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=0.998 9）。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。
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
. 高效液相色谱法测定乳及乳制品中唾液酸含量方法研究[J]. 中国乳业, 2023
, 0(11)
: 91
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DOI: 10.12377/1671-4393.23.11.17
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,5 μm) 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.
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