UPLC-MS/MS法同时测定大鼠血浆中交泰丸主要成分的含量  

A rapid method for simultaneous quantification of main components of Jiaotai Pill in rat plasma using UPLC-MS/MS

作　　者：徐亚杰[1,2] 刘彤 XU Yajie;LIU Tong(Department of Pharmacy,the First Hospital of China Medical University,Shenyang 110001,China;School of Pharmacy,China Medical University,Shenyang 110122,China;Liaoning Institute for Drug Control,Shenyang 110036,China)

机构地区：[1]中国医科大学附属第一医院药学部,辽宁沈阳110001 [2]中国医科大学药学院,辽宁沈阳110122 [3]辽宁省药品检验检测院,辽宁沈阳110036

出　　处：《沈阳药科大学学报》2022年第6期659-665,702,共8页Journal of Shenyang Pharmaceutical University

摘　　要：目的建立超高效液相色谱串联质谱(UPLC-MS/MS)法同时测定大鼠血浆中交泰丸主要活性成分小檗碱、黄连碱、表小檗碱、巴马汀和桂皮醛的浓度,并用于交泰丸活性成分在大鼠体内的血药浓度定量研究。方法以延胡索乙素为内标,采用乙腈沉淀蛋白的方式进行血浆样品预处理,色谱柱为ThermoAcclaim 120-C_(18)(50 mm×4.6 mm,3μm)柱,流动相为乙腈和5 mmol·L^(-1)醋酸铵(体积分数0.05%甲酸),梯度洗脱,流速为0.4 mL·min^(-1),进样量为5μL。离子源选择电喷雾离子化源,采用正离子化方式,扫描方式为多反应离子监测模式。结果血浆中小檗碱在0.20~300.0μg·L^(-1)、黄连碱在0.40~150.0μg·L^(-1)、表小檗碱在0.20~200.0μg·L^(-1)、巴马汀在0.10~100.0μg·L^(-1)以及桂皮醛在0.10~50μg·L^(-1)浓度范围内线性关系良好,相关系数均大于0.993;日内、日间精密度(RSD)均小于15%,准确度RE在-9.6%~8.2%之间;总运行时间为4.5 min,用建立的方法测定了大鼠灌胃给予交泰丸药液1 h后血浆中的小檗碱、黄连碱、表小檗碱、巴马汀和桂皮醛的浓度。结论该方法精密度高、准确度高、操作便捷,适用于交泰丸体内高通量样品分析。Objective To develop a method for simultaneous determination of the active components of Jiaotai Pills,including berberine,coptisine,epiberberine,palmatine and cinnamaldehyde by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)in rat plasma.Methods The plasma samples were pretreated by protein precipitation,and a ThermoAcclaim 120-C_(18)(50 mm×4.6 mm,3μm)column was used for the separation with gradient elution using acetonitrile and 5 mmol·L^(-1)ammonium acetate(0.05%formic acid)as mobile phase.The flow rate was 0.4 mL·min^(-1)and the injection volume was 5μL.The analytes were quantitated by triple quadrupole mass spectrometer with electrospray ionization in positive multiple reaction monitoring(MRM)mode.Tetrahydropalmatine was used as the internal standard.Results The method was fully validated and proved to be linear in the ranges of 0.20-300.0μg·L^(-1)for berberine,0.40-150.0μg·L^(-1)for coptisine,0.20-200.0μg·L^(-1)for epiberberine,0.10-100.0μg·L^(-1)for palmatine and 0.10-50μg·L^(-1)for cinnamaldehyde.All the correlation coefficients were above 0.993.The extraction recoveries,the specificity and matrix effects all met the requirements for bioanalytical method validation.The intra-day and inter-day precision(RSD)were all less than 15%,and the accuracy RE was between-9.6%and 8.2%.The total run-time was 4.5 min.The established method was used to determine the concentrations of berberine,coptisine,epiberberine,palmatine and cinnamaldehyde in plasma of rats after intragastric administration of Jiaotai Pills.Conclusion This method is precise,accurate,convenient and suitable for highthroughput sample analysis.

关 键 词：交泰丸 UPLC-MS/MS 多反应监测 含量测定 血浆 

分 类 号：R917[医药卫生—药物分析学]