摘要
目的建立专属、准确的LC-MS/MS法同时测定大鼠血浆中小檗碱、巴马汀、延胡索乙素、黄芩苷、黄连碱的质量浓度,并应用于加味左金丸在大鼠体内的药动学研究。方法以白屈菜红碱为内标,采用乙腈(含体积分数0.02%甲酸)沉淀蛋白的方式进行血浆样品预处理,色谱柱为Thermo Syncronis C_(18)(100 mm×2.1 mm, 5μm)柱,保护柱为Security Guard C_(18)(4 mm×3.0 mm, 3μm)柱,流动相为2 mmol·L^(-1)甲酸铵水溶液(含体积分数0.02%甲酸)-乙腈(体积比70∶30),流速为0.2 mL·min^(-1),进样量为5μL。离子源选择电喷雾电离源,采用正离子化方式,扫描方式为选择反应离子监测。结果血浆中小檗碱、巴马汀、延胡索乙素、黄芩苷、黄连碱的线性范围分别为0.50-100μg·L^(-1),0.50-100μg·L^(-1),0.10-20μg·L^(-1),5.0-1 000μg·L^(-1)和0.25-50μg·L^(-1)(r≥0.990 7),日内、日间精密度(RSD)均小于15%,准确度RE在-7.5%-5.3%之间。大鼠灌胃给予加味左金丸药液后,小檗碱、巴马汀、延胡索乙素和黄连碱的吸收速度均较快,tmax约为0.3-2.4 h,但它们的达峰浓度都较低,ρmax约为1.9-14.5μg·L^(-1);黄芩苷的t_(max)和ρ_(max)分别为(6.3±1.5) h和(343.0±102.0)μg·L^(-1);延胡索乙素从体内消除的速度最快,t_(1/2)为(1.3±0.4) h,而黄连碱的消除速度最慢,t_(1/2)为(9.3±6.5) h。结论该方法适用于加味左金丸在大鼠体内的药物动力学研究。
Objective To develop a rapid, effective and exclusive LC-MS/MS method for simultaneous determination of berberine, palmatine, tetrahydropalmatine, baicalin and coptisine in rat plasma and to investigate the pharmacokinetics of them after intragastric administration of Jiaweizuojin Pill.Methods Plasma samples were pretreated by one-step protein precipitation with acetonitrile(including 0.02% formic acid).A Thermo Syncronis C_(18) column(100 mm×2.1 mm, 5 μm) and the guard column Security Guard C_(18) column(4 mm×3.0 mm, 3 μm, Dima) were used for the separation of plasma samples, and the mobile phase system included 2 mmol·L^(-1) aqueous ammonium formate(including 0.02% formic acid)-acetonitrile(V∶V=70∶30) at the flow rate of 0.2 mL·min^(-1),injection volume: 5 μL.The positive ionization mode of electrospray ionization(ESI) source was used by selected reaction monitoring(SRM) scanning.Results The method was fully validated and proved to be linear in the ranges of berberine for 0.50-100 μg·L^(-1),palmatine for 0.50-1 000 μg·L^(-1),tetrahydropalmatine for 0.10-20 μg·L^(-1),baicalin for 5.0-1 000 μg·L^(-1),and coptisine for 0.25-50 μg·L^(-1).The RSD values of the intra-day and inter-day precisions were all less than 15%,the accuracies(RE) were within-7.5%-5.3%.The extraction recoveries, the specificity and matrix effects all met the requirements for bioanalytical method validation.The absorption of berberine, palmatine, tetrahydropalmatine and coptisine was fast after intragastric administration of Jiaweizuojin Pill and their tmax was about 0.3-2.4 h, but their peak concentrations were lower and ρmax was about 1.9-14.5 μg·L^(-1);tmax and ρmax of baicalin were(6.3±1.5) h and(343.0±102.0) μg·L^(-1),respectively.The elimination rate of tetrahydropalmatine was the fastest, with t1/2 of(1.3±0.4) h, while the elimination rate of berberine was the slowest with t1/2 of(9.3±6.5) h.Conclusion The method can be applied to pharmacokinetic study of berberine, palmatine, tetrahydropalmatine, baicalin and coptisine after intragastric administration of Jiaweizuojin Pill in rats.
作者
赵宏兴
王鑫
刘有平
邸欣
ZHAO Hongxing;WANG Xin;LIU Youping;DI Xin(School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2021年第7期668-674,共7页
Journal of Shenyang Pharmaceutical University
