Jin Shu, Zhang Yi-Fan, Chen Xiao-Yan, Liu Ke, Zhong Da-Fang
Soochow University, Suzhou 215123, China.
Yao Xue Xue Bao. 2012 Jan;47(1):105-9.
To study the drug-drug interaction of morinidazole and warfarin and its application, a sensitive and rapid liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of R-warfarin/S-warfarin in human plasma. In a random, two-period crossover study, 12 healthy volunteers received a single oral dose of 5 mg racemic warfarin in the absence and presence of morinidazole. Blood samples were collected according to a pre-designed time schedule. R-warfarin, S-warfarin and methyclothiazide were extracted with ethylether : methylenechloride (3 : 2), then separated on a Astec Chirobiotic V (150 mm x 4.6 mm ID, 5 microm) column using 5 mmol x L(-1) ammonium acetate (pH 4.0) - acetonitrile as mobile phase at a flow-rate of 1.5 mL x min(-1). The mobile phase was splitted and 0.5 mL x min(-1) was introduced into MS. A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM). The resolution of warfarin enantiomers is 1.56. The linear calibration curves for R-warfarin and S-warfarin both were obtained in the concentration range of 5 - 1 000 ng x mL(-1). Intra- and inter-day relative standard deviation (RSD) for R-warfarin and S-warfarin over the entire concentration range across three validation runs was both less than 10%, and relative error (RE) ranged from -4.9% to 0.7%, separately. The method herein described is effective and convenient, and suitable for the study of metabolic interaction between morinidazole and warfarin. The results showed that coadministration of warfarin with morinidazole did not affect the pharmacokinetics of either R-warfarin or S-warfarin.
为研究吗啉硝唑与华法林的药物相互作用及其应用，建立了一种灵敏、快速的液相色谱 - 串联质谱（LC-MS/MS）法测定人血浆中R-华法林/S-华法林。在一项随机、两周期交叉研究中，12名健康志愿者在不存在和存在吗啉硝唑的情况下单次口服5 mg消旋华法林。根据预先设计的时间表采集血样。R-华法林、S-华法林和甲氯噻嗪用乙醚:二氯甲烷（3:2）萃取，然后在Astec Chirobiotic V（150 mm×4.6 mm内径，5 µm）柱上分离，流动相为5 mmol·L⁻¹醋酸铵（pH 4.0）-乙腈，流速为1.5 mL·min⁻¹。流动相分流，0.5 mL·min⁻¹引入质谱仪。配备电喷雾电离源的串联质谱仪用作检测器，在负离子模式下运行。采用多反应监测（MRM）进行定量分析。华法林对映体的分离度为1.56。R-华法林和S-华法林的线性校准曲线均在5 - 1000 ng·mL⁻¹浓度范围内获得。在三个验证批次的整个浓度范围内，R-华法林和S-华法林的日内和日间相对标准偏差（RSD）均小于10%，相对误差（RE）分别为-4.9%至0.7%。本文所述方法有效且方便，适用于吗啉硝唑与华法林代谢相互作用的研究。结果表明，华法林与吗啉硝唑合用不影响R-华法林或S-华法林的药代动力学。
