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ARS Home Northeast Area Beltsville, Maryland (BHNRC) Beltsville Human Nutrition Research Center Food Composition and Methods Development Laboratory Research Publications at this Location





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Title:   Author  item
                                            Pawlosky, Robertitem
                                            Flanagan, Vincentitem
                                            Novotny, Janet
  Submitted to: Journal of Lipid Research
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 2/17/2000
                                    Publication  Date: 5/1/2000
                                    Citation: Pawlosky, R.J., Flanagan, V.P., Novotny Dura, J. 2000. A sensitive procedure for the study of beta-carotene-d8 metabolism in humans using liquid chromatography-mass spectrometry. Journal of Lipid Research. 41:1027-1031.Interpretive Summary:   This report describes the development of a robust method for   studying the metabolism of (stable hydrogen) deuterium-   labeled-Beta-carotene in humans using liquid chromatography   and mass spectrometry. Deuterium-labeled-Beta-carotene was   extracted from human blood and injected onto a liquid    chromatograph.  The limit of detection was about 0.3 ng for   deuterium-labeled-Beta-carotene.  The compound was   quantified over a concentration range of two orders of   magnitude using an internal standard.  The overall   coefficient of variance (CV) for determining the   concentration of deuterium-labeled-Beta-carotene from blood   was 2.4%.  Using this method, the concentration of   deuterium-labeled-Beta-carotene was determined in the plasma    of a subject who had consumed a single 5 mg dose over a   thirty day period.  The simplified experimental design has   both high sensitivity and a high sample through-put.   Researchers who study human nutrition especially those   interested in how much vitamin-A can be made from Beta-   carotene in humans will use this information to help them   plan studies in different human populations.
                                Technical Abstract:   This report describes the development of a robust method for   studying the metabolism of B-carotene-d8 in humans using a   combination of liquid chromatography/ particle-beam-mass   spectrometry (LC/PB-MS).  The utility of the LC/PB-MS method   was demonstrated in a pilot study.  The carotenoids were    extracted from plasma into hexane and separated by reverse   phase high-performance liquid chromatography (HPLC) using a   C-18 column.  The HPLC solvent was removed under vacuum   within the dual-stage particle-beam interface.  The de-   solvated carotenoids were ionized in the negative-ion mode   using methane chemical ionization and detected using   selected ion monitoring.  The limit of detection of the   method was on the order of 0.3 ng (approximately 0.6 pmol)    for B-carotene.  B-Carotene-d8 was quantified in the plasma   over a concentration range of two orders of magnitude using   B-carotene-13C40 as an internal standard.  The overall   coefficient of variance (CV) for determining the   concentration of the analytes from 30 ul of plasma was 3.9%   for b-carotene and 2.4% for B-carotene-d8.  The simplified   experimental design having both high sensitivity and a high   sample through-put makes large comprehensive studies   feasible.


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