反相高效液相色谱法测定果醋中的8 种维生素

建立梯度洗脱反相高效液相色谱法同时检测果醋中VC、VB1、VB2、VB3、VB6、VB12、叶酸和烟酰胺8种水溶性维生素的分析方法。考察柱温、缓冲液浓度、pH值以及梯度洗脱程序对各组分分离的影响。最终采用0.1mol/L醋酸钠溶液(pH4.25)和甲醇作为流动相进行梯度洗脱，流速为1.00mL/min，柱温为30℃，8种水溶性维生素在14.2min内达到基线分离；紫外检测波长为266nm和280nm；线性范围：VB2、叶酸为0.05～12.5μg/mL；VC、VB1、VB3、烟酰胺为0.1～25μg/mL；VB6、VB12为0.15～37.5μg/mL，相关系数均在0.999以上，其最低检出限为0.47～3.46ng/mL。该法被成功地用于定量果醋中的维生素，平均加标回收率为92.4%～105.2%，相对标准偏差为0.08%～1.54%。

Determination of Eight Water-Soluble Vitamins in Fruit Vinegar by RP-HPLC

A RP-HPLC gradient elution method was developed for the simultaneous determination of eight water-soluble vitamins,including ascorbic acid,thiamine,riboflavin,nicotinic acid,pyridoxine,cyanocobalamin,folic acid and nicotinamide in fruit vinegar.The effects of temperature,buffer solution concentration,pH value and gradient elution procedure on chromatographic separation were studied.The mobile phase was a mixture of 0.1 mol/L sodium acetate solution(pH 4.25) and methanol at a flow rate of 1.00 mL/min.The column temperature was set as 30 ℃.Baseline separation was achieved for 8 water-soluble vitamins within 14.2 min.The detection was carried out at 266 nm and 280 nm.The method offered excellent linearity over a range of 0.05-12.5 μg/mL for riboflavin and folic acid,0.1-25 μg/mL for ascorbic acid,thiamine,nicotinic acid and nicotinamide,0.15-37.5 μg/mL for pyridoxine and cyanocobalamin,with correlation coefficients(r>0.999).The detection limits were in the range of 0.47-3.46 ng/mL This method has been successfully applied to assay vitamins in fruit vinegar.The average spike recovery varied from 92.4% to 105.2% with RSD in the range of 0.08%-1.54%(n =3).