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

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，5 μm），以甲醇-0.1%甲酸水为流动相，梯度洗脱，流速0.2 mL·min^-1，柱温30℃，进样量5 μL；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·mL^-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.

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