RP-HPLC法测定银杏叶提取物类脂质体中黄酮和萜类内酯含量的优化方法

Quantitative Determination of Flavonol Glycosides and Terpene Lactones from Ginkgo Leaf Extract Noisome

建立反相高效液相色谱法（RP-HPLC）,测定银杏叶提取物类脂质体（EGBN）中银杏3种黄酮化合物（槲皮素、山奈素、异鼠李素）和4种萜类内酯（银杏内酯A、银杏内酯B、银杏内酯C、白果内酯）的含量,及银杏总黄酮醇苷的饱和溶解度。采用透析法提取出EGBN中的银杏黄酮。黄酮应用Agilent HC-C18色谱柱（4.6mm×250.0mm,5μm）,以V（乙腈）∶V（0.4%磷酸）=28∶72为流动相,流速1.0 mL/min,检测波长360 nm,柱温40℃。萜类内酯应用Agilent HC-C18色谱柱（4.6mm×250.0 mm,5μm）,以V（甲醇）∶V（四氢呋喃）∶V（水）=46∶7∶47为流动相,流速0.8 mL/min,NEB温度60℃,EVA温度105℃。结果表明：透析法可以将EGBN中有效成分很好的分离,从而易于测定其中药物含量。在此色谱条件下,槲皮素、山奈素、异鼠李素分别在2.01~100.50,1.01~50.3,0.26~12.8 mg/mL范围内与峰面积呈良好的线性关系;银杏内酯A、银杏内酯B、银杏内酯C、白果内酯采用外标两点测定;平均加样回收率（n=6）均在（100±2）%范围内;精密度试验的RSD（n=6）均〈2.0%。并将该法应用于银杏总黄酮醇苷的饱和溶解度的测定。该法灵敏、准确、重复性好,结合透析法可以准确的测定EGBN中黄酮、萜类内酯的含量,适合于银杏叶提取物类脂质体的质量控制。

The aim of this study was to develop a simple and accurate isocratic RP-HPLC analytical method to qualify and quantify three flavonol glycosides（Quercetin,Kaempferol and Isorhamnetin） and four terpene lactones（Ginkgolide A,Ginkgolide B,Ginkgolide C and Bilobalide） of ginkgo leaf extract.The contents of flavonoid glycosides and terpene lactones from EGBN were measured by dialysis method.And the saturation solubility of flavonoid glycosides was also measured with this HPLC method.The condition of HPLC method for flavonol glycosides is： column： Agilent HC-C18（4.6 mm×250.0 mm,5 μm）;mobile phase： ACN-0.4% phosphoric acid（28∶72,v/v）.The drug elution was performed at 40 ℃ at a flow rate of 1.0 mL/min and detected at the wavelength of 360 nm.And the condition of HPLC method for terpene lactones is： column： Agilent HC-C18（250.0 mm×4.6 mm,5 μm）;mobile phase： Methonal-THF-Water（46∶7∶47,v/v/v）.The flow rate was kept at 0.8 mL/min.The temperature of NEB and EVA was performed at 60 ℃ and 105 ℃,respectively.The HPLC methods for flavonol glycosides and terpene lactones were shown to be linear,precise,sensitive,selective and accurate.And the contents of flavonoid glycosides and terpene lactones from EGBN can be measured by dialysis method.