Manjeet Baliyan
Abstract
This paper provides a comprehensive overview of the evolution and applications of
chromatography, with a focus on High-Pressure Liquid Chromatography (HPLC) in
pharmaceutical analysis. Chromatography, initially discovered by Russian botanist Mikhail
Tswett in the 19th century, has seen significant advancements since its inception, leading to
various chromatography types and applications.
The narrative traces the historical development, highlighting Tswett’s pioneering work in plant
color categorization and the emergence of true chromatography. The 1940s witnessed the
evolution of chromatography types, while the 1960s marked a pivotal moment with Csaba
Harwath’s creation of the first liquid chromatographic equipment. In the late 1960s, Kirkland’s
incorporation of a flow cell for a UV detector played a crucial role in advancing High-
Performance Liquid Chromatography (HPLC).
The focus then shifts to HPLC, a powerful separation method extensively used in pharmaceutical
analysis. The paper details the principles of chromatography, advantages of HPLC, and the
various types of separation it offers, including Normal Phase and Reverse Phase
Chromatography. The instrumentation of HPLC is explored, and the crucial role of buffers in
HPLC separations is emphasized.
The selection of solvents, columns, and mobile phases in HPLC is discussed in detail, addressing
the importance of solubility, absorbance, and water in mobile phases. The paper also delves into
degassing techniques and highlights the significance of system suitability parameters for method
validation.
The latter part of the paper provides an in-depth exploration of analytical method development
using HPLC, covering key considerations such as chromatography mode, detector selection, and
optimization of mobile phases. The validation parameters for HPLC methods, including system
suitability, efficiency, retention time, and resolution factor, are thoroughly explained in
accordance with ICH guidelines.
Keywords: Chromatography, High-Pressure Liquid Chromatography (HPLC), Pharmaceutical Analysis, Analytical Method Development, Gas Chromatography (GC)
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| Volume | |
| Issue | |
| Received | January 12, 2024 |
| Accepted | January 19, 2024 |
| Published |
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