乳和乳制品中动物水解蛋白鉴定——比色法测定L（-）-羟脯氨酸含量

摘要

目的：研究乳和乳制品中动物水解蛋白的鉴定方法。通过对L（-）-羟脯氨酸的检测来鉴定是否含有动物水解蛋白。方法：样品经酸水解,释放出羟脯氨酸。经氯胺T氧化,生成含有吡咯环的氧化物。用高氯酸破坏过量的氯胺T。羟脯氨酸氧化物与对二甲氨基苯甲醛反应生成红色化合物,在波长558nm处进行比色测定。结果：动物水解蛋白在110℃条件下水解6h后,能够充分水解为L（-）-羟脯氨酸。以L（-）-羟脯氨酸为标样,比色法测定乳制品中L（-）-羟脯氨酸含量。该方法的加标回收率为99.6%～104.6%,满足分光光度法分析方法的实验需求。%Objective： To establish a colorimetric method to determine L（-）-hroxyproline content for identifying whether hydrolyzed animal protein is present in milk and milk products.Methods： L（-）-Hroxyproline was released from samples treated by acid hydrolysis and oxidized by chloramine T to form oxides containing pyrrole rings.Perchloric acid was then added to damage excess chloramine T.The L（-）-hroxyproline oxides were allowed to react with p-dimethylaminobenzaldehyde to generate red compounds for colorimetric determination at 558 nm wavelength.Results： Hydrolyzed animal protein was completely hydrolyzed into L（-）-hroxyproline after 6 h of hydrolysis at 110 ℃.The stability,reproducibility and spike recovery of the developed method were all consistent with requirements.