液质联用法同时测定加味左金丸中13种有效成分  

Simultaneous determination of 13 bioactive components in Jiawei Zuojin Pill by HPLC-MS/MS

作　　者：陈会朋 王银 陈香玲[2] 丘春晖 余惠旻[3] 张广路[4] CHEN Hui-peng;WANG Yin;CHEN Xiang-ling;QIU Chun-hui;YU Hui-min;ZHANG Guang-lu(Beijing University of Chinese Medicine Shenzhen Hospital(Longgang),Shenzhen 518116,China;Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033,China;Shenzhen University Medical School,Shenzhen 518060,China;Shenzhen Second People's Hospital,Shenzhen 518000,China)

机构地区：[1]北京中医药大学深圳医院(龙岗),广东深圳518116 [2]广东省深圳市中医院,广东深圳518033 [3]深圳大学医学院,广东深圳518060 [4]深圳市第二人民医院,广东深圳518000

出　　处：《中草药》2020年第14期3693-3699,共7页Chinese Traditional and Herbal Drugs

基　　金：深圳市知识创新基础研究(自由探索)(JCYJ20170818094033689);深圳市战略新兴产业发展专项资金(JCYJ20130326112248968);深圳大学聚徒+创研项目(2019延续)。

摘　　要：目的建立了HPLC-ESI-MS/MS法同时测定加味左金丸中阿魏酸、木香烃内酯、黄芩苷、芍药苷、延胡索乙素、吴茱萸次碱、小檗碱、巴马汀、吴茱萸碱、柚皮苷、橙皮苷、柴胡皂苷a与柴胡皂苷d 13种有效成分的分析方法。方法 HPLC采用色谱柱Thermo Syncronis C18柱(100 mm×4.6 mm,3.0μm),流动相为甲醇-0.02 mol/L乙酸铵水溶液,梯度洗脱,体积流量0.35 mL/min,进样量10μL;质谱采用电喷雾离子源、正负离子模式同时采集,通过多反应监测(MRM)同时对加味左金丸中的13种有效成分进行定量分析。结果加味左金丸中13种有效成分阿魏酸、木香烃内酯、黄芩苷、芍药苷、延胡索乙素、吴茱萸次碱、小檗碱、巴马汀、吴茱萸碱、柚皮苷、橙皮苷、柴胡皂苷a与柴胡皂苷d的线性范围分别为2~80μg/mL(r=0.999 2)、0.5~20.0μg/mL(r=0.999 1)、3.5~140.0μg/mL(r=0.999 8)、1~40μg/mL(r=0.999 2)、0.3~12.0μg/mL(r=0.999 1)、1~40μg/mL(r=0.999 2)、3~120μg/mL(r=0.999 7)、2.5~100.0μg/mL(r=0.999 5)、0.5~20.0μg/mL(r=0.999 3)、0.5~20.0μg/mL(r=0.999 1)、1~40μg/mL(r=0.999 1)、0.3~12.0μg/mL(r=0.999 2)、0.3~12.0μg/mL(r=0.999 2),平均加样回收率分别为99.5%、98.9%、100.2%、99.2%、99.5%、99.7%、98.6%、99.9%、101.3%、98.7%、99.8%、97.9%、101.3%,RSD值分别为4.11%、4.88%、1.08%、3.23%、4.13%、3.23%、2.78%、3.12%、4.53%、3.43%、3.58%、5.22%、5.13%。12批加味左金丸样品中13种有效成分的质量分数分别为0.324~0.383、0.051~0.072、3.225~3.466、0.154~0.198、0.015~0.062、0.144~0.199、2.145~2.982、0.441~0.953、0.032~0.099、0.062~0.089、0.111~0.178、0.012~0.065、0.011~0.069 mg/g。结论所建立的分析方法快速、高效、灵敏度高、专属性好,可应用于加味左金丸的质量控制。Objective To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry(HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of 13 major bioactive components(ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a, and saikosaponin d) in Jiawei Zuojin Pills(JZP). Methods The chromatographic separation was performed on a Thermo Syncronis C18 column(100 mm × 4.6 mm, 3.0 μm) with a gradient elution of methanol and 0.02 mol/L ammonium acetate in water at a flow rate of 0.35 mL/min, and the injection volume was 10 μL. The 13 major bioactive components were detected using an electrospray ionization source in ESI+ and ESI- ionization mode, quantified by multiple reaction monitor(MRM) scanning at the same time. Results The linear ranges of ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a and saikosaponin d were 2—80 μg/mL(r = 0.999 2), 0.5—20.0 μg/mL(r = 0.999 1), 3.5—140.0 μg/mL(r = 0.999 8), 1—40 μg/mL(r = 0.999 2), 0.3—12.0 μg/mL(r = 0.999 1), 1—40 μg/mL(r = 0.999 2), 3—120 μg/mL(r = 0.999 7), 2.5—100.0 μg/mL(r = 0.999 5), 0.5—20.0 μg/mL(r = 0.999 3), 0.5—20.0 μg/mL(r = 0.999 1), 1—40 μg/mL(r = 0.999 1), 0.3—12.0 μg/mL(r = 0.999 2), 0.3—12.0 μg/mL(r = 0.999 2), and the average recoveries were 99.5%(RSD = 4.11%), 98.9%(RSD = 4.88%), 100.2%(RSD = 1.08%), 99.2%(RSD = 3.23%), 99.5%(RSD = 4.13%), 99.7%(RSD = 3.23%), 98.6%(RSD = 2.78%), 99.9%(RSD = 3.12%), 101.3%(RSD = 4.53%), 98.7%(RSD = 3.43%), 99.8%(RSD = 3.58%), 97.9%(RSD = 5.22%), and 101.3%(RSD = 5.13%), respectively. The contents of 12 batches of the 13 major bioactive components were 0.324—0.383, 0.051—0.072, 3.225—3.466, 0.154—0.198, 0.015—0.062, 0.144—0.199, 2.145—2.982, 0.441—0.953, 0.032—0.099, 0.062—0.089, 0.111

关 键 词：高效液相色谱-串联质谱 加味左金丸 黄芩苷 芍药苷 阿魏酸 柚皮苷 橙皮苷 木香烃内酯 延胡索乙素 吴茱萸碱 吴茱萸次碱 柴胡皂苷A 柴胡皂苷D 小檗碱 巴马汀 

分 类 号：R286.02[医药卫生—中药学]