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

    Research output: Contribution to journalArticle

    33 Citations (Scopus)

    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

    Cite this

    Quantitative subfemtomole analysis of alpha-tocopherol an deuterated isotopomers in plasma using tabletop GC/MS/MS. / Van Pelt, C K ; Haggarty, P ; Brenna, J T ; Haggarty, Paul.

    In: Analytical Chemistry, Vol. 70, No. 20, 15.10.1998, p. 4369-4375.

    Research output: Contribution to journalArticle

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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.",
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    T1 - Quantitative subfemtomole analysis of alpha-tocopherol an deuterated isotopomers in plasma using tabletop GC/MS/MS

    AU - Van Pelt, C K

    AU - Haggarty, P

    AU - Brenna, J T

    AU - Haggarty, Paul

    PY - 1998/10/15

    Y1 - 1998/10/15

    N2 - 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.

    AB - 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.

    KW - TANDEM-MASS-SPECTROMETRY

    KW - GAS-CHROMATOGRAPHY

    KW - OXIDATION-PRODUCTS

    KW - VITAMIN-E

    KW - PERFORMANCE

    KW - DRUG

    KW - TEBUFELONE

    KW - RESIDUES

    KW - URINE

    KW - LEVEL

    M3 - Article

    VL - 70

    SP - 4369

    EP - 4375

    JO - Analytical Chemistry

    JF - Analytical Chemistry

    SN - 0003-2700

    IS - 20

    ER -