葛根芩连片配伍后葛根素在大鼠体内药动学研究

Pharmacokinetics of puerarin in Gegen Qinlian tablets in compatibility with other drugs

目的建立测定血浆中葛根素浓度的UPLC-MS/MS方法,探究葛根芩连片复方配伍对葛根素在大鼠体内的药动学行为的影响。方法采用甲醇沉淀蛋白法处理血浆样品,以Waters Acquity UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm)进行分离;电喷雾负离子化平行反应离子监测模式检测。结果血浆中葛根素质量浓度在2.500~300.0 ng·mL^(-1)与色谱响应线性关系良好,最低定量限为2.500 ng·mL^(-1),经方法学验证符合生物样本分析要求。大鼠灌胃葛根芩连片、葛根单味药和葛根素对照品后,Cmax分别为(208.2±51.29)、(81.04±11.71)、(88.40±10.79)μg·L^(-1),tmax分别为(0.5±0.0)、(0.6±0.4)、(0.2±0.0)h,t1/2分别为(2.1±0.1)、(1.7±0.1)、(2.0±0.3)h,AUC0~8分别为(186.3±18.98)、(140.0±21.45)、(124.2±44.04)μg·h·L^(-1),AUC0~∞分别为(199.0±16.85)、(144.3±22.42)、(133.1±44.90)μg·h·L^(-1)。结论成功建立了血浆中葛根素的UPLC-MS/MS测定方法,葛根芩连片复方配伍能够改善制剂中主成分葛根素在体内的代谢行为,具有协同增效的作用。

Objective To establish an LC-MS method to determine the concentration of puerarin in the plasma,and determine the effect of compound compatibility of puerarin in Gegen Qinlian tablets on the pharmacokinetics in rats.Methods Plasma samples were separated by Waters Acquity UPLC HSS T3 column(100 mm×2.1 mm,1.8μm)after protein precipitation by methanol.Results Puerarin was determined by negative ion electrospray ionization in parallel reaction monitoring mode.Puerarin in the plasma showed a good linearity at 2.500~300.0 ng·mL^(-1)with an lower limit of quantification of 2.500 ng·mL^(-1).The method validation met the criteria for bio-sample analysis.The main pharmacokinetic parameters of single-dose Gegen Qinlian tablets,Puerariae Lobatae Radix and puerarin in the rats were as follows:Cmax(208.2±51.29),(81.04±11.71),and(88.40±10.79)μg·L^(-1);tmax(0.5±0.0),(0.6±0.4),and(0.2±0.0)h;t1/2(2.1±0.1),(1.7±0.1),and(2.0±0.3)h;AUC0~8(186.3±18.98),(140.0±21.45),and(124.2±44.04)μg·h·L^(-1);AUC0~∞(199.0±16.85),(144.3±22.42),and(133.1±44.90)μg·h·L^(-1).Conclusion The UPLC-MS/MS method for the determination of puerarin in the plasma is established successfully.Compound compatibility of Gegen Qinlian tablets help improve the metabolic behavior of puerarin in vivo,with an obvious synergistic effect.