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首页> 外文期刊>Journal of Lipid Research >A sensitive procedure for the study of beta-carotene-d8 metabolism in humans using high performance liquid chromatography-mass spectrometry.
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A sensitive procedure for the study of beta-carotene-d8 metabolism in humans using high performance liquid chromatography-mass spectrometry.

机译：使用高效液相色谱-质谱法研究人类β-胡萝卜素-d8代谢的灵敏程序。

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This report describes the development of a robust method of high sensitivity for studying the metabolism of beta-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 effluent was nebulized using helium and the solvent was removed under vacuum within the dual-stage particle beam interface. The de-solvated carotenoids were ionized in the negative-ion mode (electron capture) 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 beta-carotene. beta-Carotene-d8 was quantified in the plasma over a concentration range of two orders of magnitude using beta-carotene-(13)C(40) as an internal standard. The overall coefficient of variance (CV) for determining the concentration of the analytes from 30 microliter of plasma was 3.9% for beta-carotene and 2.4% for beta-carotene-d8. Using the LC/PB-MS method, the concentration of beta-carotene-d8 was determined in the plasma of a subject who had consumed a single 5-mg dose over a 30-day period. The sensitive semi-automated procedure is capable of high sample throughput and makes large comprehensive studies feasible.
机译：这份报告描述了一种强大的高灵敏度方法的开发，该方法可使用液相色谱/粒子束质谱（LC / PB-MS）组合研究人类中β-胡萝卜素-d8的代谢。一项初步研究证明了LC / PB-MS方法的实用性。将类胡萝卜素从血浆中提取到己烷中，并使用C-18色谱柱通过反相高效液相色谱（HPLC）进行分离。使用氦气雾化HPLC流出液，并在双级粒子束界面内真空除去溶剂。使用甲烷化学电离以负离子模式（电子捕获）将去溶剂化的类胡萝卜素电离，并使用选定的离子监控器进行检测。该方法的检出限为β-胡萝卜素约0.3 ng（约0.6 pmol）。使用β-胡萝卜素-（13）C（40）作为内标，在两个数量级的浓度范围内对血浆中的β-胡萝卜素-d8进行了定量。用于确定30微升血浆中分析物浓度的总方差系数（CV）对于β-胡萝卜素为3.9％，对于β-胡萝卜素-d8为2.4％。使用LC / PB-MS方法，确定在30天内服用了5毫克单剂的受试者的血浆中β-胡萝卜素-d8的浓度。灵敏的半自动化程序能够实现较高的样品通量，并使大规模的综合研究成为可能。

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