Quantitative subfemtomole analysis of alpha-tocopherol an deuterated isotopomers in plasma using tabletop GC/MS/MS

C K Van Pelt, P Haggarty, J T Brenna, Paul Haggarty

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    Abstract

    A rapid, high-selectivity method with subfemtomole sensitivity is reported for quantification of alpha-tocopherol in plasma-based gas chromatography/tandem mass spectrometry (GC/MS/MS) using a tabletop quadrupole ion trap mass spectrometer. Sample workup is rapid, consisting of protein precipitation followed by liquid/liquid extraction and O-trimethylsilyl derivatization of alpha-tocopherol (alpha-T-TMS) and an internal standard, 2,2,5,7,8-pentamethyl-6-chromanol (PC-TMS). Rudimentary chromatography was carried out using an 8-m DB-5 capillary column resulting in an analyte retention time of 7.2 min. No interferences from the plasma matrix were observed. The assay has a detection limit of 178 amol (89.6 fg) and a lower limit of quantification of 700 amol (350 fg)of derivatized alpha-tocopherol in diluted plasma; <30 pL of plasma is estimated to yield sufficient alpha-tocopherol for quantitative analysis at typical concentrations found in humans. A calibration curve constructed from National Institute of Standards and Technology serum standards was linear in the working range of 1.9-1073 ng/mL (0.95-0.54 ng). Within- and between-day precision averaged 5.8% and did not exceed 11.3% for three concentrations of quality control (QC) solutions. The overall accuracy for the QC samples was within 7.2%. Storage studies showed that (alpha-T-TMS and PC-TMS are stable under conditions that might be encountered during analyses. In a test study, plasma kinetic curves for alpha-tocopherol-d(6) and alpha-tocopherol-d(3) were obtained for a catheterized pregnant ewe and her fetus who were simultaneously given a bolus injection of alpha-tocopherol-d(6), to the ewe and alpha-tocopherol-d(3) to the fetus. These data show that a tabletop GC ion trap can determine alpha-TT-TMS and its isotopomers quantitatively at high selectivity in a complex matrix.

    Original languageEnglish
    Pages (from-to)4369-4375
    Number of pages7
    JournalAnalytical Chemistry
    Volume70
    Issue number20
    Publication statusPublished - 15 Oct 1998

    Keywords

    • TANDEM-MASS-SPECTROMETRY
    • GAS-CHROMATOGRAPHY
    • OXIDATION-PRODUCTS
    • VITAMIN-E
    • PERFORMANCE
    • DRUG
    • TEBUFELONE
    • RESIDUES
    • URINE
    • LEVEL

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