Objective: To develop a liquid chromatographic-tandem mass spectrometric method (LC-MS/ MS) for simultaneous determination of silybin and danshensu in rat plasma, and to applied this method to investigate the pharmacokinetics of silybin and Danshensu in rats after a single intravenous administration of silybin-Dan- shensu sodium compound injection as well. Methods: Silybinin, Danshensu and protocatechuic acid (as an internal standard) were extracted from plasma with liquid-liquid extraction by ethyl acetate, and then separated on a Zorbax Eclipse XDB-C18 ( 150 mm ×4.6 mm, 3.5 μm) column using the mobile phase consisted of methanol-0.2％ formic acid (80: 20). The electrospray ionization (ESI) source was applied and operated under the selected reaction monitoring (SRM) mode. Silybin was detected at m/z 480.6→300.9, Danshesnu at m/z 196.8→135.1 and protocatechuic acid at m/z 153.0→109.1. Results: The linear calibration curves were obtained at the concentration ranges of 10 ～ 10 000 ng· mL-1 and 20 ～ 20 000 ng· mL-1 for silybin and D anshensu, respectively. The lower limits of quantitation (LLOQ) were 10 ng· mL -1 and 20 ng· mL-1 for silybin and Danshensu, respectively. The precisions (inter-and intra-day), accuracy (relative error, RE) and stability of the analytes met the requirements. After intravenous administration of silybin-Danshensu sodium compound injection in rats, the pharmacokinetic parameters of silybin were as follows: the t1/2 were ( 2.30 ± 0.45 ), (2.11 ± 0.43 ) and ( 2.40 ± 0.36 ) h; the Ke were (0.31 ±0.06), (0.34 ±0.07) and (0.30 ±0.04) h-1; the AUC0～8h were (3 264.0 ±789.4), (7 706.1 ± 1 750.0) and (21 171.1 ±5 018.7) ng·h·mL -1; the AUC0～∞ were (3 330.3 ±800.1), (7 899.5 ± 1 750.0) and (21 791.0 ± 5 048.1 ) ng· h· mL-1, respectively. The pharmacokinetic parameters of Danshensu were as follows: the t1/2 were (1.98 ±0.47), (1.70 ±0.18) and (2.04 ±0.25) h; the Ke were (0.37 ±0.10), (0.41 ±0.04) and (0.34 ±0.04) h-1; the AUC0～8h were (3758.9 ±1 066.1), (7065.8 ±921.5) and (17228.5 ±3705.2) ng·h·mL-1; the AUC0～∞ were (3 885.8 ±1 152.1), (7 250.7 ±923.1) and (17791.3 ±3720.2) ng·h· mL-1, respectively. Conclusion: This method is highly sensitive and selective, and suitable for the pre-clinical pharmacokinetic study of silybin and Danshensu in compound injection.%目的:建立同时测定大鼠血浆中水飞蓟宾和丹参素浓度的液相色谱-串联质谱法(LC-MS/MS),并用于水飞蓟宾-丹参素钠复方注射剂在大鼠体内的药动学研究.方法:血浆样品经乙酸乙酯液-液萃取后,以甲醇-0.2%甲酸水溶液(80:20)为流动相,Zorbax Eclipse XDB-C18(150 mm×4.6 mm,3.5μm)色谱柱分离,采用电喷雾离子化源(ESI)三重四极杆串联质谱,以选择反应监测模式(SRM)进行检测.用于定量分析的离子反应m/z分别为481.1→300.9(水飞蓟宾)、196.9→135.1(丹参素)和153.0→109.1(内标,原儿茶酸).结果:水飞蓟宾和丹参素分别在10～10 000 ng·mL-1和20～20 000 ng·mL-1浓度范围内呈良好的线性关系,最低定量下限(LLOQ)分别为10和20 ng·mL-1.方法的精密度、准确度和稳定性均符合要求.大鼠静脉注射低、中、高3个剂量的水飞蓟宾-丹参素钠复方注射剂后水飞蓟宾的主要药动学参数为:t1/2分别为(2.30±0.45)、(2.11±0.43)、(2.40±0.36)h,Ke分别为(0.31±0.06)、(0.34±0.07)、(0.30±0.04)h-1,AUC0～8 h分别为(3 264.0±789.4)、(7 706.1±1 750.0)、(21 171.1±5 018.7)ng·h～mL-1,AUC0～∞分别为(3 330.3±800.1)、(7 899.5±1 750.0)、(21 791.0±5 048.1)ng·h·mL-1;丹参素的主要药动学参数为:t1/2分别为(1.98±0.47)、(1.70±0.18)、(2.04±0.25)h,Ke分别为(0.37±0.10)、(0.41±0.04)、(0.34±0.04)h-1,AUC0～8 h分另4为(3 758.9±1 066.1)、(7 065.8±921.5)、(17 228.5±3 705.2)ng·h·mL-1,AUC0～∞分别为(3 885.8±1152.1)、(7 250.7±923.1)、(17 791.3±3 720.2)ng·h·mL-1.结论:本方法具有专属性强、灵敏度高等优点,适用于复方制剂中水飞蓟宾和丹参素的临床前药动学研究.
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