通过对β-乳球蛋白酶解反应的各个关键条件进行优化,确定了还原烷基化试剂用量为50 μL、还原温度为55℃、胰蛋白酶溶液浓度为1 mg/mL、酶解温度为37℃、酶解时间为16 h、终止酶解试剂为10%甲酸溶液,在该条件下特征多肽酶解较为完全;并对色谱检测条件进行优化,利用同位素稀释质谱法准确定量乳品中β-乳球蛋白,RSD<7%,可为乳品中过敏原准确定量提供一定的参考。

Through optimizing key conditions for the enzyme lysis reaction of β-lactoglobulin,it is determined that the dosage of reducing alkylation reagent is 50 μL,the reduction temperature is at 55℃,the concentration of trypsin solution is 1 mg·mL^-1,enzymatic lysis temperature is at 37℃,the enzymatic lysis lasts for 16 h,and 10% formic acid solution is used as the terminating enzyme lysis reagent.Under these conditions,the enzyme lysis of characteristic peptide can be relatively complete.β-Lactoglobulin in dairy products is accurately quantified by isotope dilution mass spectrometry,with an RSD less than 7%,which can provide reference for the accurate quantification of allergens in dairy products.

徐增益(1995-),女,硕士生,主要从事稳定同位素分析检测方面的研究