Determination of tocopherols and their metabolites by liquid-chromatography coupled with tandem mass spectrometry in human plasma and serum.
Talanta, 2017/8/01;170:552-561.
Giusepponi D[1], Torquato P[2], Bartolini D[2], Piroddi M[2], Birringer M[3], Lorkowski S[4], Libetta C[5], Cruciani G[6], Moretti S[1], Saluti G[1], Galli F[2], Galarini R[7]
Affiliations
PMID: 28501210DOI: 10.1016/j.talanta.2017.04.030
Impact factor: 6.556
Abstract
Several studies are increasingly underlying the biological role of vitamin E metabolites as bioactive compounds with anti-inflammatory, anti-proliferative and anti-atherogenic activity. A quantitative method for the simultaneous determination in human plasma and serum of vitamin E (α-tocopherol, α-T and γ-tocopherol, γ-T) and its cytochrome P-450 metabolites: 13'-hydroxychromanol (α-13'-OH), 13'-carboxychromanol (α-13'-COOH) and carboxyethyl hydroxychromanols (α-CEHC and γ-CEHC), was developed and validated. After enzymatic hydrolysis and deproteinization, the metabolites were extracted with a mixture of hexane/ methyl tertiary butyl ether (2/1, v/v). The separation was achieved by reversed phase chromatography and the analytes detected by a triple quadrupole mass analyser using electrospray ionization in positive mode (LC-MS/MS). α-T and γ-T were extracted separately without enzymatic hydrolysis. The analytes were quantified with the isotopic dilution method. After an extensive validation study (three levels in three different occasions for a total of 54 experiments), the procedure was successfully applied to the analysis of sera of healthy volunteers (before and after supplementation with α-T) and plasma of patients affected by chronic kidney disease. Finally, the structures of three unknown compounds found in blood and related to the long chain metabolites (α-13'-OH and α-13'-COOH) were further investigated using liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS).
Keywords: Human plasma/serum; Liquid-chromatography tandem-mass spectrometry (LC-MS/MS); Long chain metabolites; Short chain metabolites; Vitamin E
MeSH terms
Adult; Chromatography, Liquid; Female; Humans; Limit of Detection; Male; Renal Insufficiency, Chronic; Tandem Mass Spectrometry; Tocopherols; Vitamin E; Vitamins
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