Development and validation of an LC-MS/MS method for determination of compound K in human plasma and clinical application.
Development and validation of an LC-MS/MS method for determination of compound K in human plasma and clinical application.
复制标题
DOI：
10.5142/jgr.2013.37.135
复制
发表时间：
2013-03
影响因子：
6.3
通讯作者：
Kim MG
中科院分区：
文献类型：
--
作者：
Kim JS;Kim Y;Han SH;Jeon JY;Hwang M;Im YJ;Kim JH;Lee SY;Chae SW;Kim MG
建立了一种快速、灵敏和选择性的分析方法，用于测定人血浆中黄芪皂苷的主要肠道细菌代谢产物化合物K。液-液萃取用于样品制备和分析，然后进行液相色谱串联光谱分析和电喷雾电离接口。采用Phenomenex Luna C18色谱柱（100×2.00 mm，3 μm）分析化合物K，移动的流动相为10 mM乙酸铵-甲醇-乙腈（5：47.5：47.5，v/v/v），流速为0.5 mL/min。对该方法的准确度（相对误差<12.63%）、精密度（变异系数<9.14%）、线性和回收率进行了验证。该测定在校准标准品的整个范围内呈线性，即，浓度范围为1 ng/mL至1，000 ng/ mL（r2 >0.9968）。在1、2、400和800 ng/mL浓度下，液-液萃取后化合物K的回收率分别为106.00± 0.08%、103.50± 0.19%、111.45± 5.21%和89.62±34.46%（日内）和85.40± 0.08%、94.50± 0.09%、112.50± 5.21%、日间分别为95.87±34.46%。人血浆中化合物K分析方法的定量下限为1 ng/ mL。所开发的方法已成功地应用于口服给药后的化合物K在10名健康人受试者的药代动力学研究。
A rapid, sensitive and selective analytical method was developed and validated for the determination of compound K, a major intestinal bacterial metabolite of ginsenosides in human plasma. Liquid-liquid extraction was used for sample preparation and analysis, followed by liquid chromatography tandem spectrometric analysis and an electrospray-ionization interface. Compound K was analyzed on a Phenomenex Luna C18 column (100×2.00 mm, 3 μm) with the mobile phase run isocratically with 10 mM ammonium acetate-methanol-acetonitrile (5:47.5:47.5, v/v/v) at a flow rate of 0.5 mL/min. The method was validated for accuracy (relative error <12.63%), precision (coefficient of variation <9.14%), linearity, and recovery. The assay was linear over the entire range of calibration standards i.e., a concentration range of 1 ng/mL to 1,000 ng/ mL (r2 >0.9968). The recoveries of compound K after liquid-liquid extraction at 1, 2, 400, and 800 ng/mL were 106.00±0.08%, 103.50±0.19%, 111.45±5.21%, and 89.62±34.46% for intra-day and 85.40±0.08%, 94.50±0.09%, 112.50±5.21%, and 95.87±34.46% for inter-day, respectively. The lower limit of quantification of the analytical method of compound K was 1 ng/ mL in human plasma. The developed method was successfully applied to a pharmacokinetic study of compound K after oral administration in ten of healthy human subjects.