凝胶电泳分离-液相色谱-质谱法分析林蛙油及其伪品中的特异性蛋白

摘要

本研究应用凝胶电泳分离、蛋白酶酶解、纳升液相色谱分离、串联质谱检测、数据库检索等技术,建立了林蛙油及伪品青蛙输卵管、牛蛙卵巢、牛蛙输卵管和明太鱼输卵管中特异性蛋白的鉴定方法.根据定性肽段数量、肽段的响应高低,蛋白在电泳图谱中条带位置的特异性等因素综合判断,筛选出林蛙油及其相关伪品的特异性蛋白,通过对特异性蛋白的分析测定,判断样品的真伪.采用该方法对市场上的4种林蛙油产品进行检测,通过分析特异性蛋白,发现所检测的样品均为林蛙油伪品.研究结果表明,利用凝胶电泳分离液相色谱质谱法构建的特异性蛋白识别技术可以有效检测林蛙油产品的真伪.%A method for the identification of Oviductus ranae samples was established by gel electrophoresis combined with nano liquid chromatography-mass spectrometry(LC/MS).The samples including Oviductus ranae, Rana catesbeiana fallopian tube, Rana catesbeiana ovary, Rana nigromaculata ovary and walleye pollack fallopian tube were extracted by using the TCA acetone precipitation method.Comprehensive application of gel electrophoresis, enzymatic hydrolysis, nano liquid chromatography, series spectrum detection and database retrieval technology were carried on the analysis to the proteins of Oviductus ranae.At the same time, the proteins of market samples were analyzed using the same method and compared with the proteins in Oviductus ranae.The results show that the specific protein bands distribution of Oviductus ranae and the counterfeits are different, so the authenticity of Oviductus ranae samples is able to be detected by the comparation of the specific protein bands.The method is reliable for the recognition of specific proteins in Oviductus ranae.