A Novel and Reliable Spectrophotometric Method for Determination of Lactoperoxidase Activity in Raw Milk  

作　　者：Jiang Zhan-mei Che Hong-xia Yang Nan Yuan Xiang-ying Li Xue-yan Zhang Xin-liu 

机构地区：[1]Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China

出　　处：《Journal of Northeast Agricultural University(English Edition)》2017年第2期66-73,共8页东北农业大学学报（英文版）

基　　金：Supported by Project for Research and Development of Harbin Aapplication Technology(2016RAQXJ046);the National "Twelfth Five-year" Plan for Science and Technology Support Program of China(2013BAD18B06)

摘　　要：With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The method was based on the enzymatic reaction principle, where hydrogen peroxide oxidated TMB in the presence of LP. The optimized conditions of this assay system were obtained, consisting of 20 mmol · L-1 TMB solution, 0.6 mmol · L-1 hydrogen peroxide and 0.1 mol · L-1 Citric Acid（CA）/0.2 mol · L-1 disodium hydrogen phosphate（Na P） buffer（pH 4.8）. TMB detection method was applied to the analysis of LP in milk samples with a practical working concentration range from 2 to 14 mg · L-1. The intra-and inter-batch variation coefficients were all below 5%, indicating a good repeatability. Confirmation test between TMB method and 2, 2-azinobi（3-ethylbenzothiazoline-6-sulphonate） diammonium salt（ABTS） method was carried out, and the results of TMB assay were in accordance with that of ABTS method.With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The method was based on the enzymatic reaction principle, where hydrogen peroxide oxidated TMB in the presence of LP. The optimized conditions of this assay system were obtained, consisting of 20 mmol · L-1 TMB solution, 0.6 mmol · L-1 hydrogen peroxide and 0.1 mol · L-1 Citric Acid（CA）/0.2 mol · L-1 disodium hydrogen phosphate（Na P） buffer（pH 4.8）. TMB detection method was applied to the analysis of LP in milk samples with a practical working concentration range from 2 to 14 mg · L-1. The intra-and inter-batch variation coefficients were all below 5%, indicating a good repeatability. Confirmation test between TMB method and 2, 2-azinobi（3-ethylbenzothiazoline-6-sulphonate） diammonium salt（ABTS） method was carried out, and the results of TMB assay were in accordance with that of ABTS method.

关 键 词：lactoperoxidase activity 3 3＇5 5＇-tetramethylbenzidine （TMB） detection condition raw milk 

分 类 号：S823[农业科学—畜牧学]