甲醇-水流动相HPLC测定银杏萜内酯方法优化

Optimization for Determination Method of Terpene Lactones in Ginko Biloba with Methanol-water as Mobile Phase by HPLC

优化了甲醇-水流动相HPLC测定银杏萜内酯的方法,建立了线性回归方程,并与药典正丙醇-四氢呋喃-水流动相方法进行比较。在此基础上,用两种方法对银杏叶、银杏外种皮、白果以及银杏叶提取物(EGB)样品中的萜内酯含量进行了对照分析。结果表明,甲醇-水梯度洗脱基线噪音小,柱效高,分离时间短。优化的洗脱条件为:甲醇-水为流动相,0~2 min,0~20%甲醇;2~32 min,20%~60%甲醇,建立的方法在50~2000 mg/L的浓度范围内具有良好的线性关系,检出限为0.05~0.09μg,不同浓度水平的加标回收率为92.4%~101.6%,相对标准偏差(RSD)值为2.7%~5.1%。两种方法测定银杏萜内酯没有明显差别,其中测定银杏叶中白果内酯、银杏内酯C、银杏内酯A、银杏内酯B的相对误差分别为0.86、4.2、1.7、3.9。

Optimized HPLC method for determination of terpene lactones in Ginkgo biloba by methanol-water mobile phase.A linear regression equation was established and compared with the pharmacopoeial n-propanol-tetrahydrofuran-water mobile phase method.The content of terpene lactones in Ginkgo leaf,exotestal,seed and extract were compared by these two methods.The results showed the methanol-water gradient elution has lower baseline noise,higher column efficiency and shorter separation time compared with the pharmacopoeia method.The optimized elution conditions:methanol-water as mobile phase,0~2 min,0%~20%methanol;2~32 min,20%~60%methanol.The established method has a good linear relationship in the concentration range of 50~2000 mg/L.The detection limit is 0.05~0.09μg,and the recoveries at different concentration levels is 92.4%~101.6%with RSD 2.7%~5.1%.There was no significant difference in the determination of terpene lactones between these two methods.The relative errors of bibobalide,ginkgolide C,ginkgolide A,and ginkgolide B in Ginkgo leaf were 0.86,4.2,1.7,3.9.