Review on Advanced RP-HPLC Method Development for Multicomponent Analysis: Techniques, Applications, and Validation Protocols

RP-HPLC or Reverse Phase High-Performance Liquid Chromatography is easy to use by highly trained specialists, with applications in pharmaceutical analysis, food technology, and even environmental science. It is a technology that combines two techniques, high chromatography and special compound separation. In this review, I will discuss stationary and mobile phase matters, optimization parameters for RP-HPLC systems, multi component multiseries advanced detectors, and the challenges of multi component detection using modern detectors and zeotropic chromatographic techniques. Central to new technology is improvement in productivity and efficiency and decreases in the amount of energy used and the effect on the environment with increased sustainability, both aspects important in contemporary medicine and science. The approach ‘Quality by Design’ employs the development of green mobile phases along with novel stationary and hybrid systems. Sharper analysis, stronger sensitivity, and increasing the number of separable complex components in one CRAM allow for more versatile methods. Different practices of RP-HPLC in pharmaceutical, environmental, food and beverage industries, as well as biological and biomedical research, highlight the product’s quality, safety, and efficacy. This review analysis highlights the significance of method validation, methods to solve specific problems, and best practices for positive outcomes for results reliability and reproduction. Potential future directions and advancements in RP-HPLC, which include miniaturization, automation, and sustainability are also covered, showing the prospects of more compact, efficient, and eco-friendly systems for chromatography.