超高效液相色谱法同时测定配方乳粉中的烟酸、烟酰胺和VB6

Simultaneous Determination of Niacin,Nicotinamide,and Vitamin B_(6) in Formula Milk Powder by Ultra Performance Liquid Chromatography

建立一种能够同时测定配方乳粉中烟酸、烟酰胺和VB6的超高效液相色谱(ultra performance liquid chromatography,UPLC)分析方法。以10 mmol/L乙酸铵溶液(含0.1%甲酸)为流动相A,甲醇为流动相B进行梯度洗脱,将Poroshell 120 EC-C18与ACQUITY UPLC?HSS T3色谱柱串联分离目标物,经紫外检测器和荧光检测器检测,用外标法定量。结果表明:烟酸质量浓度为0.0~0.8μg/mL、烟酰胺质量浓度为0.0~20.0μg/mL、吡哆胺质量浓度为0.0~0.2μg/mL、吡哆醛质量浓度为0.0~0.6μg/mL,以及吡哆醇质量浓度为0.0~4.0μg/mL,5种化合物标准曲线线性良好,相关系数均大于0.999,检出限和定量限均符合要求;该方法测定的平均回收率为80.5%~104.1%,相对标准偏差为0.37%~2.99%。该分析方法简便、准确,可进行配方乳粉中烟酸、烟酰胺和VB6的批量测定。

To establish an ultra performance liquid chromatography(UPLC)analysis method that can simultaneously determine niacin,niacinamide,and vitamin B6 in formula milk powder.Gradient elution was performed with 10 mmol/L ammonium acetate solution(containing 0.1%formic acid)as mobile phase A and methanol as mobile phase B.The analytes were separated by Poroshell 120 EC-C_(18) in tandem with ACQUITY®UPLC HSS T3 column,detected with the UV detector and fluorescence detector,and quantified by an external standard method.Good linearity was observed for niacin concentrations in the range of 0.0-0.8μg/mL,nicotinamide concentrations in the range of 0.0-20.0μg/mL,pyridoxamine concentrations in the range of 0.0-0.2μg/mL,pyridoxal concentrations in the range of 0.0-0.6μg/mL and pyridoxine concentrations in the range of 0.0-4.0μg/mL,and the correlation coefficients of standard curves were all greater than 0.999.The limits of detection and quantification met the requirements.The average recoveries of the method were all in the range of 80.5%-104.1%with relative standard deviation(RSD)were 0.37%-2.99%.This method is simple,accurate,can be used for batch detection of niacin,niacinamide,and vitamin B6 in formula milk powder.