Automated method for determination of olive oil phenols and metabolites in human plasma and application in intervention studies

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• 作者：M.I. Orozco-Solano ; C. Ferreiro-Vera ; F. Priego-Capote ; M.D. Luque de Castro
• 关键词：Olive oil ; Hydroxytyrosol ; Conjugated phenolic compounds ; Plasma ; LC&ndash ; MS/MS ; On-line SPE
• 刊名：Journal of Chromatography A
• 出版年：2012
• 出版时间：5 October, 2012
• 年：2012
• 卷：1258
• 期：Complete
• 页码：108-116
• 全文大小：649 K

文摘

The interest for olive oil phenols (OOPs) is a growing trend thanks to their contribution to prevent or improve diseases associated to oxidative damage. OOPs ingested in the diet are found at low concentrations in blood either as free forms (e.g. hydroxytyrosol, tyrosol, vanillin, ferulic acid, coumaric acid) or conjugated as sulfate and glucuronide derivatives. Therefore, the identification/quantitation of OOPs in plasma to study their biological effects and elucidate their metabolism requires selective and sensitive methods. The present research describes the development, validation and application of an automated method based on on-line coupling of solid-phase extraction and liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) for quantitation of conjugated and free OOPs in human plasma. This approach minimizes sample handling¡ªthus reducing analyte losses and degradation by contact with the atmosphere¡ªand increases analysis throughput, which is crucial in intervention studies dealing with cohorts formed by numerous individuals. The fundamental of the approach is the retention of OOPs and metabolites in an SPE anionic cartridge with subsequent on-line elution to an LC-MS/MS system. Quantitative analysis of OOPs (relative quantitation for conjugated OOPs) was carried out by selected reaction monitoring mode that reported relative limits of detection and quantitation between 0.02-0.28 ng/mL (16.6-232 pg on-column) and 0.05-0.83 ng/mL (41.5-689 pg on-column), respectively. The accuracy of the method, estimated as recovery factor, ranged from 84.2 to 99.4 % , and precision, expressed as relative standard deviation, was below 3.8 % . The resulting method has been applied to the determination of OOPs and metabolites in plasma samples from individuals who ingested a breakfast prepared with virgin olive oil. The proposed method has an excellent potential for high-throughput use in both clinical and research laboratories.