丹葛酚酮胶囊中10种成分在Beagle犬体内的药代动力学和排泄研究

Pharmacokinetics and Excretion of Ten Compounds in Dangefentong Capsules in Beagle Dogs

目的:采用超高效液相色谱-串联质谱(UPLC-MS/MS),建立Beagle犬血浆、尿液和粪便中葛根素-6''-O-木糖苷、葛根素芹菜糖苷、3'-甲氧基葛根素、3'-羟基葛根素、葛根素、丹参素、丹酚酸B、迷迭香酸、隐丹参酮、丹参酮ⅡA10种成分的分析方法。研究单次灌胃丹葛酚酮胶囊后上述10种成分在Beagle犬体内的药动学特征和排泄规律。方法:Beagle犬单次灌胃丹葛酚酮胶囊后,分别在0.5 h~24 h内多时间点采集前肢隐静脉血,在48 h内每隔12 h收集尿液和粪便,以UPLC-MS/MS法测定Beagle犬血浆中10种成分的血药浓度,在粪便和尿液中的含量,绘制药-时曲线,计算药动学参数、尿液和粪便排泄率。结果:丹葛酚酮胶囊10种成分的线性相关系数均大于0.99,血浆、尿液和粪便测定精密度与准确度、稳定性、回收率及基质效应等符合生物样品分析要求。5种葛根黄酮T_(max)为2.17 h~2.58 h,t_(1/2)为3.43 h~4.64 h,MRT_(0-t)为5.79 h~6.22 h;4种丹参成分T_(max)为1.08 h~2.88 h,t_(1/2)为3.24 h~7.45 h,MRT_(0-t)为3.44 h~5.98 h。尿液中葛根黄酮总排泄率为0.62%~0.85%,丹参素总排泄率为1.67%,其余丹参成分总排泄率为0.02%~0.16%;粪便中葛根黄酮总排泄率为13.79%~18.52%,丹参素总排泄率为84.05%,其余丹参成分总排泄率为4.36%~15.23%。结论:本方法可用于Beagle犬血浆、尿液、粪便中丹葛酚酮胶囊10种成分测定,葛根黄酮成分在Beagle犬血浆中AUC较大,丹参成分组织分布广泛;10种成分在粪便中排泄量远大于尿液,丹参素排泄率很大,可能是其他丹参酚酸成分的代谢产物。

Objective:To develop a UPLC-MS/MS method for the quantitative determination of puerarin-6′′-O-xyloside,puerarin apioside,3′-methoxy puerarin,3′-hydroxy puerarin,puerarin,tanshinol,salvianolic acid B,rosmarinic acid,cryptotanshinone,and tanshinoneⅡA in the plasma,urine,and feces of Beagle dogs,and investigate the pharmacokinetics characteristics and excretion regularity of the above ten components in Beagle dogs after a single dose of Dangefentong Capsules by gavage.Methods:After a single oral administration of Dangefentong Capsules in Beagle dogs,blood was sampled from the saphenous vein of the forelimb at multiple time points during 0.5 h~24 h,while urine and feces were collected every 12 h within 48 h.The concentrations of ten components in the plasma,feces,and urine were determined by UPLC-MS/MS method.The pharmaceutical concentration-time curve was plotted,and the relevant pharmacokinetic parameters and the excretion rates of urine and feces were calculated.Results:The linear correlation coefficients of 10 components of Dangefentong Capsules were higher than 0.99.The precision,accuracy,stability,recovery,and matrix effect in the determination of plasma,urine,and feces met the requirements of biological sample analysis.The T_(max),t_(1/2),and MRT_(0-t)of the five flavonoids in Puerariae Lobatae Radix were 2.17 h~2.58 h,3.43 h~4.64 h,and 5.79 h~6.22 h,respectively.The T_(max),t_(1/2),and MRT_(0-t)of the four components in Salviae Miltiorrhizae Radix et Rhizoma were 1.08 h~2.88 h,3.24 h~7.45 h,and 3.44 h~5.98 h,respectively.The total excretion rates of flavonoids in Puerariae Lobatae Radix,tanshinol,and other components in Salviae Miltiorrhizae Radix et Rhizoma in the urine were 0.62%~0.85%,1.67%,and 0.02%~0.16%,respectively.The total excretion rates of flavonoids in Puerariae Lobatae Radix,tanshinol,and other components in Salviae Miltiorrhizae Radix et Rhizoma in the feces were 13.79%~18.52%,84.05%,and 4.36%~15.23%,respectively.Conclusion:The method can be used for the determination of ten components of Dangefentong Capsules in the plasma,urine,and feces of Beagle dogs.Flavonoids in Puerariae Lobatae Radix show a high AUC value in the plasma of Beagle dogs,and components in Salviae Miltiorrhizae Radix et Rhizoma are distributed widely.The content of 10 components in feces is significantly higher than that in the urine.The excretion rate of tanshinol is high,which is presumedly a metabolite of other phenolic acids in Salviae Miltiorrhizae Radix et Rhizoma.