摘要
为了建立快速准确检测猪乳中溶菌酶、乳铁蛋白、α-乳白蛋白、血清白蛋白和β-乳球蛋白高效液相色谱方法,采用0.1%三氟乙酸水溶液为流动相A,0.1%三氟乙酸乙腈溶液为流动相B进行程序洗脱,用Xbridge^（TM） Sneild RP C18色谱柱对α-乳白蛋白、β-乳球蛋白和溶菌酶进行分离检测,用Supelcosil C18色谱柱对乳铁蛋白和血清白蛋白进行分离检测。结果显示：5种乳清蛋白分离良好,在25-1 000 mg·L^（-1）范围内呈良好线性,标准曲线回归系数均大于0.99,RSD小于5%,回收率在91%-97%,溶菌酶、乳铁蛋白、α-乳白蛋白、血清白蛋白和β-乳球蛋白的检测限分别为0.2、4.0、0.6、0.3和1.0 mg·L^（-1）;对猪乳中乳清蛋白进行测定显示,β-乳球蛋白、溶菌酶、乳铁蛋白和血清白蛋白含量在母猪分娩后16 d内逐渐降低,其含量均在母猪分娩后第1天最高,分别为17.2、0.224、2.0和15.6 g·L^（-1）,在第16天含量最低,分别为5.6、0.092、0.5和3.6 g·L^（-1）;α-乳白蛋白含量在母猪分娩后逐渐升高,在第7天时含量达到最高为3.3 g·L^（-1）,随后降低,保持在3.1 g·L^（-1）。结果表明：建立的乳清蛋白高效液相色谱法,能够快速准确检测猪乳中5种乳清蛋白的含量。
This study was to establish a rapid and precise method with high-performance liquid chromatography（ HPLC） for determination of lysozyme,lactoferrin,α-lactalbumin,serum albumin and β-lactoglobulin. Chromatographic columns such as Supelcosil C18（ 250 mm ×4. 6 mm,5 μm） and Xbridge^（TM） Sneild RP C18（ 150 mm × 4. 6 mm,5 μm） were used separately. The chromatographic separation was performed using the following eluents： solvent A,0. 1% trifluoroacetic acid（ TFA） in a solution of acetonitrile（ ACN）-H2O（ 5： 95）; and solvent B,0. 1% TFA in ACN-H2O（ 95： 5）. The results showed that the five proteins could be well separated. The calibration curves were linear in the ranges of 25-1 000 mg·L^（-1） with all the correlation coefficients higher than 0. 99 and the relative standard deviations lower than5%,the recoveries of the five proteins were in the ranges of 91%-97%. The detection limits of lysozyme,lactoferrin,α-lactalbumin,serum albumin and β-lactoglobulin were 0. 2,4. 0,0. 6,0. 3 and 1. 0 mg·L^（-1）,respectively. The whey proteins in sow milk were determined using this method. The results showed that the contents of β-lactoglobulin,lysozyme,lactoferrin and serum albumin decreased gradually within 16 days after the parturition,the contents of them were the highest on the first day and were 17. 2,0. 224,2. 0 and 15. 6 g·L^（-1）,respectively. The contents of β-lactoglobulin,lysozyme,lactoferrin and serum albumin were the lowest on the sixteenth day after parturition and were 5. 6,0. 092,0. 5 and 3. 6 g·L^（-1）,respectively. The content of α-lactalbumin increased gradually after parturition,with the highest value of about 3. 3 g·L^（-1） on the seventh day; however,the content of α-lactalbumin decreased to and remained at 3. 1 g·L^（-1）. In conclusion,the HPLC method here to determine whey proteins could effectively detect the contents of whey proteins in sow milk.
作者
杨怀荣
田时
赵道远
胡平
王珏
汪晶
吕波
YANG Huairong;TIAN Shiyi;ZHAO Daoyuan;HU Ping;WANG Jue;Wang Jing;LU Bo(National Center for International Research on Animal Gut Nutrition, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China;College of Science, Nanjing Agricultural University, Nanjing 210095, China)
出处
《畜牧与兽医》
北大核心
2018年第6期19-24,共6页
Animal Husbandry & Veterinary Medicine
基金
中央高校自主创新项目(KYZ201722)
