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Rapid Separation of 25-OH-vitamin D3 and 3-epi-25-OH-vitamin D3 in Human Serum Under RP-LC Conditions and Tandem Mass Spectrometry Detection
26 Feb 2013Faulty vitamin D metabolism in children less than 12 months of age can lead to formation of the inactive 3-epi-25 monohydroxy form. The resolution of 3-epimer from the active monohydroxy form by tandem mass spectrometry is not possible due to the mostly identical fragmentation pattern of the two species. As a result, the two isomers should be separated chromatographically. This application note describes an assay that utilizes the Kinetex 2.6 μm PFP column that can accurately quantitate 25-OH-D2, 25-OH-D3, and the 25-OHD3 epimer with a total analysis time of less than 5 minutes.
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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.Vitamin D DeficiencyVitamin DChromatography Method DevelopmentChromatography method development is the process of designing and optimizing chromatographic techniques to separate and analyze complex mixtures. This includes selecting appropriate stationary and mobile phases, determining flow rates, and optimizing detection methods. Successful method development is essential for high-quality results in fields like pharmaceuticals, food safety, and environmental analysis. Explore chromatography method development tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.