HPLC法同时测定补肾温肺合剂中6种活性成分含量

Simultaneous Determination of Six Active Compounds in Bushen Wenfei Mixture by HPLC

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.