孕三烯酮胶囊多次给药在巴马小型猪体内的药代动力学研究  

Pharmacokinetics study of gestrinone capsules in Bama minpigs

作　　者：郭湘洁[1] 李钊[1] 谢淑武[1] 周洁芸[1] 李国停 钟瑞华[1] 朱焰[1] 

机构地区：[1]复旦大学生殖与发育研究院上海市计划生育科学研究所生殖药理研究组国家人口和计划生育委员会计划生育药具重点实验室,上海200032

出　　处：《药物分析杂志》2017年第7期1181-1187,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立快速灵敏的HPLC-MS/MS法测定巴马小型猪血浆中孕三烯酮的浓度,并研究孕三烯酮胶囊在巴马小型猪体内的药代动力学特征。方法:以巴马小型猪为实验动物,连续给药12周,每周给药2次,于给药后不同时间点采集血样。通过测定多次给药后小型猪血浆中孕三烯酮的浓度,计算其药代动力学参数。测定血浆样品以孕二烯酮为内标,经环己烷液液萃取后进行LC-MS/MS分析。色谱柱:Agilent ZORBAX Extend-C_(18)(2.1 mm×50 mm,5μm);流动相:水(含0.1%甲酸)-乙腈(含0.1%甲酸),梯度洗脱;流速:0.40 m L·min^(-1);进样量:20μL;柱温:35℃。质谱检测采用ESI正离子模式,扫描方式为多重反应监测(MRM)模式,选择检测离子反应对为m/z 309.2→241.1(孕三烯酮),m/z 311.2→109.1(孕二烯酮)。结果:孕三烯酮在0.1~10 ng·mL^(-1)浓度范围内线性良好(r=0.999 9),定量下限为0.1 ng·mL^(-1)。提取回收率均高于83.1%,日间、日内RSD均小于11.3%,专属性良好,在本实验涉及的条件下样品稳定。8只小型猪首次给药后C_(max)为(2.85±0.96)μg·L^(-1),T_(max)为(2.06±0.62)h,t_(1/2)为(8.43±4.13)h,AUC_(0-85)为(23.81±8.62)μg·L·h^(-1),AUC_(0-∞)为(24.36±8.73)μg·L·h^(-1),MRT_(0-85)为(8.40±2.44)h,MRT_(0-∞)为(9.81±3.88)h;末次给药后C_(max)为(2.70±0.55)μg·L^(-1),T_(max)为(1.69±0.65)h,t_(1/2)为(8.51±3.72)h,AUC_(0-85)为(17.71±4.89)μg·L·h^(-1),AUC_(0-∞)为(18.51±4.48)μg·L·h^(-1),MRT_(0-85)为(8.54±3.50)h,MRT_(0-∞)为(9.98±4.41)h。结论:该方法准确、快速、灵敏,专属性强,适用于动物实验及临床上测定血浆中孕三烯酮的浓度及其药代动力学研究。多次给予孕三烯酮胶囊后2 h血药浓度相差不大,且用药后无不良反应,安全性较高。Objective： To establish a rapid and sensitive HPLC-MS/MS method for the determination of the concentration of gestrinone in Bama minipig plasma, and to study the pharmacokinetic characteristics of gestrinoneCapsules in Bama minipigs. Methods： Bama minipigs were used as experimental animals for 12 weeks and twice a week, the blood samples were collected at different time points after the administration. The pharmacokinetic parameters were calculated by measuring the concentration of gestrinone in plasma of minipigs after muhiple administrations. The plasma samples were extracted by cyclohexane and analyzed by LC-MS / MS. Gestodene was used as the internal standard （ IS ）. Column： Agilent ZORBAX Extend-C18 （ 2.1 mm × 50 mm, 5 μm ） ; mobile phase： water （ containing 0.1% formic acid ）-acetonitrile （ containing 0.1% formic acid ） , gradient elution ; flow rate： 0.40 mL·min^-1, Amount： 20 μL; column temperature ： 35 ℃. Mass spectrometry was performed using ESI positive ion mode, scanning mode was multiplex reaction monitoring （ MRM ） mode, and ion reaction pairs were selected to be m/z 309.2-241.1 （ gestrinone ）, and m/z 311.2-109.1 （ gestodene ）. Results： The concentration of gestrinone in the range of 0.1-10 ng·mL^-1 was linear（ r=0.999 9 ）and the lower limit of quantification was 0.1 ng· mL^-1. The recovery rate was higher than 83.1%, the intra-day and intra-day RSD were less than 11.3%, the specificity was good, and the samples were stable under the conditions involved in this experiment. The first administration pharmacokinetics parameters for gestrinone capsule in 8 minipigs were as followed： Cmax （2.85 ± 0.96 ） μg·L^-1, Tmax （ 2.06 ± 0.62 ） h, t1/2 （ 8.43 ± 4.13 ） h, AUC （ 0-85 ）（ 23.81 ± 8.62 ） μg·L·h^-1, AUC （0-∞）（ 24.36 ± 8.73 ） Iμg·L·h^-1, MRT（0-85） （ 8.40 ± 2.44 ）h, MRT（0-85）（9.81 ± 3.88 ）h; the last administration parameters were as followes ： C （ 2.70 ± 0.55 ） μg·L^-1, Tmax （

关 键 词：孕三烯酮(三烯高诺酮) 液相色谱-质谱联用 巴马小型猪 血药浓度 药代动力学 

分 类 号：R917[医药卫生—药物分析学]