烤羊肉中丙烯酰胺的HPLC测定方法研究及应用

HPLC Determination Method of Acrylamide in Roast Mutton and Its Application

为了考察羊肉在烤制过程中是否产生有害物质丙烯酰胺,并考虑到由于基质的不同,现有食品中丙烯酰胺的HPLC检测方法可能不适用于烤羊肉中丙烯酰胺的检测。因此本文筛选了烤羊肉样品前处理方法(萃取溶剂、蛋白质沉淀剂)及高效液相色谱法(HPLC)色谱条件,同时验证了HPLC法测定烤羊肉中丙烯酰胺的可行性,并用该法检测了烤羊肉中的丙烯酰胺含量。烤羊肉样品中加50%甲醇制备匀浆,用甲醇沉淀蛋白,上清液加正己烷脱脂后,采用真空冷冻离心浓缩法挥干上清液,再用超纯水复溶;采用Target TC C18(150 mm×4.6 mm,5μm)色谱柱,以甲醇∶0.1%甲酸水为流动相梯度洗脱,流速为0.6 mL/min,进样量10μL;实验结果表明,丙烯酰胺在0.1μg/mL-1.6μg/mL范围内线性关系良好,萃取回收率为99.38%-100.40%,加样回收率为97.67%-100.83%,RSD为1.26%-2.66%,显示了良好的准确度及精密度。进一步利用该法检测了不同来源、不同批次烤羊肉中的丙烯酰胺,结果显示,烤羊肉中也存在较高量的丙烯酰胺,其含量在0.63-1.16μg/g之间。该方法简便,快捷,可靠,适用于烤羊肉中丙烯酰胺的检测。

To investigate whether acrylamide is produced during the mutton roasting process,and consider that the HPLC method may not be suitable for testing acrylamide in roast mutton due to the different matrices. Therefore the sample pretreatment conditions(extracting solvent and protein precipitator) and the HPLC conditions were optimized,and meanwhile,the feasibility of determination of acrylamide was also verified,and the content of acrylamide was determined by this method in roast mutton. The samples were homogenated with 50% methanol water,the protein was precipitated by methanol,the grease was removed by hexane,vaporized by vacuum freezing centrifugal concentration,and then dissolved by ultrapure water. The separation was performed on the Target TC C18(150 mm × 4.6 mm,5 μm) column with gradient elution,the mobile phase was 0.1% formic acid water solution and methanol,and the flow rate was set as 0.6 m L/min. The injection volume was set as 10 μL. The linear range of acrylamide was within 0.1 μg/mL-1.6 μg/mL. The extraction recovery was in the range of 99.38%-100.40%,the standard recovery rate was in the range of 97.67%-100.83%,the relative standard deviations was1.26%-2.66%,and good accuracy and precision were shown by them. The established method was used to evaluate acrylamide in different batches of roast mutton,the findings showed that there was a amount of acrylamide in roast mutton between the range of 0.63-1.16 μg/g. This method was simple,fast and reliable,suitable for the analysis and determination of acrylamide in roast mutton.