水牛乳中蛋白质基于RP-HPLC指纹图谱的建立及在乳源分析中的应用  

Establishment of RP-HPLC fingerprint of buffalo milk protein and its application in milk source analysis

作　　者：梁政洋 黄丽 李玲 曾庆坤 赵宏涛 LIANG Zhengyang;HUANG Li;LI Ling;ZENG Qingkun;ZHAO Hongtao(Guangxi Buffalo Research Institute,Nanning 530001;Guangxi Huaxu Buffalo Breeding Co.,LTD.,Nanning 530001)

机构地区：[1]广西壮族自治区水牛研究所,南宁530001 [2]广西华胥水牛繁育有限公司,南宁530001

出　　处：《食品科技》2019年第8期329-334,共6页Food Science and Technology

基　　金：广西重点研发计划项目(桂科AB1850017，桂科AB1850033);南宁市重点研发计划项目(20173162-2)

摘　　要：以水牛的乳蛋白多态性规律,κ-CN/A、αs2-CN、κ-CN/B、αs1-CN、β-CN、β-LgB、α-La在液相色谱中的分离特征和峰面积作为评价指标,对不同来源的水牛乳和荷斯坦牛乳建立指纹图谱快速区分鉴别水牛乳掺假。选择ZORBAX300SB-C8(4.6mm×150mm,3.5μm)色谱柱;检测波长为215nm;柱温:45℃;流速:0.5mL/min;流动相:0.1%TFA水溶液:0.1%TFA乙腈进行梯度洗脱。结果:在精密度、重复性和稳定性实验中,分离出的κ-CN/A、αs2-CN、κ-CN/B、αs1-CN、β-CN、β-LgB、α-La峰面积相对标准偏差(RSD%)<5%;根据水牛与荷斯坦蛋白多态性的差异,对乳蛋白进行分离后建立指纹图谱,通过相似性计算水牛乳和荷斯坦牛乳样品间的相似性均大于0.95,相关性高。通过指纹图谱对比发现特征峰,作为掺假标记,当荷斯坦牛乳掺入2%时可被检测到,在掺入5%~40%荷斯坦牛乳的范围内,相关系数大于0.99。该方法具有良好的重现性和专属性,能够得到样品间具有代表性的指纹图谱,能快速稳定地分析水牛乳中是否掺入荷斯坦奶源。In order to investigate the feasibility of RP-HPLC method for protein separation in buffalo milk. According to the milk protein polymorphisms of Guangxi Chinese buffalo,a fingerprint was established to quickly distinguish the adulteration of buffalo milk.The chromatographic condititions were follows- ZORBAX SB-300 C8 (4.6 mm×150 mm,3.5 μm) was choosed as chromatographic column,the detection wavelength at 215 nm;column temperature at 45 ℃;the flow velocity of 0.5 mL/min;the mobile phase was the mixture of 0.1% TFA in acetonitri:0.1% TFA in water with gradient elution.Result:the peak area relative standard deviation (RSD%) of κ-CN/A,αs2-CN,κ-CN/B,αs1-CN,β-CN,β-LgB,α-La were less than 5% in precision test,the stability test and the repeatability test.Based on the difference of buffalo milk and Holstein milk protein polymorphisms separated by HPLC,establish fingerprint.Both buffalo milk and Holstein milk,similarity between samples great than 0.95.Found the characteristic peak as a distinction mark,when Holstein milk mixed with 2% can be detected.In the range of 5%~40% mixed Holstein milk, the correlation coefficient were all great than 0.99.This method has good reproducibility and specificity, can obtain representative fingerprint between samples,can quickly and stably analyze whether the buffalo milk is adulterated with Holstein milk source.

关 键 词：乳蛋白 高效液相色谱 指纹图谱技术 乳源鉴别 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]