基于UPLC-MS/MS法研究银杏叶中咖啡因含量的影响因素  

Influencing factors on caffeine content in Ginkgo Folium based on UPLC-MS/MS

作　　者：高陆迎 陈秀芬 杨超杰 石福娟 麦合丽娅·米吉提 魏雪娇 刘春生[1] 杨瑶珺[1] GAO Lu-ying;CHEN Xiu-fen;YANG Chao-jie;SHI Fu-juan;Maiheliya Mijiti;WEI Xue-jiao;LIU Chun-sheng;YANG Yao-jun(School of Chinese Materia Medica,Bejing University of Chinese Medicine,Bejing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《北京中医药》2022年第9期983-989,共7页Beijing Journal of Traditional Chinese Medicine

摘　　要：目的建立超高效液相色谱三重四极杆质谱(UPLC-MS/MS)法,研究不同产地、采收期、树龄、加工方式对银杏叶中咖啡因含量的影响。方法将77批银杏叶样品经甲醇超声提取,蒸干溶剂,酸水溶解后碱化,氯仿萃取3次,ACQUITY UPLC HSS T3色谱柱分离;以0.1%甲酸水溶液、0.1%甲酸乙腈溶液为流动相进行梯度洗脱,采用电喷雾离子源多离子反应监测模式(MRM),外标法定量,研究产地、采收期、树龄、加工方式对银杏叶中咖啡因含量的影响。结果咖啡因含量为0.06~62.50 ng/mL范围时,与其峰面积具有良好的线性关系(R^(2)=0.9999);加样回收率为84.94%,相对标准偏差(RSD)为7.99%。77批样品中均含有咖啡因,咖啡因含量为0.1585~22.1894μg/kg。陕西宁强、福建长汀产银杏叶的咖啡因含量相对较高,江苏邳州、山东郯城、辽宁丹东次之,云南文山较低。结论大多数银杏叶样品中咖啡因含量均稳定在较低水平(<10μg/kg);其含量与银杏叶树龄、采收期、加工方式无明显相关性,而与银杏叶的产地有关。Objective An UPLC-MS/MS method was established to study the effects of different origin,harvest period,tree age and processing methods on caffeine content in Ginkgo Folium.Methods The sample was extracted by ultrasound with methanol,steamed drying solvent,alkalized after dissolution of acid water and chloroform extraction 3 times,and separated by ACQUITY UPLC HSS T3 chromatographic column,with mobile phase of 0.1%formic acid solution-0.1%formic acid acetonitrile solution for gradient elution.Data were conducted at ESI+multiple reaction monitoring(MRM)mode;quantitative analysis was conducted with external standard method.And the effects of origin,harvesting period,tree age and processing method on caffeine content in Ginkgo Folium was studied by statistics.Results With caffeine in the range of 0.06 ng/mL~62.5 ng/mL,there was a good linear relationship between the mass concentration of caffeine and the peak area,R^(2)=0.9999;the sample recovery rate was 84.94%,with RSD of7.99%.The content range of caffeine in 77 batches of Ginkgo Folium sample was 0.1585~22.1894μg/kg.The caffeine content of Ginkgo Folium produced in Ningqiang,Shaanxi province and Changting,Fujian province,was highest,second in Pizhou,Jiangsu Province,Tancheng,Shandong Province and Dandong,Liaoning Province,and lowest in Wenshan,Yunnan Province.Conclusion The content of caffeine in most Ginkgo Folium is stable at a lower level(below 10μg/kg).The content of caffeine in Ginkgo Folium is not significantly related with the tree age,harvesting period and processing method,but with the origin of Ginkgo Folium.

关 键 词：银杏叶 咖啡因 UPLC-MS/MS法 产地 液质联用 

分 类 号：R284.1[医药卫生—中药学]