改良考马斯亮蓝试剂法测定壳聚糖及其衍生物中蛋白质含量  

Determination of Protein Content in Chitosan and Its Derivatives by Modified Coomassie Brilliant Blue Reagent Method

作　　者：姜惠萍 刘万顺[2] 宋福来 马海楠 韩宝芹[2] JIANG Huiping;LIU Wanshun;SONG Fulai;MA Hainan;HAN Baoqin(Qingdao Biotemed Biomaterials Co.,Ltd.,Qingdao 266104,China;College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China)

机构地区：[1]青岛博益特生物材料股份有限公司,山东青岛266104 [2]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《现代食品》2022年第17期173-178,共6页Modern Food

基　　金：山东省重点研发计划(重大科技创新工程)“新型海洋生物医用材料研发与制备”(2021CXGC010502)。

摘　　要：经典的考马斯亮蓝试剂测定壳聚糖及其衍生物中的可溶性蛋白含量时,能与氨基多糖样品本身形成絮状沉淀,干扰了蛋白质测定结果的准确度。本文探讨经改良的考马斯亮蓝试剂测定壳聚糖及其衍生物中可溶性蛋白质含量的精确性。选用一定量的盐酸替代磷酸配制考马斯亮蓝试剂,对壳聚糖及其衍生物中蛋白质含量进行测定。结果表明,新配方的考马斯亮蓝试剂与蛋白结合后,染料的最大吸收峰位置发生变化,吸光度与蛋白含量间具有良好的线性关系(R^(2)=0.994 1),回收率为118.74%,最小检出量为1μg·mL。采用改良后的考马斯亮蓝试剂法测定壳聚糖及其衍生物中的微量蛋白质含量准确可靠。When the classical coomassie brilliant blue reagent is used to measure the soluble protein content in chitosan and its derivatives, it can form a flocculent precipitate with the aminopolysaccharide sample itself, which interferes with the accuracy of the protein determination result. This paper discusses the accuracy of the modified coomassie brilliant blue reagent for the determination of soluble protein content in chitosan and its derivatives. A certain amount of hydrochloric acid was used instead of phosphoric acid to prepare coomassie brilliant blue reagent, and the protein content in chitosan and its derivatives was determined. The test results showed that after the new formulation of coomassie brilliant blue reagent was combined with the protein, the position of the maximum absorption peak of the dye changed, and there was a good linear relationship between the absorbance and the protein content(R^(2)=0.994 1), and the recovery was 118.74%, and the minimum detection amount was 1 μg·mL. The modified coomassie brilliant blue reagent method was used to determine the content of trace protein in chitosan and its derivatives accurately and reliably.

关 键 词：考马斯亮蓝 壳聚糖 蛋白质 含量测定 

分 类 号：TS201.2[轻工技术与工程—食品科学]