Biochemical Applications of Chromatography-HPLC

HPLC is a very common method for metabolomics analysis. With the invention of electrospray ionization, HPLC is coupled to mass spectroscopy. HPLC has lower chromatographic resolution, requires no derivation for polar molecules and separates molecules in the liquid phase. HPLC has the advantage of much wider range of analytes measurements with a higher sensitivity than gas chromatographic methods. Relevant to proteomics, due to the complex structure and nature of proteins, instrumentation and methods development for sample clean-up, pre-concentration, fractionation, chromatographic separation and detection becomes an immediate requirement for the identification of peptides and proteins. HPLC is considered as most reliable and most sensitive technique in genomics used to determine DNA methylation. The nucleosides and nucleotides of DNA are separated and quantified by HPLC-UV method. HPLC finds applications in glycomics and lipidomics where glycan part is cleaved either enzymatically or chemically from the target and subjected to analysis. 

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