HPLC法同时测定补肾温肺合剂中6种活性成分含量
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摘要
目的:建立高效液相色谱法同时测定补肾温肺合剂中莫诺苷、马钱苷、升麻素苷、5-O-甲基维斯阿米醇苷、淫羊藿苷和五味子醇甲6种活性成分的含量。方法:采用Phenomenex luna C18色谱柱(250mm×4.6mm,5.0μm),以甲醇(A)-0.1%甲酸水溶液(B)为流动相,线性梯度洗脱(0～20min,45%A;20～21min,45%A→65%A;21～30min,65%A;30～35min,65%A→67%A),流速为1.0mL·min~(-1),检测波长为240nm(莫诺苷和马钱苷)、250nm(五味子醇甲)、254nm(升麻素苷和5-O-甲基维斯阿米醇苷)和270nm(淫羊藿苷),柱温为30℃。结果:莫诺苷、马钱苷、升麻素苷、5-O-甲基维斯阿米醇苷、淫羊藿苷和五味子醇甲分别在0.013 2～0.264、0.010 4～0.208、0.015 6～0.312、0.011 6～0.232、0.01～0.2和0.01～0.2mg·mL~(-1)浓度范围内与其峰面积积分呈良好的线性关系,平均回收率(n=9)分别为100.9%、100.5%、101.0%、99.2%、98.2%和102.5%。3批补肾温肺合剂中莫诺苷、马钱苷、升麻素苷、5-O-甲基维斯阿米醇苷、淫羊藿苷和五味子醇甲的含量范围分别为2.26～2.33、1.12～1.15、2.75～2.91、0.43～0.48、0.31～0.34和0.16～0.19mg·mL~(-1)。结论:经方法学验证,该研究建立的方法可作为补肾温肺合剂质量控制的有效方法。
Objective:To establish an HPLC method for simultaneous determination of morroniside,loganin,Prim-o-glucosylcimifugin,5-O-methylvisammioside,icariin and schisandrin in Bushen Wenfei mixture.Methods:The chromatographic separation was performed on a Phenomenex Luna C18 column(250 mm×4.6 mm,5μm)with the mobile phase of methanol(A)-0.1%aqueous formic acid(B)by gradient elution(0～20 min,45%A;20～21 min,45%A→65%A;21～30 min,65%A;30～35 min,65%A→67%A).The flow rate was 1.0 mL·min~(-1).The detection wavelengths were 240 nm for morroniside and loganin,250 nm for schisandrin,254 nm for Primo-glucosylcimifugin and 5-O-methylvisammioside,270 nm for icariin.The column temperature was controlled at 30℃.Results:Morroniside,loganin,Prim-o-glucosylcimifugin,5-O-methylvisammioside,icariin and schisandrin exhibited good linearity among the ranges of 0.0132～0.264,0.0104～0.208,0.0156～0.312,0.0116～0.232,0.01～0.2 and 0.01～0.2 mg·mL~(-1) respectively;the average recoveries(n=9)were 100.9%,100.5%,101.0%,99.2%,98.2% and 102.5%.The content ranges of morroniside,loganin,Prim-o-glucosylcimifugin,5-O-methylvisammioside,icariin and schisandrin were 2.26～2.33,1.12～1.15,2.75～2.91,0.43～0.48,0.31～0.34和0.16～0.19 mg·mL~(-1).Conclusion:Conclusion:The HPLC method is proved by methodology validation that it can be used as quality control method of Bushen Wenfei mixture.
Objective:To establish an HPLC method for simultaneous determination of morroniside,loganin,Prim-o-glucosylcimifugin,5-O-methylvisammioside,icariin and schisandrin in Bushen Wenfei mixture.Methods:The chromatographic separation was performed on a Phenomenex Luna C18 column(250 mm×4.6 mm,5μm)with the mobile phase of methanol(A)-0.1%aqueous formic acid(B)by gradient elution(0～20 min,45%A;20～21 min,45%A→65%A;21～30 min,65%A;30～35 min,65%A→67%A).The flow rate was 1.0 mL·min~(-1).The detection wavelengths were 240 nm for morroniside and loganin,250 nm for schisandrin,254 nm for Primo-glucosylcimifugin and 5-O-methylvisammioside,270 nm for icariin.The column temperature was controlled at 30℃.Results:Morroniside,loganin,Prim-o-glucosylcimifugin,5-O-methylvisammioside,icariin and schisandrin exhibited good linearity among the ranges of 0.0132～0.264,0.0104～0.208,0.0156～0.312,0.0116～0.232,0.01～0.2 and 0.01～0.2 mg·mL~(-1) respectively;the average recoveries(n=9)were 100.9%,100.5%,101.0%,99.2%,98.2% and 102.5%.The content ranges of morroniside,loganin,Prim-o-glucosylcimifugin,5-O-methylvisammioside,icariin and schisandrin were 2.26～2.33,1.12～1.15,2.75～2.91,0.43～0.48,0.31～0.34和0.16～0.19 mg·mL~(-1).Conclusion:Conclusion:The HPLC method is proved by methodology validation that it can be used as quality control method of Bushen Wenfei mixture.
引文
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[10]舒可成,刘洁.HPLC测定枣仁安神液中的五味子醇甲[J].药物分析杂志,2012,32(8):1414-1416.
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[2]国家药典委员会.中国药典[M].北京:人民卫生出版社,2015:27,66,149,327.
[3]许方敏,薛瑞,叶洪涛,等.五味子醇甲对小鼠抑郁样行为的影响[J].中国药理学与毒理学杂志,2017,31(3):244-249.
[4]赵文望,皮文霞,蔡宝昌,等.马钱苷、莫诺苷对高糖致心肌细胞损伤的保护机制研究[J].中成药,2016,38(1):160-163.
[5]汪铜芳,王建平,潘金火,等.升麻素苷和5-O-甲基维斯阿米醇苷大鼠在体鼻循环吸收研究[J].中国中药杂志,2017,42(9):1772-1776.
[6]刘利利,刘颖新,毛群芳,等.HPLC法同时测定苍术-玄参药对中4种活性成分的含量[J].药物分析杂志,2016,36(1):81-85.
[7]刘颖新,宾子由,王丹,等.高效液相色谱法分析株洲市中药代煎质量[J].中国医院药学杂志,2017,37(23):2359-2363.
[8]李润泽,常增荣,傅欣彤,等.HPLC法测定杞菊地黄丸中莫诺苷、马钱苷和丹皮酚的含量[J].药物分析杂志,2015,35(2):351-354.
[9]姜明明,杨更亮,刘海燕,等.RP-HPLC法测定畅鼻通颗粒中升麻素苷和5-O-甲基维斯阿米醇苷的含量[J].药物分析杂志,2008,28(9):1459-1461.
[10]舒可成,刘洁.HPLC测定枣仁安神液中的五味子醇甲[J].药物分析杂志,2012,32(8):1414-1416.
[11]廖彩霞,戴德雄,诸葛周.HPLC法测定参芪五味子分散片中五味子醇甲[J].中草药,2012,43(3):505-506.
[12]郑林,刘毅,卢锦辉.HPLC法测定前列舒乐颗粒中淫羊藿苷[J].中草药,2010,41(7):1115-1116.
[13]谢香菊,杨清林,王贤英,等.HPLC测定参仙片中淫羊藿苷的含量[J].中国中药杂志,2009,34(15):1984-1985.