Cornell H J, McLachlan A, Cullis P G
Department of Applied Chemistry, RMIT University, Melbourne, Vic. 3001, Australia.
J Biochem Mol Biol Biophys. 2002 Apr;6(2):151-8. doi: 10.1080/10258140290027315.
Simple extraction of prolamins from cereal flour using 70% aqueous ethanol leads to co-extraction of lipids and other secondary products. Treatment of the crude extract with an excess of pure ethanol resulted in the removal of the majority of these compounds. Prolamin extracts obtained following ethanol precipitation showed little difference to products of more complex, multi stage, selective extractions, when compared using MALDI-TOF mass spectrometry. Toxicity tests on enzymic digests of ethanol-precipitated prolamins from coeliac-toxic cereals, coeliac-non-toxic cereals and oats were carried out using a rat liver lysosome assay. The prolamins from the ethanol-precipitated extracts showed greater activity than those extracted using only 70% ethanol for extraction. As the ethanol precipitation method is simple and provides a prolamin extract of sufficient purity for further evaluation, this procedure has been adopted as an alternative to more tedious procedures for preparation of cereal prolamins.
使用70%乙醇水溶液从谷物面粉中简单提取醇溶蛋白会导致脂质和其他次生产物的共提取。用过量纯乙醇处理粗提取物可去除大部分这些化合物。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）比较时，乙醇沉淀后获得的醇溶蛋白提取物与更复杂的多阶段选择性提取产物几乎没有差异。使用大鼠肝脏溶酶体试验对来自乳糜泻毒性谷物、非乳糜泻毒性谷物和燕麦的乙醇沉淀醇溶蛋白的酶消化物进行了毒性测试。乙醇沉淀提取物中的醇溶蛋白比仅用70%乙醇提取的醇溶蛋白具有更高的活性。由于乙醇沉淀法简单且能提供纯度足以进行进一步评估的醇溶蛋白提取物，因此该方法已被用作制备谷物醇溶蛋白的更繁琐方法的替代方法。
