基于UPLC-TQ-MS考察不同树龄银杏叶双黄酮含量变化规律

Determination of Biflavones in Ginkgo biloba Leaves of Different Ages by UPLC-TQ-MS

目的:建立超高效液相色谱串联三重四级杆质谱法同时测定银杏叶中5个双黄酮成分的含量。方法:采用UPLC BEH C18色谱柱(2. 1 mm×100 mm,1. 7μm),流动相0. 10%甲酸溶液(A)-乙腈(B),梯度洗脱(0~3 min,60%A;3~4 min,60%~50%A;4~6 min,50%A;6~8 min,50%~0A),流速0. 3 m L·min-1,柱温35℃,进样量1μL。结果:穗花杉双黄酮、白果黄素、银杏素、异银杏素、金松双黄酮分别在0. 02~13. 20 mg·L-1(r=0. 996 3),0. 05~23. 60 mg·L-1(r=0. 995 5),0. 09~18. 60 mg·L-1(r=0. 992 7),0. 10~21. 00 mg·L-1(r=0. 998 8)和0. 06~16. 00 mg·L-1(r=0. 996 7)线性关系良好。5个双黄酮加样回收率分别为101. 50%,98. 78%,97. 59%,97. 24%,101. 09%,RSD分别为2. 7%,2. 7%,3. 1%,2. 8%,1. 3%。银杏叶中双黄酮含量差异性较大,穗花杉双黄酮、白果黄素、银杏素、异银杏素、金松双黄酮的质量分数分别为121. 30~434. 74,268. 39~847. 14,251. 80~1 297. 10,195. 87~691. 10,477. 48~3 003. 90μg·g-1,总双黄酮质量分数1 474. 45~5 635. 40μg·g-1,但显示具有一定的规律性,即低树龄银杏叶含总双黄酮较高,随着树龄的增长,总双黄酮含量逐渐降低。结论:该方法专属性强,为银杏叶双黄酮类化合物的质量控制提供了一定的参考。

Objective:To establish an ultra-high performance liquid chromatography coupled with triple quadrupole mass for the determination of five biflavones.Method:Chromatographic separation was carried out on a ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7μm)with gradient elution of acetonitrile and 0.10%formic acid at a flow rate of 0.3 m L·min-1,and the column temperature was set at 35℃.Result:Amentoflavone,bilobetin,ginkgetin,isoginkgetin,sciadopitysin showed a good linearity within the ranges of0.02-13.20 mg·L-1(r=0.996 3),0.05-23.60 mg·L-1(r=0.995 5),0.09-18.60 mg·L-1(r=0.992 7),0.10-21.00 mg·L-1(r=0.998 8),0.06-16.00 mg·L-1(r=0.996 7),with average recoveries of 101.50%,98.78%,97.59%,97.24%,101.09%,and RSDs of 2.7%,2.7%,3.1%,2.8%,1.3%.The contents of amentoflavone,bilobetin,ginkgetin,isoginkgetin,sciadopitysin ranged between 121.30-434.74,268.39-847.14,251.80-1 297.10,195.87-691.10,477.48-3 003.90μg·g-1.The total biflavones ranged between1 474.45-5 635.40μg·g-1.It shows a certain regularity that the low-vinegar leaves contain higher total flavonoids,and the total flavonoid content gradually decreases with the increase of tree age.Conclusion:The method was simple,and can be used for qualitative and quantitative analysis of biflavones.