LC-MS/MS法测定不同给药方式下炎琥宁在比格犬体内浓度及药代动力学研究

目的 建立比格犬血浆中炎琥宁的LC-MS/MS测定方法,研究注射给药和口服给药在比格犬体内的药代动力学特征.方法 用单剂量、两制剂、两周期、空腹、交叉试验设计,比格犬6只随机分为2组,分别注射和口服给予20 mg·kg-1炎琥宁.采集不同时间点的血液样本,经沉淀蛋白后,进行LC-MS/MS分析.以乙腈-0.5％甲酸水梯...

Determination of the concentration of potassium sodium dehydroandroandrographolide succinate under different administration methods in Beagle dogs by LC-MS/MS and its pharmacokinetics

Objective To establish a LC-MS/MS method for the determination of potassium sodium dehydroandroandrographolide succinate in plasma of Beagle dogs and to study the pharmacokinetics of potassium sodium dehydroandroandrographolide succinate by injection and oral administration in Beagle dogs.Methods Single dose,two-treatment,two-sequence,two-period,fasting,crossover experiments were designed.Six beagle dogs were randomly divided into two groups.20 mg·kg-1 injection or oral dosage forms of potassium sodium dehydroandroandrographolide succinate were given to each group.The beagle dogs were bled at different time points,after protein precipitation,the samples were analyzed by LC-MS/MS by gradient elution with the molile phase of acetonitrile-0.5%formic acid water.ZORBAX Eclipse Plus C18 column was used for separation.An electrospray ion source was used.Potassium sodium dehydroandroandrographolide succinate was determined under negative ion mode and the internal standard of carbamazepine was monitored by positive ion mode.The blood concentration of potassium sodium dehydroandroandrographolide succinate was determined by multiple reaction monitoring model.The relevant pharmacokinetic parameters were calculated using Win Nonlin 6.0 data statistics software.Results In the range of 10.0-1.0×10^5 ng·mL^-1(r=0.9975),potassium sodium dehydroandroandrographolide succinate had a good linear relationship with the peak area and concentration of internal standard.The lower limit of quantification was 10.0 ng·mL^-1.The pharmacokinetic parameters of potassium sodium dehydroandroandrographolide succinate in Beagle dog plasma were as follows:the t1/2 of potassium sodium dehydroandroandrographolide succinate injection group was(2.56±0.72)h,Cmax was(7.02×10^4±6691.65)ng·mL^-1,AUC0→∞was(1.39×10^4±1568.66)ng·mL^-1·h,Vd was(44.50±14.06)L·mg^-1;the t1/2 of potassium sodium dehydroandroandrographolide succinate oral group was(1.76±0.56)h,Cmax was(351.65±138.53)ng·mL^-1,AUC0→∞was(587.19±266.48)ng·mL^-1·h,Vd was(1252.30±575.81)L·mg^-1,oral absolute bioavailability was(4.16±1.55)%.Conclusion For the first time,this study established a LC-MS/MS analysis method suitable for the determination of the concentration of potassium sodium dehydroandroandrographolide succinate in Beagle dogs,which provides a way for the study of pharmacokinetics in vivo.There are some differences between the kinetic characteristics of potassium sodium dehydroandroandrographolide succinate injection and oral administration,which can provide some reference for the research of the new dosage form of it.