HPLC-DAD-ELSD法同时测定牛黄镇惊丸中5个成分的含量

目的：建立HPLC-DAD-ELSD同时测定牛黄镇惊丸中升麻素苷、甘草苷、5-O-甲基维斯阿米醇苷、甘草酸和胆酸含量的方法，为牛黄镇惊丸的质量评价提供依据。方法：采用Luna 5μ C₁₈（2）色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.2%甲酸溶液为流动相，梯度洗脱，流速1.0 mL·min^-1。升麻素苷、甘草苷、5-O-甲基维斯阿米醇苷和甘草酸的检测波长为254 nm；胆酸使用ELSD检测器，漂移管温度110℃，氮气流速3.2 L·min^-1。结果：升麻素苷、甘草苷、5-O-甲基维斯阿米醇苷、甘草酸和胆酸进样量分别在0.02~0.52 μg（r=0.999 9）、0.10~1.90 μg（r=0.999 9）、0.03~0.58 μg（r=0.999 9）、0.24~4.84 μg（r=1.000 0）和0.08~1.62 μg（r=1.000 0）范围内与峰面积呈现良好的线性关系；平均回收率（n=6）分别为91.4%、101.0%、104.3%、96.4%和98.9%，RSD分别为4.6%、2.4%、3.6%、3.2%和3.1%。测得8批样品中升麻素苷、甘草苷、5-O-甲基维斯阿米醇苷、甘草酸和胆酸的含量分别为0.027~0.113、0.544~0.954、0.046~0.140、0.985~1.672和0.475~1.922 mg·g^-1。结论：本方法可用于牛黄镇惊丸的质量控制。

Objective: To establish an HPLC-DAD-ELSD method for simultaneous determination of prim-O-glucosylcimifugin,liquiritin,5-O-methylvisammioside,glycyrrhizic acid and cholic acid in Niuhuang Zhenjing pills.Methods: HPLC assay was performed on a Luna 5μ C₁₈(2) column(4.6 mm×250 mm,5 μm) with a mixture of acetonitrile and 0.2% formic acid as the mobile phase in gradient elution at a flow rate of 1.0 mL·min^-1.Detection wavelength for prim-O-glucosylcimifugin,liquiritin,5-O-methylvisammioside and glycyrrhizic acid was set at 254 nm and ELSD was used for detection of cholic acid.The temperature of the drift tube was maintained at 110℃ and the nebulizing gas flow rate was 3.2 L·min^-1.Results: The linear ranges of prim-O-glucosylcimifugin,liquiritin,5-O-methylvisammioside,glycyrrhizic acid and cholic acid were 0.02-0.52 μg(r=0.999 9),0.10-1.90 μg(r=0.999 9),0.03-0.58 μg(r=0.999 9),0.24-4.84 μg(r=1.000 0) and 0.08-1.62 μg(r=1.000 0),respectively.The average recoveries(n=6) were 91.4% with RSD of 4.6%,101.0% with RSD of 2.4%,104.3% with RSD of 3.6%,96.4% with RSD of 3.2% and 98.9% with RSD of 3.1%,respectively.The content of the above-mentioned components in 8 samples were 0.027-0.113 mg·g^-1,0.544-0.954 mg·g^-1,0.046-0.140 mg·g^-1, 0.985-1.672 mg·g^-1 and 0.475-1.922 mg·g^-1.Conclusion: The developed method could be used in quality control of Niuhuang Zhenjing pills.