高效液相色谱法测定乳及乳制品中唾液酸含量方法研究  

Determination of salivary acid in milk and milk products by high performance liquid chromatography

作　　者：赵梓棋 李艳红 李慧 卢智华 张红梅 齐琦 毕成名 王丹慧 王斌 白晓玲 张河霞 ZHAO Ziqi;LI Yanhong;LI Hui;LU Zhihua;ZHANG Hongmei;QI Qi;BI Chengming;WANG Danhui;WANG Bin;BAI Xiaoling;ZHANG Hexia(Mengniu Dairy Limited Liability Company,Hohhot Inner Mongolia 011500)

机构地区：[1]蒙牛乳业有限公司,内蒙古自治区呼和浩特市011500

出　　处：《中国乳业》2023年第11期91-99,共9页China Dairy

摘　　要：研究开发了一种以高效液相色谱法(HPLC)为基础的方法,用于检测唾液酸原样及乳制品中N-乙酰神经氨酸的含量。检测样品在经过前期处理后,采用C18色谱柱(250 mm×4.6 mm,5μm),梯度洗脱法中使用0.005 mol/L的硫酸溶液和0.005 mol/L的硫酸甲醇溶液作为流动相。通过单因素和正交实验优化实验条件,当柱温为40℃、流速为0.6 mL/min、流动相pH值为2.0、流动相比例为85∶15(V/V)时,N-乙酰神经氨酸的保留时间为5.69 min,分离效果及精密度较好,在0~3.0 mg/mL的浓度范围内有良好的线性关系(R2=0.9989)。加标回收率在99.2%~103.4%之间,精密度检测(relative standard deviation,RSD)在0.47%~1.26%之间。方法操作简便,结果准确可靠,精密度良好,提高了检验人员的技术操作性,适用于唾液酸原样及乳制品中的N-乙酰神经氨酸含量分析。A high performance liquid chromatography(HPLC)-based method was developed for the determination of N-acetylneuraminic acid added to raw materials and dairy products.After pre-treatment of the test samples,a C18 column(250 mm×4.6 mm,5μm)was used,and 0.005 mol/L sulfuric acid solution and 0.005 mol/L methanol sulfuric acid solution were used as mobile phases in the gradient elution method.The experimental conditions were optimized by one-way and orthogonal experiments,and the retention time of N-acetylneuraminic acid was 5.69 min when the column temperature was 40℃,the flow rate was 0.6 mL/min,the pH value of the mobile phase was 2.0,and the ratio of the mobile phase was 85∶15(V/V),which showed good separation and precision,and a good linearity in the concentration range of 0~3.0 mg/mL(R2=0.9989).The spiked recoveries ranged from 99.2%~103.4%,and the precision tests(relative standard deviation(RSD)ranged from 0.47%to 1.26%.The method is easy to operate,with accurate and reliable results and good precision,which not only improves the technical operation of the inspectors,but also applies to the analysis of N-acetylneuraminic acid content in raw materials and dairy products.

关 键 词：唾液酸 高效液相色谱法 乳制品 检测方法 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程] O657.72[轻工技术与工程—食品科学与工程]