, leading to poor reproducibility and robustness.
Purpose of the Study:
- To critically review existing RP-HPLC methods for biomarker analysis through the lens of Analytical Quality by Design (AQbD) principles.
- To identify limitations in current methods and propose AQbD-based strategies for improvement.
- To discuss emerging trends in sustainable analytical method development.
Main Methods:
- Review of published RP-HPLC methods for biomarker analysis.
- Evaluation of AQbD elements: Analytical Target Profile (ATP), Critical Quality Attributes (CQAs), Critical Method Parameters (CMPs), risk assessment, Design of Experiments (DoE), and Method Operable Design Region (MODR).
- Analysis of common method limitations like coelution and sensitivity.
Main Results:
- Most reviewed studies exhibit incomplete AQbD implementation, lacking multivariate optimization and design space establishment.
- Identified common issues include coelution, insufficient sensitivity, and inadequate method robustness.
- AQbD principles offer effective strategies to overcome these analytical challenges.
Conclusions:
- AQbD provides a systematic framework to enhance the reliability and robustness of RP-HPLC methods for biomarker analysis.
- Addressing limitations through AQbD leads to improved analytical performance for diagnostics and therapeutics.
- Integration of machine learning and Green Analytical Chemistry principles can further advance sustainable biomarker analysis.
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