三文鱼中晚期糖基化终末产物含量测定及产地特征因子分析

Determination of Advanced Glycation End Products in Salmon and Exploration of Characteristic Indicators to Identify Its Geographical Origin

建立超高效液相色谱-串联三重四极杆质谱同时检测三文鱼中羧甲基赖氨酸(N-carboxymethyllysine,CML)和羧乙基赖氨酸(N-carboxyethyllysine,CEL)的分析方法。色谱柱为ACQUITY UPLC HSS T3柱(2.1 mm×150 mm,1.8μm),流动相A为100%乙腈、B为0.1%甲酸,电喷雾离子源正离子模式扫描,多反应监测模式定性分析,同位素内标稀释法定量;样品经脱脂、还原、水解和Oasis MCX固相萃取柱净化等步骤后上机分析;方法平均回收率为90%~95%,精密度相对标准偏差小于5%,CEL检出限为0.0327 ng/g,定量限为0.1422 ng/g。采用该法对来自挪威、智利和法罗群岛的21份三文鱼样品进行含量测定,并对所获数据进行主成分分析。结果表明,即使是相同品种的三文鱼在不同国家和不同个体之间CML和CEL含量存在很大的生物差异性,并且发现CEL有望成为鉴别挪威三文鱼和智利、法罗群岛三文鱼的产地特征因子。

A method for simultaneous determination of carboxymethyllysine(N-carboxymethyllysine,CML)and carboxyethyllysine(N-carboxyethyllysine,CEL)in salmon by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was developed in this paper.The chromatographic separation was performed on an ACQUITY UPLC HSS T3 column using a mobile phase composed of 100%acetonitrile and 0.1%formic acid,and the separated analytes were detected by electrospray ionization in the positive ion(ESI+)mode under multiple reaction monitoring(MRM)and quantitated by the isotope internal standard dilution method.Samples were degreased,reduced,and hydrolyzed and the hydrolysate was purified by solid phase extraction on an Oasis MCX column before being injected into the instrument.The average recoveries of the method were 90%–95%,the precision expressed as relative standard deviation(RSD)was less than 5%,the limit of detection(LOD)was 0.0327 ng/g,and the limit of quantitation(LOQ)was 0.1422 ng/g.The method was used to detect 21 salmon samples from Norway,Chile and the Faroe Islands,and the obtained data were analyzed by principal component analysis(PCA).The results showed that there were significant differences in CML and CEL contents between different countries and individuals of the same species.This study revealed CEL to be a potential characteristic indicator to distinguish Norway salmon from those from Chilean and the Faroe Islands.