HPLC-DAD-ELSD法同时测定牛黄镇惊丸中5个成分的含量

Simultaneous determination of 5 components in Niuhuang Zhenjing pills by HPLC-DAD-ELSD

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 5μC18(2)色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.2%甲酸溶液为流动相,梯度洗脱,流速1.0 m L·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0.02~0.52μg(r=0.999 9)、0.10~1.90μg(r=0.999 9)、0.03~0.58μg(r=0.999 9)、0.24~4.84μg(r=1.000 0)和0.08~1.62μg(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, 5μm ）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 forprim-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 ofprim-O-glucosylcimifugin,liquiritin, 5-O-methylvisammioside, glycyrrhizic acid and cholic acid were 0.02-0.52 μg （ r=0.999 9 ）, 0.10- 1.90μg（ r=0.999 9 ）, 0.03-0.58μg（ r=0.999 9 ）, 0.24-4.84μg（ r=1.000 0 ）and 0.08-1.62 μg（ r=l.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.