Chromatography Techniques

Chromatography-HPLC in Bio-Medical research

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HPLC is the most versatile of all chromatography methods but also the most complex. It was first made available in the laboratory during the 1970s and is currently used for the analysis of amino acids, peptides, proteins, carbohydrates, lipids, nucleic acids and related compounds, vitamins, hormones, metabolites, and drugs. HPLC can be coupled to various detectors such as UV, fluorescence or mass spectrometry (LC/MS and LC/MS/MS) and is routinely used for quantitative analysis in biological samples such as blood, urine and other body fluids. HPLC consists of using a liquid mobile phase to pass under high pressure a mixture of analytes extracted from the sample through a column containing the stationary phase. Analyte separation is based on differences in interaction with both the mobile phase and the stationary phase.

HPLC is a proven method for isolating analytes of interest in complex matrices such as biological fluids. Its use in the clinical laboratory has steadily increased over the past decades as its unmatched analytical performance and versatility allows for testing of many different types of clinically relevant analytes. With the recent advances in detection technology such as mass spectrometry and sample preparation techniques such as bio-affinity chromatography and online automation, HPLC--‐based methods will likely remain the gold standard of clinical testing for many of the current but also future biomarkers and therapeutic drugs.