HPLC-ESI-MS/MS同时测定八珍益母丸中9种有效成分
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摘要
目的建立了HPLC-ESI-MS/MS法同时测定八珍益母丸中盐酸水苏碱、盐酸益母草碱、芍药苷、阿魏酸、甘草苷、党参炔苷、毛蕊花糖苷、白术内酯I和猪苓酸C 9种有效成分的分析方法。方法 HPLC法采用色谱柱Waters Atlantis T3柱(150 mm×4.6 mm,3.0μm),流动相为甲醇-0.1%甲酸水溶液,梯度洗脱,体积流量0.5 mL/min,进样量10μL;质谱采用电喷雾离子源、负离子模式(ESI-),通过多反应监测(MRM)同时对八珍益母丸中的9种有效成分进行定量分析。结果八珍益母丸中9种有效成分盐酸水苏碱、盐酸益母草碱、芍药苷、阿魏酸、甘草苷、党参炔苷、毛蕊花糖苷、白术内酯I和猪苓酸C的线性范围分别为0.04～40.00μg/mL(r=0.999 2)、0.04～40.00μg/mL(r=0.999 3)、1.0～100.0μg/mL(r=0.999 1)、0.2～20.0μg/mL(r=0.999 6)、0.2～20.0μg/mL(r=0.997 5)、0.05～5.00μg/mL(r=0.999 4)、0.1～10.0μg/mL(r=0.999 4)、0.1～10.0μg/mL(r=0.999 2)、0.1～10.0μg/mL(r=0.999 2),平均加样回收率分别为99.7%、98.1%、98.5%、101.5%、99.5%、98.4%、99.1%、101.2%、100.1%,RSD值分别为0.52%、0.64%、1.08%、1.12%、0.39%、0.74%、0.91%、0.54%、0.47%。9批八珍益母丸样品中9种有效成分的质量分数分别为0.423～0.752、0.505～0.722、0.613～1.300、0.102～0.184、0.195～0.255、0.021～0.035、0.034～0.072、0.039～0.063、0.051～0.095 mg/g。结论所建立的分析方法简单、灵敏度高、专属性好,可应用于不同厂家、不同生产批次的八珍益母丸中有效成分的测定及质量控制。
Objective To develop and validate a high performance liquid chromatography coupled with electrospray tandem mass spectrometry(HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components(stachydrine hydrochloride,leonurine hydrochloride,paeoniflorin,ferulic acid,liquiritin,lobetyolin,verbascoside,atracylenolide I,and polyporenic acid C) in Bazhen Yimu Pills(BYP).Methods The chromatographic separation was performed on an Waters Atlantis T3 column(150 mm × 4.6 mm,3.0 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.5 mL/min,and the injection volume was 10 μL.The nine major bioactive components were detected using an electrospray ionization source in negative ionization mode(ESI-) and quantified by multiple reaction monitor(MRM) scanning at the same time.Results The linear ranges of stachydrine hydrochloride,leonurine hydrochloride,paeoniflorin,ferulic acid,liquiritin,lobetyolin,verbascoside,atracylenolide I,and polyporenic acid C were 0.04—40.00 μg/mL(r = 0.999 2),0.04—40.00 μg/mL(r = 0.999 3),1.0—100.0 μg/mL(r = 0.999 1),0.2—20.0 μg/mL(r = 0.999 6),0.2—20.0 μg/mL(r = 0.997 5),0.05—5.00 μg/mL(r = 0.999 4),0.1—10.0 μg/mL(r = 0.999 4),0.1—10.0 μg/mL(r = 0.999 2),0.1—10.0 μg/mL(r = 0.999 2),and the average recoveries were 99.7%(RSD = 0.52%),98.1%(RSD = 0.64%),98.5%(RSD = 1.08%),101.5%(RSD = 1.12%),99.5%(RSD = 0.39%),98.4%(RSD = 0.74%),99.1%(RSD = 0.91%),101.2%(RSD = 0.54%),and 100.1%(RSD = 0.47%),respectively.The content of nine batches of the nine major bioactive components were 0.423—0.752,0.505—0.722,0.613—1.300,0.102—0.184,0.195—0.255,0.021—0.035,0.034—0.072,0.039—0.063,and 0.051—0.095 mg/g,respectively.Conclusion The developed method is simple,specific,and sensitive,and it can be applied for the determination of nine major bioactive components and the quality control of BYP collected from different production batches.
Objective To develop and validate a high performance liquid chromatography coupled with electrospray tandem mass spectrometry(HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components(stachydrine hydrochloride,leonurine hydrochloride,paeoniflorin,ferulic acid,liquiritin,lobetyolin,verbascoside,atracylenolide I,and polyporenic acid C) in Bazhen Yimu Pills(BYP).Methods The chromatographic separation was performed on an Waters Atlantis T3 column(150 mm × 4.6 mm,3.0 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.5 mL/min,and the injection volume was 10 μL.The nine major bioactive components were detected using an electrospray ionization source in negative ionization mode(ESI-) and quantified by multiple reaction monitor(MRM) scanning at the same time.Results The linear ranges of stachydrine hydrochloride,leonurine hydrochloride,paeoniflorin,ferulic acid,liquiritin,lobetyolin,verbascoside,atracylenolide I,and polyporenic acid C were 0.04—40.00 μg/mL(r = 0.999 2),0.04—40.00 μg/mL(r = 0.999 3),1.0—100.0 μg/mL(r = 0.999 1),0.2—20.0 μg/mL(r = 0.999 6),0.2—20.0 μg/mL(r = 0.997 5),0.05—5.00 μg/mL(r = 0.999 4),0.1—10.0 μg/mL(r = 0.999 4),0.1—10.0 μg/mL(r = 0.999 2),0.1—10.0 μg/mL(r = 0.999 2),and the average recoveries were 99.7%(RSD = 0.52%),98.1%(RSD = 0.64%),98.5%(RSD = 1.08%),101.5%(RSD = 1.12%),99.5%(RSD = 0.39%),98.4%(RSD = 0.74%),99.1%(RSD = 0.91%),101.2%(RSD = 0.54%),and 100.1%(RSD = 0.47%),respectively.The content of nine batches of the nine major bioactive components were 0.423—0.752,0.505—0.722,0.613—1.300,0.102—0.184,0.195—0.255,0.021—0.035,0.034—0.072,0.039—0.063,and 0.051—0.095 mg/g,respectively.Conclusion The developed method is simple,specific,and sensitive,and it can be applied for the determination of nine major bioactive components and the quality control of BYP collected from different production batches.
引文
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[20]陈勇川,朱卫民,刘松青.高效毛细管电泳法测定八珍丸和八珍益母丸中芍药苷的含量[J].中国药房,2002,13(7):426-427.
[21]钱鑫,黄婉锋.高效液相色谱法测定八珍益母丸中甘草酸铵的含量[J].中药与临床,2014,5(1):21-22.
[22]张晓霞,李晓妮,胡爽,等.HPLC-ESI-MS/MS同时测定四神丸中9种有效成分[J].中草药,2018,49(9):2070-2075.
[23]马莹,李家春,黄文哲,等.UPLC-MS/MS同时测定桂枝茯苓胶囊中6种三萜酸类成分的含量[J].中国中药杂志,2017,49(17):3368-3373.
[2]佟路芳,毛敏,冯成,等.八珍益母丸研究进展[J].首都食品与医药,2009,16(2):25-27.
[3]曾凡军,韩丽梅,段大航,等.八珍益母丸防治药流后出血时间延长的疗效观察[J].中国民康医学,2006,18(4):312-313.
[4]邹凤鸣.八珍益母丸在口服流产药物中的应用[J].航空航天医学杂志,2009,19(8):144-145.
[5]张引儒.八珍益母丸加减治疗气血虚弱型闭经42例[J].内蒙古中医药,2014,33(24):10.
[6]邱二娟.八珍益母丸联合西药治疗气血两虚型月经不调疗效观察[J].新中医,2014,46(8):105-106.
[7]Li X,Wang B,Li Y,et al.The Th1/Th2/Th17/Treg paradigm induced by stachydrine hydrochloride reduces uterine bleeding in RU486-induced abortion mice[J].JEthnopharmacol,2013,145(1):241-253.
[8]Li X,Yuan F L,Zhao Y Q,et al.Effects of leonurine hydrochloride on medically induced incomplete abortion in early pregnancy rats[J].Eur J Obstet Gynecol Reprod Biol,2011,159(2):375-380.
[9]黄瑾,胡晋红,朱全刚.阿魏酸及其衍生物的药理作用[J].中药材,2001,24(7):522-526.
[10]黄圆圆,张元,康利平,等.党参属植物化学成分及药理活性研究进展[J].贵阳中医学院学报,2017,39(4):93-96.
[11]冯亚龙,赵英永,丁凡,等.茯苓皮的化学成分及药理研究进展(I)[J].中国中药杂志,2013,38(7):1098-1102.
[12]宋光西,马玲云,魏锋,等.苯乙醇苷的分布及药理活性研究进展[J].亚太传统医药,2011,7(4):169-171.
[13]王苗苗.白术内酯I拮抗紫杉醇诱导卵巢癌细胞分泌IL-6和化疗耐受[D].成都:成都中医药大学,2010.
[14]苏国林,刘刚,刘育辰,等.甘草苷的提取纯化方法和药理作用研究进展[J].中国现代中药,2011,13(10):48-51.
[15]蒋歆,丁永辉,魏学冰,等.HPLC-ELSD测定八珍益母丸中水苏碱的含量[J].中成药,2011,33(3):542-544.
[16]王怡君,侯峰.HPLC测定八珍益母丸中芍药苷的含量[J].中成药,2004,26(6):1-2.
[17]马向东.HPLC法测定八珍益母丸中芍药苷和阿魏酸的含量[J].中国药物评价,2014,31(4):193-195.
[18]马晓静,崔业波,高楠楠,等.八珍益母丸中芍药苷的HPLC法测定结果的不确定度评价[J].中国现代中药,2017,19(12):1754-1757.
[19]谭永红,肖为蓉,张梅.八珍益母丸中益母草生物碱的含量测定[J].西南国防医药,1999,9(6):369-371.
[20]陈勇川,朱卫民,刘松青.高效毛细管电泳法测定八珍丸和八珍益母丸中芍药苷的含量[J].中国药房,2002,13(7):426-427.
[21]钱鑫,黄婉锋.高效液相色谱法测定八珍益母丸中甘草酸铵的含量[J].中药与临床,2014,5(1):21-22.
[22]张晓霞,李晓妮,胡爽,等.HPLC-ESI-MS/MS同时测定四神丸中9种有效成分[J].中草药,2018,49(9):2070-2075.
[23]马莹,李家春,黄文哲,等.UPLC-MS/MS同时测定桂枝茯苓胶囊中6种三萜酸类成分的含量[J].中国中药杂志,2017,49(17):3368-3373.