高效阴离子交换色谱法测定婴幼儿配方奶粉中酵母β-葡聚糖

Determination of yeast β-glucan in infant formula by high performance anion exchange chromatography

建立一种测定婴幼儿配方奶粉中酵母β-葡聚糖的分析方法。奶粉样品经蛋白酶酶解去除蛋白质,脂肪酶酶解去除脂肪,葡聚糖酶酶解β-葡聚糖产生葡萄糖,经高效阴离子交换色谱-脉冲安培检测,计算β-葡聚糖含量。结果表明,最优酶解条件为:酶解温度50℃、酶解时间2.0 h、溶壁酶添加量500μL。采用PA10阴离子交换柱进行分离葡萄糖,葡萄糖在0.10~50.0 mg/L范围内线性良好(R;>0.999 5),添加水平在10.0~50.0 mg/100 g范围的回收率为96.9%~102.1%,精密度试验结果相对标准偏差为2.43%(n=5),方法检出限为3.0 mg/100 g,定量限为10.0 mg/100 g。该方法灵敏、准确,可用于婴幼儿配方奶粉中酵母β-葡聚糖的测定。

An analytical method for the determination of yeast β-glucan in infant formula was established. The infant formula sample was subjected to protease hydrolysis to remove protein, lipase hydrolysis to remove fat, glucanase enzymatic hydrolysis β-glucan to produce glucose, which was subjected to high-efficiency anion exchange chromatography-pulse amperometric detection, and then the content of β-glucan was calculated. The results showed that the optimal enzymolysis conditions were as follows: enzymolysis temperature 50 ℃, enzymolysis time 2.0 h, lysozyme addition 500 μl.Glucose was separated by PA10 anion exchange column, the linear relationship of glucose was good in the range of 0.10-50.0 mg/L(R;>0.999 5), the range of recovery rate was 96.9%-102.1% at the addition level of 10.0-50.0 mg/100 g, the relative standard deviation of precision tests results was2.43%(n=5), the detection limit of the method was 3.0 mg/100 g, and the quantification limit was 10.0 mg/100 g. The method was sensitive and accurate,which could be used for the determination of yeast β-glucan in infant formula milk.