Heinig Uwe, Aharoni Asaph
Department of Plant Sciences, Weizmann Institute of Science, 243 Hertzl Street, Rehovot, 76100, Israel.
Methods Mol Biol. 2014;1153:171-85. doi: 10.1007/978-1-4939-0606-2_12.
Plants of the Solanaceae family are renowned for the production of cholesterol-derived steroidal glycosides, including the nitrogen containing glycoalkaloids and steroidal saponins. In this chapter we describe the use of UPLC (Ultra Performance Liquid Chromatography) coupled with qTOF (Quadrupole Time-of-Flight) mass spectrometry for profiling of these two large classes of semipolar metabolites. The presented method includes an optimized sample preparation protocol, a procedure for high resolution chromatographic separation and metabolite detection using the TOF mass spectrometer which provides high resolution and mass accuracy. A detailed description for non-targeted data analysis and a strategy for putative identification of steroidal glycosides from complex extracts based on interpretation of mass fragmentation patterns is also provided. The described methodology allows profiling and putative identification of multiple steroidal glycoside compounds from the assortment of Solanaceae species producing these molecules.
茄科植物以产生胆固醇衍生的甾体糖苷而闻名，包括含氮的糖生物碱和甾体皂苷。在本章中，我们描述了使用超高效液相色谱（UPLC）与四极杆飞行时间（qTOF）质谱联用，对这两大类半极性代谢物进行分析。所介绍的方法包括优化的样品制备方案、高分辨率色谱分离程序以及使用提供高分辨率和质量准确度的飞行时间质谱仪进行代谢物检测。还提供了非靶向数据分析的详细描述以及基于质量碎片模式解释从复杂提取物中推定鉴定甾体糖苷的策略。所描述的方法能够对产生这些分子的茄科植物种类中的多种甾体糖苷化合物进行分析和推定鉴定。
