产地、树龄、采收期等因素对银杏叶中黄酮和萜内酯含量的影响

Effect of origin,tree age,and harvesting time on content of flavonoids and terpene lactones in Ginkgo Folium

采用高效液相色谱法测定种植基地银杏叶药材中总黄酮醇苷的含量,样品经甲醇-25%盐酸回流提取,Agilent ZORBAX SB-C色谱柱(4.6 mm×250 mm,5μm)分离,以0.4%磷酸水溶液-甲醇(45∶55)为流动相进行等度洗脱,流速1 mL·min^(-1),柱温30℃,检测波长360 nm,进样量10μL。建立了基于超高效液相色谱-三重四极杆/线性离子阱质谱(UPLC-QTRAP-MS/MS)测定银杏叶萜类内酯含量的方法,以乙腈-0.1%甲酸水为流动相进行梯度洗脱,流速0.2 mL·min^(-1),柱温30℃,样品室温度10℃,进样量10μL。质谱条件采用ESI^(+)多反应监测(MRM)模式扫描。分别采用上述方法测定6个银杏基地99批银杏叶样品总黄酮醇苷、萜类内酯含量,并对其结果进行统计分析,比较不同产地、树龄、采收期、性别、加工方式银杏叶中黄酮、萜内酯含量差异。结果表明99批银杏叶样品总黄酮醇苷质量分数为0.38%~2.08%,4种萜内酯总量为0.03%~0.87%。该实验所建立的方法操作简便、结果可靠,可用于银杏叶的定量分析,研究结果为银杏叶的质量控制提供了依据。

The content of total flavonol glycosides in Ginkgo Folium in the planting bases was determined by high performance liquid chromatography(HPLC).The samples were extracted by reflux with methanol-25%hydrochloric acid.The HPLC conditions were as follows:Agilent ZORBAX SB-Ccolumn(4.6 mm×250 mm,5μm),isocratic elution with mobile phase of 0.4%phosphoric acid solution-methanol(45∶55),flow rate of 1 mL·min^(-1),column temperature of 30℃,detection wavelength of 360 nm,and injection vo-lume of 10μL.A method for the determination of terpene lactones in Ginkgo Folium was established based on ultra-high performance liquid chromatograph-triple-quadrupole/linear ion-trap tandem mass spectrometry(UPLC-QTRAP-MS/MS).The UPLC conditions were as below:gradient elution with acetonitrile-0.1%formic acid,flow rate of 0.2 mL·min^(-1),column temperature of 30℃,sample chamber temperature of 10℃,and injection volume of 10μL.The ESI^(+) and multiple reaction monitoring(MRM)were adopted for the MS.The above methods were used to determine the content of total flavonol glycosides and terpene lactones in 99 batches of Ginkgo Folium from 6 planting bases,and the results were statistically analyzed.The content of flavonoids and terpene lactones in Ginkgo Folium from different origins,from trees of different ages,harvested at different time,from trees of different genders,and processed with different methods was compared.The results showed that the content of total flavonol glucosides in 99 Ginkgo Folium samples ranged from 0.38%to 2.08%,and the total content of the four terpene lactones was in the range of 0.03%-0.87%.The method established in this study is simple and reliable,which can be used for the quantitative analysis of Ginkgo Folium.The research results lay a basis for the quality control of Ginkgo Folium.