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Fast, Accurate and Reproducible Method for Simultaneous Quantitation of Pyridoxal 5’-phosphate and Pyridoxic Acid in Human Plasma by HPLC with Fluorescent Detection

Phenomenex Inc
17 Dec 2014

Fluorescence detection has been used nearly exclusively by a majority of reference laboratories in HPLC analysis of vitamin B6 due to its selectivity and high sensitivity. However, the published derivatization procedures suffer from tedious sample preparation and short HPLC column lifetimes. This study aimed to simplify the derivatization and sample preparation steps when analyzing PLP and pyridoxic acid (PA) by HPLC-FLD.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Clinical ChemistryBiochemistry (or clinical chemistry) involves the analysis of bodily fluids using chemical tests. Techniques used include HPLC, chromatography, spectroscopy, mass spectrometry, immunochemical, electrophoresis, turbidometric / spectrophotometric assay, MRI and ISE analysis. Tests are often carried out on plasma or serum but urine (urinalysis) and fecal specimens are also processed.HPLC Method DevelopmentIt is important to develop and validate HPLC methods in order to identify, quantify or purify compounds of interest in the most efficient way. Effective method development ensures optimization, quality control and cost effectiveness.Molecular DiagnosticsMolecular diagnostics use an individual’s genetic code and gene expression to diagnose and monitor diseases. The technique is used increasingly in the field of infectious diseases and oncology, as well as areas such as coagulation, HLA typing and pharmacogenomics. Molecular diagnostics plays a pivotal role in personalized medicine.

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Fast, Accurate and Reproducible Method for Simultaneous Quantitation of Pyridoxal 5’-phosphate and Pyridoxic Acid in Human Plasma by HPLC with Fluorescent Detection