基于牛乳非蛋白生物标志物定量检测山羊乳中掺加的牛乳  

Quantitative Detection of Cow Milk Adulteration in Goat Milk Based on a Non-Protein Biomarker Present in Cow Milk

作　　者：孙金芝 付路静 岳田利[1] 李婧妍[1] 郭春锋[1] SUN Jinzhi;FU Lujing;YUE Tianli;LI Jingyan;GUO Chunfeng(College of Food Science and Engineering,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100

出　　处：《食品科学》2021年第8期288-293,共6页Food Science

基　　金：陕西省重点研发计划一般项目(2019NY-113;2020NY-184)。

摘　　要：为克服酶联免疫吸附法无法检测羊乳中掺加的经过热处理的牛乳的局限,本研究首先通过气相色谱-质谱法发现牛乳区别于山羊乳的关键非蛋白生物标志物为N-乙酰氨基葡萄糖,然后基于该标志物开发基于高效液相色谱手段定量检测山羊乳中掺加的牛乳的方法。样品在70℃经1-苯基-3-甲基-5-吡唑啉酮衍生60 min后,在高效液相色谱仪上使用反相C18色谱柱对目标物进行分离,在245 nm紫外波长下对其进行检测,根据N-乙酰氨基葡萄糖含量推算山羊乳中掺加的牛乳量。该方法的牛乳掺加量检出限和定量限分别为0.3%和1.0%,在1%~100%的牛乳掺加范围内线性良好,相关系数为0.9994,该方法的加标回收率为100.4%~105.1%,在10、50 mg/L和100 mg/L加标质量浓度下,日内和日间精密度分别为1.8%和2.0%、1.7%和3.7%、2.6%和2.7%。该方法具有较高的灵敏度、准确度和精密度,可用于山羊乳中掺加牛乳的定量分析。Enzyme-linked immunosorbent assay cannot detect the adulteration of heat-treated cow milk in goat milk.In this study,N-acetylglucosamine was revealed by gas chromatography-mass spectrometry to be a non-protein biomarker present in cow milk that discriminates it from goat milk,and a high performance liquid chromatographic method was developed for quantitative detection of cow milk adulteration in goat milk.Samples were derivatized with 1-phenyl-3-methyl-5-pyrazolone at 70℃for 60 min,followed by chromatographic separation of the marker on a reversed-phase C18 column and detection at 245 nm.The amount of cow milk added to goat milk was calculated according to the N-acetylglucosamine content in the samples.The limit of detection and limit of quantification of the method were 0.3%and 1.0%,respectively.The method showed good linearity in the range of 1%–100%cow milk addition with correlation coefficient of 0.9994.The recoveries of spiked samples were 100.4%–105.1%,and the intra-day and inter-day precisions(relative standard deviations)at spiked concentrations of 10,50 and 100 mg/L were 1.8%and 2.0%,1.7%and 3.7%,2.6%and 2.7%,respectively.This method proved to be highly sensitive,accurate and precise,and therefore could be suitable for quantitative analysis of cow milk adulteration in goat milk.

关 键 词：山羊乳 牛乳 掺假 N-乙酰氨基葡萄糖 高效液相色谱法 

分 类 号：Q532[生物学—生物化学]