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一种基于超高效液相色谱-高分辨质谱/质谱的新型生物分析方法,用于定量测定人血清中的橄榄苦苷。应用于药代动力学研究。

A novel bioanalytical method based on UHPLC-HRMS/MS for the quantification of oleuropein in human serum. Application to a pharmacokinetic study.

作者信息

Lemonakis Nikolaos, Mougios Vassilis, Halabalaki Maria, Skaltsounis Alexios-Leandros, Gikas Evagelos

机构信息

Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopoli Zografou, 15771, Athens, Greece.

School of Physical Education and Sports Science, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greece.

出版信息

Biomed Chromatogr. 2016 Dec;30(12):2016-2023. doi: 10.1002/bmc.3779. Epub 2016 Jul 24.

Abstract

A highly sensitive, rapid and specific ultrahigh-performance liquid chromatography, coupled to negative electrospray ionization high-resolution tandem mass spectrometry, method was developed and validated in order to investigate the absorption of dietary oleuropein (OE) in human subjects. Serum samples were collected at predefined time points, after oral administration of an olive leaf extract enriched in OE (204.4 mg OE per capsule) to two subjects. Subsequently, samples were analyzed by the developed method after a simple solid-phase extraction step. Chromatographic separation was operated with aqueous formic acid, 0.1% (v/v), and acetonitrile following a gradient program at a flow rate of 0.45 mL/min in an RP-C (50 × 2.1 mm, 1.9 μm) column with a total run time of 2.7 min. The method was validated and successfully applied to the determination of OE in human serum, with the pharmacokinetic analysis of the data revealing a biphasic response.

摘要

为了研究人体受试者对膳食橄榄苦苷(OE)的吸收情况,开发并验证了一种高灵敏度、快速且特异的超高效液相色谱法,该方法与负电喷雾电离高分辨率串联质谱联用。向两名受试者口服富含OE的橄榄叶提取物(每粒胶囊含204.4毫克OE)后,在预定时间点采集血清样本。随后,经过简单的固相萃取步骤后,用所开发的方法对样本进行分析。在RP-C(50×2.1毫米,1.9微米)柱中,以0.45毫升/分钟的流速,采用含0.1%(v/v)甲酸的水溶液和乙腈按照梯度程序进行色谱分离,总运行时间为2.7分钟。该方法经过验证并成功应用于人体血清中OE的测定,数据的药代动力学分析显示出双相反应。

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