HPLC-ELSD法测定银杏叶提取物及其制剂中银杏内酯A、B、C和白果内酯含量

Determination of Ginkgolide A,B,C and bilobalide in extract of ginkgo biloba leaves and its phytopharmaceutical agents by HPLC-ELSD

目的建立HPLC-ELSD法测定银杏叶提取物（extract of Ginkgo biloba,EGb）及其制剂中银杏内酯A、B、C及白果内酯含量。方法以Kromasil C18为固定相,甲醇-四氢呋喃-水（1∶24∶75）为流动相,流速0.8 ml/min,柱温35℃,蒸发光散射检测器蒸发温度为100℃,气体流速为2.5 L/min,测定EGb及其制剂中银杏内酯A、B、C和白果内酯含量。结果银杏内酯A、B、C、白果内酯的定量限分别为0.106、0.104、0.105、0.105 mg/ml,银杏内酯A、B、C、白果内酯分别在0.127-0.506、0.129-0.548、0.131-0.576、0.131-0.561 mg/ml浓度范围内,其峰面积与浓度的线性关系良好（r〉0.999）,方法回收率为92.4%-100.8%,日内精密度及日间精密度的变异系数分别小于2%、3%。结论HPLC-ELSD法简单、准确,有利于银杏叶提取物中内酯类成分的质量控制。

Objective To establish a high performance liquid chromatography（HPLC） with evaporative light-scattering detector（ELSD） to determine the content of ginkgolide A,B,C and bilobalide in the extract of Ginkgo biloba（EGb） leaves and its pharmaceutical agents.Methods Kromasil C18 was designated as the stationary phase and methanol to tetrahydrofuran to water（1∶24∶75） as the mobile phase;with the column temperature（250 mm×4.6 mm,5 μm） of 35℃ at a flow rate of 0.8 ml/min,the evaporation temperature of 100℃ and the flow rate of gas of 2.5 L/min.Results The limits of quantitation for ginkgolide A,B,C and bilobalide were 0.106,0.104,0.105 and 0.105 mg/ml,respectively.This method is linear over concentration ranges of 0.127-0.506,0.129-0.548,0.131-0.576 and 0.131-0.561 mg/ml for ginkoglide A,B,C and bilobalide,respectively（r〉0.999）.The recoveries for ginkgolide A,B,C and bilobalide were 92.4%-100.8%.The coefficient of variation of intraday and interday precision was respectively less than 2% and 3% for all compounds.Conclusion The validated method is accurate and easy to handle,thus providing the basis for the quality control of EGb.