超高效液相-串联质谱法研究银杏叶提取物提取过程中咖啡因的转移规律

张苹苹; 张玉芬; 张晓芳; 冯默; 刘登帅

doi:10.13386/j.issn1002-0306.2019.22.006

超高效液相-串联质谱法研究银杏叶提取物提取过程中咖啡因的转移规律

Transfer Rule of Caffeine in Extraction Processing of Ginkgo biloba leaves Extracts by Ultra-high Performance Liquid Chromatography-tandem Mass Spectrometry

摘要

摘要: 目的:研究银杏叶提取物提取过程中咖啡因的转移规律,采用超高效液相色谱-串联质谱法测定咖啡因含量。方法:色谱柱为ACQUITY UPLC BEH C18(2.1 mm×50 mm,2.7 μm),流动相采用0.1%甲酸溶液和0.1%甲酸乙腈溶液梯度洗脱进行,流速0.3 mL/min,进样量5 μL,柱温30℃,运行时间5 min。质谱参数采用电喷雾离子源(ESI),检测方式采用多反应监测和正离子模式扫描,并进行定量分析。结果:通过优化检测方法,加标回收率在95.23%~102.49%之间,检出限为5 μg/kg,定量限为15 μg/kg。结论:建立的超高效液相色谱-串联质谱检测咖啡因含量方法操作简便,灵敏度高。研究发现,银杏叶中天然存在咖啡因,且不同产地银杏叶中咖啡因含量不同,可能和银杏生长的气候和地域有关系。银杏叶提取生产加工时,原料中约35%的咖啡因会转移至提取物中。

Abstract: Objective:To study transfer rule of caffeine in extraction processing of Ginkgo biloba leaves extracts by an ultra-high performance liquid chromatography-tandem mass spectrometry method. Methods:Chromatographic column was chosen as ACQUITY UPLC BEH C18(2.1 mm×50 mm,2.7 μm). The mobile phase of 0.1% formic acid aqueous solution and 0.1% formic acid acetonitrile was proceeded with gradient elution,at the flow rate of 0.3 mL/min. The chromatographic condition also included sample quantity of 5 μL,column temperature of 30 ℃ and the running time of 5 min. The mass spectra parameters were determined by electrospray ion source(ESI)and multi-reaction monitoring,and the scanning method was positive ion mode. Results:By optimizing the detection method,the recovery was between 95.23% and 102.49%,and the detection limit was 5 μg/kg.The quantitative limit was 15 μg/kg. Conclusion:This UHPLC-MS/MS method was simple and sensitive. It was found that caffeine naturally existed in Ginkgo biloba leaves,and the contents of caffeine in Ginkgo biloba leaves from different origins varied greatly. This might be related to the climate and origin of the growth. During the extraction processing of Ginkgo biloba,thirty-five percent of caffeine in the raw material would be transferred to the extracts.