UPLC-MS/MS法同时测定比格犬血浆中表儿茶素与4β-(2-氨基乙硫基)表儿茶素

Simultaneous Determination of Epicatechin and 4β-2( aminoethylthio) Epicatechin in Dog Plasma by UPLC-MS/MS

目的:建立UPLC-MS/MS法同时测定比格犬血浆中表儿茶素(EC)与4β-(2-氨基乙硫基)表儿茶素(Cya-EC)的浓度,研究比格犬口服威麦宁胶囊后体内药代动力学。方法:血浆样品经盐酸巯乙胺衍生化处理后,以乙腈"水(0.1%甲酸)为流动相梯度洗脱,Waters BEH Shield RP_(18)(100 mm×2.11 mm,1.7μm)色谱柱分离,采用电喷雾电离源,以多反应监测方式进行正离子检测,定量分析所用的离子反应分别为m/z 291.1→138.9(EC)、m/z366.1→288.9(Cya-EC)和m/z 260.2→116.3(内标:普萘洛尔)。结果:测定血浆中EC及Cya-EC的线性范围均为10~2000 ng·m L^(-1),日内、日间精密度均小于14.82%,准确度均在±4.86%以内;使用DAS3.0药代动力学软件对血药浓度数据进行处理,计算药动学参数,EC及Cya-EC的主要药动学参数t1/2分别为66.3和90.4 h,AUC0-∞分别为2 114.7和12 909.5 ng·h·m L^(-1)。结论:本文建立的UPLC-MS/MS法运行时间短、选择性强、灵敏度高、操作简便、血浆用量少,适用于比格犬口服威麦宁胶囊后体内EC及Cya-EC的药代动力学研究。

Objective： To establish a sensitive and rapid liquid chromatography-tandem mass spectrometry（ LC-MS/ MS） method to simultaneously determinate epicatechin （EC） and 4fl-2 （aminoethylthio） epicatechin （Cya-EC） in plasma of beagle dogs after oral administration of WeiMaiNing capsule. Method： Chromatographic separation was achieved on an Waters BEH Shield RP18 （100 mm ×2. 1 mm, 1.7 μm）using a step gradient program with the mobile phase of 0. 1% formic acid aqueous solution and acetonitrile with 0. 1% formic acid, at a flow-rate of 0. 30 mL·min^-1. Electrospray ionization （ESI）source was applied and operated in the positive ion mode. Multiple reaction monitoring mode with the transitions of m/z 291.1 →138.9 （EC）, m/z 366. 1→ 288.9 （ Cya-EC ） and m/z 260.2 → 116.3 （ IS, propranolol ） was performed to quantify, respectively. Results ： The linear concentration ranges of calibration curves for EC and Cya-EC were 10 - 2000 ng· mL ^- 1. The inter-or intra-day precision（RSD） was less than 14. 82%, and the accuracy （RE）was within ＋ 4. 86%. The pharmacokinetic parameters were calculated by DAS3.0. The t1/2 of EC and Cya-EC were 66. 3 and 90. 4 h. And the AUC0-∞ of EC and Cya-EC were 2 114. 7 and 12 909. 5 ng·h· mL^-1 in beagle dog plasma after oral administration of WeiMaiNing capsules. Conclusion：The method was sensitive, rapid, convenient, using less plasma and was proved to be suitable for preclinical pharmacokinetic studies of EC and Cya-EC in beagle dog after oral administration of Wei MaiNing capsules.