摘要
探寻一种准确测定牛初乳中IgG含量的前处理方法及液相色谱分离条件,选用超声波、生理盐水、盐酸和乳酸作为去除乳脂和酪蛋白沉淀的前处理条件;选用反相键合色谱柱ZORBAX 300SB-C18和ZORBAX Poroshell 300SB-C18,流动相为0.02mol/L PBS与乙腈作为色谱分离条件对初乳清进行分离;并测定其精密度和回收率。试验结果表明:选用超声波40kHZ、45min时,牛初乳中IgG含量达到最高,比没有经过超声处理的IgG含量提高近两倍;选用乳清与生理盐水的稀释比例为1:7时,牛初乳中IgG含量达到最高;选用乳酸调pH值至4.6时,可以更好的去除酪蛋白沉淀;选用反相键合色谱柱ZORBAX 300SB-C18,流动相0.02mol/L PBS(pH6.8)与乙腈的比例为95:5、流速为0.2mL/min时,初乳各蛋白组分分离效果达到最佳。所测定的精密度和回收率分别为0.11%和99.96%。
The way of preceding disposition and the separate condition of HPLC of the content IgG in bovine colostrums were explored. The ultrasonic, brine, hydrochloric acid and lactic acid were selected as the condition of preceding disposition; the contrary bond ehromatograln column ZORBAX 300SB-C18 and ZORBAX Poroshell 300SB-C18, the different proportion of 0.02 mol/L PBS and Ethioninein was selected as the separate condition of chromatograph; and the exactitude and callback ratio were also determined. The result were as follows： the content of IgG is the highest when the ultrasonic is 40 kHZ,45 rain and the diluted proportion between whey and brine is 1：7;The casein deposition is wiped off when the lactic acid and pH 4.6 are selected. The separate effect was very good when the contrary bond chromatogram column ZORBAX 300SB-C18, the proportion of 0.02 mol/L PBS （95%） and Ethioninein （5%）, the flow velocity of 0.2 mL/min were selected. The exactitude and callback ratio is 0.11% and 99.96% respectively.
出处
《乳业科学与技术》
2007年第5期224-229,共6页
JOURNAL OF DAIRY SCIENCE AND TECHNOLOGY
基金
山东省自然科学基金(奶牛热应激的补偿生长机制研究
2004ZX27)
