HPLC and LCMS – A review and a recent update

Abstract

Chromatography is defined as a set of techniques which is used for the separation of constituents in a mixture. This technique involves 2 phases stationary and mobile phases. The separation of constituents is based on the difference between partition coefficients of the two phases. The chromatography term is derived from the greek words namely chroma (colour) and graphein (to write). The chromatography is very popular technique and it is mostly used analytically. There are different types of chromatographic techniques namely Paper Chromatography, Gas Chromatography, Liquid Chromatography, Thin Layer Chromatography (TLC), Ion exchange Chromatography and lastly High Performance Liquid Chromatography (HPLC). This review mainly focuses on the HPLC technique its principle, types,  instrumentation and  applications. Liquid  Chromatography/Mass Spectrometry (LC/MS) is  fast developing and it‟s the preferred tool of liquid chromatographers. Liquid chromatography-mass spectrometry (LC- MS/MS) is a technique that uses liquid chromatography (or HPLC) with the mass spectrometry. It is an analytical chemistry technique that combines the physical separation capabilities of liquid chromatography (or HPLC) with the mass analysis capabilities of mass spectrometry. (LC-MS/MS) is commonly used in laboratories for the qualitative and quantitative analysis of drug substances, drug products and biological samples.  It has been persistently used in drug development at many different stages including Metabolic Stability Screening, Metabolite Identification as well as In Vivo Drug Screening, Impurity Identification, Peptide Mapping, Glycoprotein Mapping, Natural Products De replication, Bio-affinity Screening. LC-MS is now successfully applied to routine analysis in many areas, including therapeutic drug monitoring (TDM), clinical and forensic toxicology as well as doping control. This advancement in LCMS was originally and still is fueled by the need for more powerful analytical and bio-analytical techniques that can accurately and precisely discriminate target analytes from high complexity mixtures in a sensitive and selective way. With recent advancement in instrumentation, the use of liquid chromatography (LC) and mass spectrometry (MS) has become a powerful two-dimensional (2D) hyphenated technology.