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利用超高效液相色谱/电喷雾电离质谱联用技术进行生物分析研究时,绿茶儿茶素快速同步定量方法的开发。

Development of rapid and simultaneous quantitative method for green tea catechins on the bioanalytical study using UPLC/ESI-MS.

作者信息

Misaka Shingen, Kawabe Keisuke, Onoue Satomi, Werba José Pablo, Giroli Monica, Kimura Junko, Watanabe Hiroshi, Yamada Shizuo

机构信息

Department of Pharmacokinetics and Pharmacodynamics and Global Center of Excellence Program, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan.

出版信息

Biomed Chromatogr. 2013 Jan;27(1):1-6. doi: 10.1002/bmc.2740. Epub 2012 Apr 3.

Abstract

A rapid and quantitative analytical method for the simultaneous determination of green tea catechins using ultra-performance liquid chromatography/electrospray ionization-mass spectrometry was developed. Total analytical run time was 3.5 min for the detection of (-)-epicatechin (EC), (-)-epicatechin-3-O-gallate (ECG), (-)-epigallocatechin (EGC), (-)-epigallocatechin-3-O-gallate (EGCG) and myricetin as the internal standard (IS) in rat plasma. The calibration curves were linear over the range of 10-5000 ng/mL for all the catechins. The inter- and intra-day precision (relative standard deviation) and accuracy (percentage deviation) of the method were both lower than 10%. The average extraction recoveries in plasma ranged from 68.5 to 86.5%, and the lower limits of quantification of EC, EGC, ECG and EGCG were 10 ng/mL with a signal-to-noise ratio of >10. The assay developed was successfully applied to a pharmacokinetic study of catechins following intravenous and intragastric administrations of green tea extract in rats. Plasma concentrations of four catechins were detected up to 5-24 h after administration, and the pharmacokinetic parameters of catechins were in agreement with previous studies. From these findings, taken together with the high productivity and precision, the developed method could be a reliable and reproducible tool for the evaluation of pharmacokinetic properties of catechins.

摘要

建立了一种采用超高效液相色谱/电喷雾电离-质谱联用同时测定绿茶儿茶素的快速定量分析方法。在大鼠血浆中检测(-)-表儿茶素(EC)、(-)-表儿茶素-3-O-没食子酸酯(ECG)、(-)-表没食子儿茶素(EGC)、(-)-表没食子儿茶素-3-O-没食子酸酯(EGCG)以及作为内标(IS)的杨梅素时,总分析运行时间为3.5分钟。所有儿茶素的校准曲线在10 - 5000 ng/mL范围内呈线性。该方法的日间和日内精密度(相对标准偏差)以及准确度(偏差百分比)均低于10%。血浆中的平均提取回收率在68.5%至86.5%之间,EC、EGC、ECG和EGCG的定量下限为10 ng/mL,信噪比>10。所建立的测定方法成功应用于大鼠静脉注射和灌胃给予绿茶提取物后儿茶素的药代动力学研究。给药后5 - 24小时检测到四种儿茶素的血浆浓度,儿茶素的药代动力学参数与先前研究一致。基于这些发现,结合高生产率和精密度,所建立的方法可能是评估儿茶素药代动力学特性的可靠且可重复的工具。

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