免疫亲和柱—超高效液相色谱法测定维生素饮料中维生素B12

摘要

采用免疫亲和柱—超高效色谱法建立维生素饮料中维生素B12的测定方法,样品经免疫亲和柱富集、净化后,用3mL甲醇以流速为1滴/秒的速度洗脱,氮气吹干后溶解再经超高效液相色谱法测定.色谱柱为HSS T3,柱温35C,流动相为乙腈-0.1％甲酸溶液梯度洗脱,流速为0.6mL/min,波长为360nm测定.结果表明,维生素B12在浓度为0.4 ～ 10μg/L内线性关系良好,回归方程为Y=4 040X-364,相关系数为0.997 00,回收率为94.68％,检出限为0.1 μg/L.本方法具有操作简单、灵敏度高、结果准确、检测效率高等优点,满足于成分复杂、维生素含量低的维生素饮料中B12的测定要求.%Super efficient immune affinity column chromatography was adopted to establish the method for determination of the vitamin B12 in vitamin drink,sample after immune affinity column,purification,with 3 mL methanol elution at a speed of flow rate of 1 drop/See,nitrogen blow dry after dissolved by Ultra High Performance Liquid Chromatography (UPLC) method.Chromatographic column for HSS T3,column temperature 35 ℃,mobile phase gradient elution of acetonitrile-0.1％ formic acid solution,flow rate of 0.6 mL/min,the wavelength of 360 nm.Results showed that the concentration of vitamin B12 in the 0.4 ～ 10 μg/L range inside sex was good,the regression equation of y =4 040 x-4 040,the correlation coefficient was 0.997 00,recovery rate was 94.68％,the detection limit was 0.1 μg/L This method has the advantages of simple operation,high sensitivity,accurate,high detection efficiency,and is suitable for the determination of vitamin B12 in drinks with complex composition and low vitamin content.