银杏保健品中银杏酸的质谱与色谱法快速分析研究

发布时间：2018-06-08 18:30

  本文选题：实施直接分析-质谱法 + 热辅助在线衍生化-气相色谱法　； 参考：《浙江工业大学》2017年硕士论文

【摘要】：银杏是我国特有的传统植物,富含银杏黄酮和萜内酯等多种生物活性成分,具有很高的食、药用价值,已被开发成各种保健品如银杏茶、白果粉饮品等。本论文的研究对象为银杏的主要毒性成分之一银杏酸,具致敏性、细胞毒性和致突变毒性。欧盟和《中国药典》分别规定银杏叶提取物中银杏酸含量不得超过5mg/kg和10 mg/kg,现行标准中虽未对银杏保健品中的银杏酸含量作出规定,但其摄入量不宜过多。为实现大批量银杏保健品中银杏酸的快速筛查和准确定量,本论文开发了质谱和色谱法用于银杏饮品中银杏酸的分析测定,具体研究成果总结如下:一、建立了实时直接分析-质谱法(Direct Analysis in Real Time-Mass Spectrometry,DART-MS)对白果饮品中的银杏酸进行快速定性以及半定量分析的方法。研究影响离子化效率的因素,确定负离子模式、氦气作为离子化气体、样品移动速度0.2 mm/s、气体温度450℃为最优条件,并进行了方法学考察。该方法在2-50 mg/L浓度范围内线性良好,线性相关系数达0.9981,检出限为0.5mg/L,RSD在12.5%-13.7%(n=6)之间,方法回收率为63.8%-71.2%。以GA13:0为外标,对银杏叶、白果种仁、白果粉饮品中的银杏酸进行半定量分析并辅以二级质谱定性,测得3种样品中总银杏酸含量分别为4799.4±545.3 mg/kg,67.4±10.9 mg/kg和26.6±2.3 mg/kg。将该法和热辅助在线衍生化-气相色谱法的方法学参数、定量结果进行比较,误差在合理范围内,说明DART-MS法适用于银杏酸的快速测定和半定量分析。凭借其快速、实时、前处理步骤少等优点,在大批量银杏保健品中银杏酸的快速筛查、定性以及半定量分析上显示出强大的优势。二、建立了热辅助在线衍生化-气相色谱法(Thermochemolysis-Gas Chromatography,TMC-GC),用于银杏保健品中银杏酸的快速、准确定量。采用甲醇对样品粉末中的银杏酸进行萃取富集,萃取液与三甲基氢氧化硫(Trimethylsulfonium Hydroxide,TMSH)以1:2的比例在裂解炉300℃的高温辅助下瞬间生成银杏酸甲酯,由GC-FID(MS)分离测定,极大地提高了分析灵敏度。结果表明,银杏酸GA13:0在5-600 mg/L浓度范围内线性良好,线性相关系数达0.9994,检出限为1.8 mg/L(S/N=3),RSD为2.4%-2.6%(n=6)。以GA13:0为外标,分别测定白果种仁、白果粉饮品、银杏叶、银杏秋茶和春茶样品中的7种银杏酸,总含量在31.5 mg/kg-1.89×104 mg/kg之间。同时,测定了银杏秋茶和春茶中银杏酸在冲泡90 min后的溶出率分别为16%和12%,溶出量分别为1.95mg/g和2.23 mg/g。白果粉饮品加标回收率在87.1%-106.1%之间。本方法无需耗时的离线衍生化步骤,操作简便、定量准确、灵敏度高,在DART-MS法快速筛查的基础上,本法可进一步应用于大批量样品中银杏酸的准确、快速定量。以上结果表明,本文开发的实时直接分析-质谱法和热辅助在线衍生化-气相色谱法适于银杏保健品中银杏酸的分析研究,结果为银杏保健品的快速筛查和质量控制提供方法参考。
[Abstract]:Ginkgo biloba (Ginkgo biloba) is a unique traditional plant in China. It is rich in many bioactive components such as ginkgo flavonoids and terpene lactones and has high food and medicinal value. It has been developed into various health products such as ginkgo tea and white fruit powder drinks. In this paper, ginkgolic acid, one of the main toxic components of Ginkgo biloba, is studied, which is sensitive, cytotoxic and mutagenic. In EU and Chinese Pharmacopoeia, ginkgolic acid content in ginkgo biloba leaf extract should not exceed 5mg/kg and 10 mg / kg respectively. Although the current standard does not stipulate ginkgolic acid content in ginkgo biloba health products, its intake should not be excessive. In order to screen and quantify ginkgolic acid in large quantities of ginkgo health products, this paper developed mass spectrometry and chromatography for the determination of ginkgolic acid in ginkgo drinks. The specific research results are summarized as follows: 1. A real time direct Analysis in Real time Mass Spectrometry (DART-MS) method was developed for the rapid qualitative and semi-quantitative analysis of ginkgolic acid in ginkgo drinks. The factors affecting the efficiency of ionization were studied. The model of negative ion was determined. The helium as the ionizing gas, the moving speed of the sample was 0.2 mm / s, the temperature of the gas was 450 鈩，

本文编号：1996766