کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1203499 965115 2011 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Quantitative profiling of phosphatidylethanol molecular species in human blood by liquid chromatography high resolution mass spectrometry
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Quantitative profiling of phosphatidylethanol molecular species in human blood by liquid chromatography high resolution mass spectrometry
چکیده انگلیسی

Phosphatidylethanol (PEth) is a group of aberrant phospholipids formed in cell membranes in the presence of ethanol by the catalytic action of the enzyme phospholipase D on phosphatidylcholine. Recently published literature has demonstrated the existence of several molecular species of PEth in samples drawn from alcohol-dependent subjects. A novel liquid chromatography high-resolution mass spectrometry (LC–HRMS) method coupled with a lipidomic strategy was developed and validated for the quantitative profiling of PEth molecular species in human blood collected from heavy and social drinkers. Chromatography was performed on a C18 column using acetonitrile, 10 mM ammonium acetate, and 2-propanol as mobile phases with a 22-min gradient. HRMS experiments were performed on an LTQ-Orbitrap XL hybrid mass spectrometer equipped with an electrospray ionization source operated in negative ion mode. The theoretical masses of [M−H]− of PEth species were calculated from the elemental chemical formula by varying the length and unsaturation grade of the fatty acid side chains; identification of PEth species in blood was achieved by searching the accurate masses of the targeted compounds in the acquired full-scan LC–HRMS chromatogram. The chemical structure of tentatively identified PEth species was elucidated through HR multiple mass experiments. The validated LC–HRMS method was selective, as warranted by HRMS at 60,000 resolution and 4 ppm accuracy. Linearity was observed in the 0.001–2.000 μM range, and limit of detection of 0.0005 μM and limit of quantitation of 0.001 μM were obtained for single PEth species. Imprecision and inaccuracy were always lower than 10% and 15%, respectively. The identification capabilities of the method were tested on blood samples collected from heavy drinkers (n = 11), social drinkers (n = 8), and teetotalers (n = 10). The high sensitivity of the method led to the simultaneous identification of 17 different PEth molecular species in blood collected from heavy drinkers, and 2 PEth species (16:0/18:1 and 16:0/18:2) in blood collected from social drinkers.


► A LC–HRMS method for the quantitative profiling of PEth homologues in human blood was developed and validated.
► Up to 17 PEth homologues were identified in blood of heavy drinkers.
► PEth 16:0/18:1 and 16:0/18:2 were the major homologues, accounting together for 74% of total PEth in heavy drinkers.
► These homologues were identified also in social drinkers.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1218, Issue 46, 18 November 2011, Pages 8423–8431
نویسندگان
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