建立HPLC-MS-MS法测定比格犬血浆中盐酸戊乙奎醚浓度的方法.盐酸戊乙奎醚血浆样品经石油醚-乙醚(7:3)混合溶液萃取后进样.色谱柱为Agilent反相XDB-C18柱(4,6 mm×150 mm, 5 μm),流动相为甲醇-5 mmol/L醋酸铵水溶液(90:10,v/v,pH 5.8).采用电喷雾离子源,正离子模式,MRM离子采集方式,采集离子(m/z)盐酸戊乙奎醚316.4-128.2[M+H]+,内标盐酸苯海拉明256.4-167.2[M+H]+.标准曲线在1 ng/mL-1000 ng/mL范围内呈线性,相关系数为0.9988.定量下限为0.05 ng/mL.方法的精密度、准确度和回收率均符合要求.分别给予静脉注射0.5,1和5 mg/kg剂量PH后,主要的药动学参数如下:t1/2α0.33 h,t1/2β 2.44 h,tmax 0.058 h,AUC和Cmax与剂量呈线性递增.三个剂量组均符合二室模型.此方法测定比格犬血浆中盐酸戊乙奎醚浓度灵敏、准确、快速,适用于常规盐酸戊乙奎醚药动学研究.%To develop a fast and sensitive liquid chromatography-mass spectrometry method for the determination of penehyclidine hydrochloride (PH) in beagle dog plasma. PH and diphenhydramine hydrochloride (internal standard, IS) were extracted with a solvent mixture of petroleum ether-ethyl ether (7:3). Chromatographic separation was achieved on a reversed-phase Eclipse XDB-C18 column (4.6 mm×l50 ram, 5 μm) using the eluent of methanol-water (5 mmol/L ammonium acetate) (90:10, v/v, pH 5.8) as mobile phase. The electrospray ionization source was set at the positive multiple reaction monitoring (MRM) mode. This method involved the use of the [M+H]+ions of PH and diphenhydramine hydrochloride at m/z 316.4-128.2 and m/z 256.4-167.2. The calibration curve was linear in the range of 1-1000 ng/mL with a correlation coefficient of 0.9988. The lower limit of quantification was 0.05 ng/mL. The precision, accu- racy and recovery of the method were acceptable. Following intravenous injection administration at doses of 0.5, 1 and 5 mg/kg PH, the main pharmacokinetic paraneters were as the followings, t1/2α 0.33 h, t1/2β 2.44 h,tmax 0.058 h, AUC and Cmax exhibited a linear increase along with the increase of dose. The two-compartment model fit the three dose groups. This method was sensitive, accurate and fast for the determination of concentration of PH in beagle dog plasma. It could be used in pharmacokinetic studies of PH.
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