益肾乌发口服液中二苯乙烯苷、大黄素、大黄素甲醚的含量测定

Determination of Stilbene Glucoside,Emodin,Emodin Monomethyl Ether in Yi Shen Wu Fa Oral Liquids by HPLC

目的建立益肾乌发口服液中2,3,5,4′四羟基二苯乙烯-2-O-β-D-葡萄糖苷、大黄素、大黄素甲醚含量的测定方法,从指标成分角度来探讨益肾乌发口服液毒性与何首乌的关系,并为益肾乌发口服液的质量控制提供方法。方法采用HPLC法,C18柱（4.6 mm×250 mm,5μm）;测定二苯乙烯苷的流动相为乙腈-水（25：75）,检测波长为320nm,流速为1.0 mL.min-1,柱温：25℃;测定大黄素大黄素甲醚的流动相为甲醇-0.1%磷酸溶液（80：20）,检测波长为254nm,流速为1.0 mL.min-1,柱温：25℃。结果二苯乙烯苷在0.1μg～10μg、大黄素在0.0168μg～1.68 g、大黄素甲醚在0.0208~1.04μ g范围内呈良好的线性关系,r分别为0.9989（n=8）、0.9984（n=8）、0.9966（n=8）;平均回收率：二苯乙烯苷为99.33%,大黄素为99.998%,大黄素甲醚为99.576%;RSD：二苯乙烯苷为0.68%,大黄素为1.11%,大黄素为1.29%。结论高效液相色谱法测定益肾乌发口服液中二苯乙烯苷、大黄素、大黄素甲醚的含量方法操作简单、灵敏度高、干扰少、重现性好、回收率高,可用于益肾乌发口服液的含量测定。初步证实益肾乌发口服液的毒性可能与君药制何首乌中二苯乙烯苷、大黄素、大黄素甲醚的含量有关,应控制其剂量范围,避免不良反应的发生。

Objective To establish a method for determing 2,3,5,4-tetrahydroxystibene-2-O-β-D-glucosid 、emodin、emodin monomethyl ether in yishen wufa oral liquids.To explore the relationship of toxicity of Yi Shen Wu Fa oral liquids and Radix Polygoni multiflori from the perspective of index constituents,and to provide method for the quality research of yishen wufa oral liquids.Methods HPLC,stationary phase was C18 column（4.6mm×250mm,5μm）,mobile phase of 2,3,5,4′-tetrahydroxystibene-2-O-β-D-glucosid was acetonitrile-water solution（25∶75）,detection wavelength 320nm,flow rate1.0mL·min-1 and column temperature25℃;mobile phase of emodin and emodin monomethly ether was methanol-0.1% phosphric acid（80：20）,detection wavelength 254nm,flow rate1.0mL·min-1 and column temperature25℃.Results There was good linearity in the range of 0.1μg～10μg（r=0.9989） for 2,3,5,4′-tetrahydroxystibene-2-O-β-D-glucosid、in the range of 0.0168～1.68μg（r=0.9984） for emodin and in the range of 0.0208~1.04μg（r=0.9966）emodin monomethly ether;the average recovery of 2,3,5,4′-tetrahydroxystibene-2-O-β-D-glucosid、emodin、emodin monomethyl ether were 99.33%、99.998% and 99.576%,and RSD were 0.68%,1.11 and 1.29%（n=5）.Conclusion This method is simple,with high sensitivity and less interference,high reproducibility and recovery,can be used for quality control of Yi Shen Wu Fa oral liquids.It is initially confirmed that the toxicity of Yi Shen Wu Fa oral liquids may be related with the content of Stilbene glucoside、Emodin and Emodin monomethyl ether in Radix Polygoni multiflori.So,it should control the dose range of Yi Shen Wu Fa oral liquids and avoid adverse events.