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Routine Isotope-Dilution Liquid Chromatography-Tandem Mass Spectrometry Assay for Simultaneous Measurement of the 25-Hydroxy Metabolites of Vitamins D^sub 2^ and D^sub 3^
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Clinical Chemistry
- Article date:
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September 1, 2005
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Copyright informationCopyright American Association for Clinical Chemistry Sep 2005. Provided by ProQuest LLC. (Hide copyright information)
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Background: Measurement of 25-hydroxyvitamin D^sub 2^ and D^sub 3^ (25-OH D^sub 2^ and D^sub 3^) is essential for investigating vitamin D deficiency. Competitive binding techniques are unable to distinguish between the 2 metabolites and suffer from interference from other hydroxy metabolites of vitamin D.
Methods: We used isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) for routine determination of 25-OH D^sub 2^ and D^sub 3^ with a stable-isotope-labeled internal standard (IS). Serum samples (100 μL) were denatured with methanol-propanol containing IS, vortex-mixed, extracted into hexane, and dried under nitrogen. The reconstituted extract was ...
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