利用氨基酸分析仪对酱油中皮革水解蛋白标志物羟脯氨酸的鉴定

Identification of Hydrolyzed Protein Reagent Protein-Hydroxyproline in Soy Sauce Using Amino Acid Analyzer

[目的]建立一种利用氨基酸分析仪分析鉴定酱油中皮革水解蛋白标志物羟脯氨酸的方法。[方法]选用4-羟基-α-吡咯甲酸为参考物质,利用氨基酸自动分析仪法和紫外分光光度计法对酱油中L-羟脯氨酸的含量进行测定,比较这2种检测方法测定羟脯氨酸的优劣点。[结果]紫外分光光度法在羟脯氨酸的质量浓度为1~6 mg/L与吸光度之间线性关系良好(R 2=0.9980),RSD为0.91%,分别添加0.5、2.0、6.0 mg/L羟脯氨酸标准溶液的加标回收率为90.0%~93.8%。氨基酸分析仪法在羟脯氨酸质量浓度为0.5~30.0 mg/L与峰面积之间线性关系良好(R 2=0.9993),RSD为0.75%,分别添加0.5、10.0、20.0 mg/L羟脯氨酸标准溶液的加标回收率为96.8%~99.7%。[结论]氨基酸分析仪法自动化程度好,重现性和准确度较好,适用于酱油中是否添加动物皮革水解蛋白的鉴定和检测,为食品安全监管提供可靠的技术支持。

[Objective]To set a method for the identification of the hydrolyzed protein marker-hydroxyproline in soy sauce using an amino acid analyzer.[Method]Using 4-hydroxy-α-pyrrolecarboxylic acid as the reference substance,the content of L-hydroxyproline in soy sauce was determined by the automatic amino acid analyzer method and the ultraviolet spectrophotometer method,and the advantages and disadvantages of the two detection methods for the determination of hydroxyproline were compared.[Result]The UV spectrophotometry showed a good linear relationship between the mass concentration of hydroxyproline 1-6 mg/L and the absorbance(R 2=0.9980),the RSD was 0.91%,and the recoveries of standard addition of 0.5,2.0,6.0 mg/L hydroxyproline standard solution were 90.0%-93.8%.Amino acid analyzer method had a good linear relationship between the mass concentration of hydroxyproline 0.5-30.0 mg/L and the peak area(R 2=0.9993),the RSD was 0.75%,the spiked recovery rates of 0.5,10.0,and 20.0 mg/L hydroxyproline standard solutions were 96.8%-99.7%.[Conclusion]The amino acid analyzer method has good automation,good reproducibility and accuracy.It is suitable for the identification and detection of whether to add animal leather hydrolyzed protein in soy sauce,and provides reliable technical support for food safety supervision.