Doheny-Adams T, Redeker K, Kittipol V, Bancroft I, Hartley S E
Department of Biology, University of York, Wentworth Way, York, YO10 5DD UK.
Plant Methods. 2017 Mar 21;13:17. doi: 10.1186/s13007-017-0164-8. eCollection 2017.
Glucosinolates, anionic sulfur rich secondary metabolites, have been extensively studied because of their occurrence in the agriculturally important brassicaceae and their impact on human and animal health. There is also increasing interest in the biofumigant properties of toxic glucosinolate hydrolysis products as a method to control agricultural pests. Evaluating biofumigation potential requires rapid and accurate quantification of glucosinolates, but current commonly used methods of extraction prior to analysis involve a number of time consuming and hazardous steps; this study aimed to develop an improved method for glucosinolate extraction.
Three methods previously used to extract glucosinolates from brassicaceae tissues, namely extraction in cold methanol, extraction in boiling methanol, and extraction in boiling water were compared across tissue type (root, stem leaf) and four brassicaceae species (, , , and ). Cold methanol extraction was shown to perform as well or better than all other tested methods for extraction of glucosinolates with the exception of glucoraphasatin in shoots. It was also demonstrated that lyophilisation methods, routinely used during extraction to allow tissue disruption, can reduce final glucosinolate concentrations and that extracting from frozen wet tissue samples in cold 80% methanol is more effective.
We present a simplified method for extracting glucosinolates from plant tissues which does not require the use of a freeze drier or boiling methanol, and is therefore less hazardous, and more time and cost effective. The presented method has been shown to have comparable or improved glucosinolate extraction efficiency relative to the commonly used ISO method for major glucosinolates in the Brassicaceae species studied: sinigrin and gluconasturtiin in ; sinalbin, glucotropaeolin, and gluconasturtiin in ; glucoraphenin and glucoraphasatin in ; and glucosatavin, glucoerucin and glucoraphanin in .
硫代葡萄糖苷是一类富含阴离子硫的次生代谢产物，因其存在于具有重要农业意义的十字花科植物中以及对人类和动物健康的影响而受到广泛研究。作为一种控制农业害虫的方法，有毒硫代葡萄糖苷水解产物的生物熏蒸特性也越来越受到关注。评估生物熏蒸潜力需要快速准确地定量硫代葡萄糖苷，但目前分析前常用的提取方法涉及许多耗时且危险的步骤；本研究旨在开发一种改进的硫代葡萄糖苷提取方法。
比较了之前用于从十字花科植物组织中提取硫代葡萄糖苷的三种方法，即冷甲醇提取、热甲醇提取和沸水提取，涉及不同的组织类型（根、茎、叶）和四种十字花科植物（[此处原文缺失具体植物名称]）。结果表明，除了[此处原文缺失具体植物名称]茎中的萝卜硫素葡萄糖苷外，冷甲醇提取在提取硫代葡萄糖苷方面的表现与所有其他测试方法相当或更好。还证明了在提取过程中常规用于组织破碎的冻干方法会降低最终硫代葡萄糖苷的浓度，并且在冷的80%甲醇中从冷冻湿组织样品中提取更有效。
我们提出了一种从植物组织中提取硫代葡萄糖苷的简化方法，该方法不需要使用冷冻干燥机或热甲醇，因此危险性更低，更节省时间和成本。相对于研究的十字花科植物中主要硫代葡萄糖苷常用的ISO方法，所提出的方法已被证明具有相当或更高的硫代葡萄糖苷提取效率：[此处原文缺失具体植物名称]中的黑芥子硫苷和葡萄糖芥苷；[此处原文缺失具体植物名称]中的白芥子硫苷、葡萄糖异硫氰酸酯和葡萄糖芥苷；[此处原文缺失具体植物名称]中的萝卜硫苷和萝卜硫素葡萄糖苷；以及[此处原文缺失具体植物名称]中的葡萄糖沙维因、葡萄糖芥碱和萝卜硫苷。
