液相色谱串联质谱检测牙鲆鱼中的小清蛋白过敏原  

Determination of allergen parvalbumin in Paralichthys olivaceus using HPLC-MS/MS

作　　者：葛敏敏 王建华 林洪[1] GE Minmin;WANG Jianhua;LIN Hong(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China;Inspection and Couarantine Technical Center,Qingdao Customs,Qingdao 266002,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]青岛海关检验检疫技术中心,山东青岛266002

出　　处：《食品与发酵工业》2019年第8期210-214,共5页Food and Fermentation Industries

基　　金：国家自然科学基金(31771892)

摘　　要：以合成肽段ALTDAETK为定量特征肽段,SDFIEEDELK及LFLQNFAFSASAR为定性肽段,建立牙鲆中小清蛋白高效液相色谱-串联质谱的检测方法。同时对蛋白提取液、碘代乙酰胺浓度、酶/蛋白质比例、酶解时间在内的前处理条件和肽段质谱参数条件进行了优化。结果显示,ALTDAETK肽段在0. 005～10 mg/L线性关系良好(R2> 0. 999),小清蛋白定量限2. 74 mg/kg;以大菱鲆为空白基质,重组小清蛋白的平均回收率为95%～102%,相对标准偏差(RSD)小于7%,重复性好。该文首次实现了用HPLC-MS/MS方法精确定量检测牙鲆中主要过敏原PV。A method using high performance chromatography tandem mass spectrometry/mass spectrometry(HPLC-MS/MS)detecting parvalbumin in Paralichthys olivaceus was established.ALTDAETK was used as a quantitative marker peptide,SDFIEEDELK and LFLQNFAFSASAR were used as qualitative marker peptides.The pretreatment conditions including protein extracts,indole-3-acetic acid (IAA) concentration,enzyme/protein ratio,and enzymatic hydrolysis time,as well as the instrumental conditions for mass spectrometry parameters used for three marker peptides were optimized.The results showed that ALTDAETK had a good linear relationship in a range of 0.005- 10 mg/L (R^2>0.999).The quantification limit of parvalbumin was 2.74 mg/kg.Took Psetta maxima as a blank,the average recovery rate of recombinant parvalbumin was 94.8%-102.0%,and the relative standard deviation (RSD) was below 7% with a good repeatability.It was the first time that parvalbumin was detected in Paralichthys olivaceus accurately and quantitatively using HPLC-MS/MS.

关 键 词：牙鲆 小清蛋白 高效液相色谱串联质谱(HPLC-MS/MS) 过敏原 检测 

分 类 号：O657.63[理学—分析化学] TS254.7[理学—化学]