LC-MS/MS分析结合优化提取工艺探究啤酒中麸质蛋白  

Analysis of the Gluten Map in Beers by LC-MS/MS Combined with Optimized Extraction Protocol

作　　者：于欣禾 李明慧[1] 孙珍 YU Xin-he;LI Ming-hui;SUN Zhen(School of Biological Engineering,Dalian Polytechnic University,Dalian 116034,China;School of Chemistry,Central China Normal University,Wuhan 430079,China)

机构地区：[1]大连工业大学生物学院,辽宁大连116034 [2]华中师范大学化学学院,湖北武汉430079

出　　处：《质谱学报》2022年第2期242-251,I0006,共11页Journal of Chinese Mass Spectrometry Society

摘　　要：大多数啤酒蛋白来自大麦和小麦,它们含有与乳糜泻疾病有关的富含脯氨酸的麸质蛋白成分。本研究选择0.1 mol/L醋酸铵甲醇和丙酮作为优化溶液,分别提取啤酒样品中醇不溶和醇溶的麸质蛋白组分,最大限度地获取麸质蛋白信息。采用优化的麸质蛋白提取工艺,结合液相色谱-质谱法(LC-MS/MS)分析8种市售啤酒的麸质蛋白。结果表明,蛋白组分和多肽组分的结合可以显著提高麸质蛋白的鉴别率。在包括无麸质啤酒在内的所有类型啤酒中,2种蛋白和多肽组分中均能鉴定出麸质蛋白。质谱法可作为酶联免疫吸附测定法的补充,以确定无麸质啤酒的安全性。Gluten,which accounts for 70%-80%of the total protein content of wheat,is the main family of storage proteins found in the starchy endosperm of Pooideae subfamily,such as wheat,barley,rye and oats.As described in previous research,it could be divided into two fractions based on their different solubility in aqueous alcohol.Alcohol-soluble proteins of cereal grains that are rich in proline(Pro)and glutamine(Gln)are termed prolamins.The word of prolamins is a collective name of alcohol-soluble,water-insoluble storage proteins from wheat(gliadin),barley(hordein),rye(secalin),and oat(avenin).The alcohol-insoluble fraction of wheat gluten is called glutenin and the corresponding fractions from other species glutenins.Celiac disease(CD)is defined as a chronic T-cell mediated enteropathy caused by exposure to dietary gluten in genetically predisposed individuals,which account for approximately 1%of the global population.Clinical symptoms of CD can vary of patients and may include intestinal symptoms of diarrhea or steatorrhea,as well as extraintestinal symptoms,such as bone pain or osteoporosis.The only effective treatment for individuals with CD is lifelong adherence to strict gluten-free diet.Beer is a popular alcohol beverage fermented with cereals such as barley and wheat which contains antigenic proteins of CD.Gluten-free beer is also specially provided by manufacturers for CD sufferers.During the whole process of beer manufacturing,hordeins and other storage proteins are degraded by proteases which are activated during malting.The high temperature during mashing may also cause the modification of gluten proteins.So the gluten proteins can be degraded to produce soluble polypeptides and free amino acids,and the epitopes used for the ELISA protocol are also damaged,which hinders the accurate detection of gluten proteins by ELISA protocols.In this work,0.1 mol/L ammonium acetate in methanol and acetone was selected as the optimized solution to individually obtain alcohol-insoluble and alcohol-soluble fractions of glu

关 键 词：液相色谱-串联质谱(LC-MS/MS) 麸质蛋白 啤酒 乳糜泻疾病 提取 去酰胺化 

分 类 号：O657.63[理学—分析化学]