HPLC-MS/MS法同时测定不同桂花品种中7个多酚类活性成分的含量

Simultaneous determination of seven active polyphenols in different Osmanthus fragrans cultivars by HPLC-MS/MS

目的:建立同时测定3个桂花品种丹桂、金桂、银桂中7个多酚类活性成分(绿原酸、咖啡酸、毛蕊花糖苷、柚皮苷、芦丁、阿魏酸、槲皮素)的HPLC-MS/MS分析方法。方法:采用Shim-pack VP-ODS色谱柱(2.0 mm×150 mm,5μm),以甲醇-0.1%甲酸水为流动相,梯度洗脱,流速0.2 m L·min^-1,柱温30℃,进样量5μL;质谱采用ESI离子源,多反应监测(MRM)的负离子扫描方式进行检测。结果:绿原酸、咖啡酸、毛蕊花糖苷、柚皮苷、芦丁、阿魏酸、槲皮素的线性范围分别为50~4 000、100~4 000、50~4 000、50~4 000、50~4 000、200~4 000和50~4 000 ng·m L^-1,平均回收率(n=6)分别为96.5%、93.3%、102.6%、97.8%、94.3%、91.5%和101.4%。精密度、重复性和稳定性良好;并符合方法学考察要求。丹桂中上述7个成分含量分别为(8.73±0.12)、(0.44±0.06)、(99.32±1.95)、(0.41±0.22)、(0.42±0.22)、(0.95±0.23)和(2.04±0.08)mg·g^-1,金桂中上述7个成分含量分别为(6.68±0.08)、(0.64±0.02)、(98.67±1.50)、(0.14±0.05)、(1.56±0.08)、(3.88±0.50)和(2.89±0.00)mg·g^-1,银桂中上述7个成分含量分别为(6.00±0.20)、(0.12±0.02)、(73.46±0.14)、(0.16±0.08)、(1.18±0.11)、(0.46±0.01)和(3.38±0.03)mg·g^-1。结论:所建立的方法可用于桂花中绿原酸、咖啡酸、毛蕊花糖苷、柚皮苷、芦丁、阿魏酸和槲皮素7个多酚类活性成分的同时测定。

Objective:To establish an HPLC-MS/MS method for the simultanneous determinnation of seven active ingredients(chlorogenic acid,caffeic acid,verbascoside,naringin,rutin,ferulic acid,and quercetin)in three kinds of Osmanthus fragrans(Dangui,Jingui and Yingui).Methods:The analysis was performed on a Shim-pack VP-ODS column(2.0 mm×150 mm,5 μm)using a mobile phase consisting of methanol and 0.1% formic acid at a flow rate of 0.2 mL·min^-1 by gradient elution.The column temperature was set at 30 ℃,and the injection volume was 5 μL.The target compounds were analyzed by the negative ion multiple reaction monitoring(MRM)mode.Results:The linear ranges of chlorogenic acid,caffeic acid,verbascoside,naringin,rutin,ferulic acid and quercetin were 50-4 000,100-4 000,50-4 000,50-4 000,50-4 000,200-4 000 and 50-4 000 ng·mL^-1,respectively.The average recoveries(n=6)were 96.5%,93.3%,102.6%,97.8%,94.3%,91.5% and 101.4%,respectively.Precision,repeatability and stability were in line with the methodological requirements.The contents of the above seven components in Dangui were(8.73±0.12),(0.44±0.06),(99.32±1.95),(0.41±0.22),(0.42±0.22),(0.95±0.23)and(2.04±0.08)mg·g^-1,respectively.The contents of the above seven components in Jingui were(6.68±0.08),(0.64±0.02),(98.67±1.50),(0.14±0.05),(1.56±0.08),(3.88±0.50)and(2.89±0.00) mg·g^-1;the contents of the above seven components in Yingui were(6.00±0.20),(0.12±0.02),(73.46± 0.14),(0.16±0.08),(1.18±0.11),(0.46±0.01),and(3.38±0.03)mg·g^-1.Conclusion:The established method can be used to determinate seven polyphenols of Osmanthus fragrans simultaneously.