建立RP-HPLC法同时测定复合维生素注射液中11种维生素含量

Simultaneous determination of 11 kinds of vitamins in compound vitamin injection by RP-HPLC

目的:建立同时测定复合维生素注射液中11种维生素含量的RP-HPLC方法。方法:采用十八烷基键合硅胶为填料(250 mm×4.6 mm,5μm)的色谱柱,以0.1 mol·L^(-1)的醋酸钠溶液-甲醇为流动相梯度洗脱,变化流速,联合紫外检测器的程序变化波长,进样量20μL,柱温35℃。结果:维生素A、维生素D_3、维生素E、维生素C、维生素B_1、维生素B_2、维生素B_6、维生素B_(12)、烟酸、烟酰胺、叶酸分别在0.272 8~2.728、0.012 4~0.124、1.032 7~10.327、1.131 0~11.310、2.282 8~22.828、0.194 8~1.948、1.016 5~10.165、0.235 8~2.358、1.085 4~10.854、2.168 1~21.681和0.369 4~3.694μg呈良好的线性关系(r>0.999);平均加样回收率(n=9)分别为100.0%、104.2%、100.9%、98.9%、100.4%、100.7%、100.1%、99.2%、100.2%、99.8%和100.5%,其RSD(n=9)分别为0.21%、0.02%、1.8%、1.0%、1.2%、0.35%、0.52%、2.8%、0.12%、0.05%和3.6%;精密度的RSD(n=6)分别为0.38%、0.11%、0.07%、0.15%、0.23%、0.06%、0.28%、0.22%、0.05%、0.41%、0.15%;重复性试验的RSD(n=6)分别为0.13%、0.78%、1.2%、2.2%、0.26%、1.5%、1.8%、1.3%、0.69%、0.94%和0.36%;样品平均含量分别为0.839、0.103、0.434、0.331、0.944、0.107、0.467、0.0043、0.134、0.958和0.013μg·μL^(-1)。结论:本方法适用于复合维生素注射液中维生素A、维生素D_3、维生素E、维生素C、维生素B_1、维生素B_2、维生素B_6、维生素B_(12)、烟酸、烟酰胺、叶酸的含量测定。

Objective： To established a method for simultaneous determination of eleven vitamins in compound vitamin injection byRP-HPLC.Method： The RP-HPLC was performed on C18 column（250 mm×4.6 mm,5 μm）.The mobile phase consisted of 0.1 mol·L-1 sodium acetate solution-methanol with the linear gradient elution,the changed flow rate,and changed wavelength of UV spectra,20μL was injected.The column temperature was 35℃.Results： vitamin A,vitamin D3,vitamin E, thiamine, vitaminB2, vitamin B6, vitamin B12, nicotinic acid, nicotinamide, folic acid had a good linearity among the ranges of 0.272 8-2.728,0.012 4-0.124,1.032 7-10.327,1.131 0-11.310,2.282 8-22.828,0.194 8-1.948,1.016 5-10.165,0.235 8-2.358,1.085 4-10.854, 2.168 1-21.681 and 0.369 4-3.694 μg,respectively; The average recoveries（n=9）were 100.0% with RSD of 0.21%,104.2% with RSD of 0.02%,100.9% with RSD of 1.8%,98.9% with RSD of 1.0%,100.4% with RSD of 1.2%,100.7% with RSD of 0.35%,100.1% with RSD of 0.52%,99.2% with RSD of 2.8%,100.2% with RSD of 0.12%,99.8% with RSD of 0.05%,100.5% with RSD of 3.6%,respectively. The RSDs of precisions were 0.38%, 0.11%,0.07%,0.15%,0.23%,0.06%,0.28%,0.22%,0.05%,0.41%,0.15%;the RSDs of repeatibility were 0.13%, 0.78%,1.2%,2.2%,0.26%,1.5%,1.8%,1.3%,0.69%,0.94%,0.36%,respectively;the average content was 0.839,0.103,0.434,0.331,0.944,0.107,0.467,0.004 3,0.134,0.958 and 0.013 μg·μL-1,respectively.Conclusion： This method can be used for simultaneous determination of vitamin A,vitamin D3,vitamin E,thiamine,vitamin B2,vitamin B6,vitamin B12,nicotinic acid, nicotinamide,folic acid in compound vitamin injection.