UPLC快速测定可溶性肽或蛋白中谷氨酰胺含量  

Quick determination of glutamine in soluble protein or peptide by UPLC

作　　者：张海华[1] 陶冠军[2] 周惠明[1] 

机构地区：[1]江南大学食品学院,江苏无锡214122 [2]江南大学食品科学与技术国家重点实验室,江苏无锡214122

出　　处：《食品工业科技》2012年第1期338-339,365,共3页Science and Technology of Food Industry

基　　金：国家高科技研究发展计划863项目(2008AA10Z312);江南大学博士基金项目(2009)

摘　　要：建立了UPLC测定谷氨酰胺的方法。采用BTI([bis(trifluoroacetoxy)iodo]benzene,BTI)衍生化肽或蛋白中非氮端的谷氨酰胺(Glutamine,Gln),衍生产物L-2,4-二氨基丁酸(L-2,4-Diaminobutyric acid,DABA)经2,4-二硝基氟苯(2,4-Dinitrofluorobenzene,DNFB)紫外显色后,用超高效液相色谱仪(UPLC)在355nm下进行检测测定。结果表明,DABA的最低检出限为0.2ng/mL,线性范围为1～10μg/mL。经Ala-Gln、β-乳球蛋白、α-乳清蛋白和溶菌酶四种蛋白标品检验,测得标样中Gln的回收率为93%±2%～98%±3%。所用方法具有检出限低、样品用量少、高效快速等特点。A method of UPLC determining glutamine was developed.Glutamine in Non-nitrogen terminal of soluble protein or peptides were accordingly converted into stable L-2,4-diaminobutyric acid（DABA） in the presence ofbis（trifluoroacetoxy）iodobenzene（BTI）.Thereafter,that DABA reacted with 2,4-Dinitrofluorobenzene（DNFB） gave rise to a final product which was detected at 355nm by UV detector accessory for UPLC.The results showed that the lowest detection limit of DABA was 0.2ng/mL,and the linear region was 1～10μg/mL.The method recovery was tested with four standards including Ala-Gln,β-lactoglobulin,α-albumin and Lysozyme,a recovery range of 93%±2%～98%±3% was obtained.In general,the method developed has advantages of quickness,lower detection limit and less sample.

关 键 词：谷氨酰胺 超高效液相色谱 L-2 4-二氨基丁酸 2 4-二硝基氟苯 

分 类 号：TS207.3[轻工技术与工程—食品科学]