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目的建立测定营养型固体饮料中维生素B_(12)水平的高效液相色谱测定方法。方法样品经胃蛋白酶和淀粉酶水解后,采用维生素B_(12)免疫亲和柱富集、净化,用5ml甲醇洗脱,氮气吹干定容,再经高效液相色谱法测定,色谱柱为Eclipse XDB-C_(18),流动相有体积分数为0.025%三氟乙酸水溶液和体积分数0.025%三氟乙酸乙腈溶液梯度洗脱,流速为1.0 ml/min,波长为361nm测定。结果维生素B_(12)在21.0～420.0mg/L范围内线性关系良好,回归方程y=0.115x+0.024,相关系数0.9999,回收率为88.5%～94.6%,检出限为0.5ng。结论本研究方法灵敏度高,结果准确,适用于测定营养型固体饮品中维生素B_(12)的水平。
Objective To establish a HPLC method for the determination of vitamin B 12 in nutritional solid beverages. Methods After the sample was hydrolyzed by pepsin and amylase, it was enriched with vitamin B 12 immunoaffinity column and purified by eluting with 5 ml of methanol. The sample was purged with nitrogen and dried, and then determined by high performance liquid chromatography Eclipse XDB-C_ (18), the mobile phase has a volume fraction of 0.025% trifluoroacetic acid aqueous solution and a volume fraction of 0.025% trifluoroacetic acid acetonitrile gradient elution at a flow rate of 1.0 ml / min, a wavelength of 361nm determination. Results The calibration curve of vitamin B 12 was good in the range of 21.0-420.0 mg / L. The regression equation y = 0.115x + 0.024, the correlation coefficient was 0.9999, the recovery was 88.5% -94.6%, and the detection limit was 0.5ng. Conclusion The method of this study has high sensitivity and accurate results and is suitable for the determination of vitamin B 12 in nutritional solid drinks.