HPLC测定维生素ADE注射液中三种维生素的含量

Determination of 3 Kinds of Vitamins in Vitamin ADE Injection by HPLC

采用HPLC建立了兽药维生素ADE注射液中维生素A、维生素D3和维生素E的含量检测。采用C18色谱柱(150 mm×4.6 mm,5μm),甲醇为流动相A、乙腈-乙醇-水(60∶25∶5)为流动相B进行梯度洗脱,流速为1.5 m L/min,检测波长为265 nm,柱温为25℃,进样量为20μL。结果表明,在此色谱条件下,维生素ADE注射液中维生素A、维生素D3和维生素E的分离度良好,分别在60.96~1219.20μg/m L、2.56~51.20μg/m L和300.23~6004.60μg/m L范围内线性关系良好(r2>0.999)。按外标法以峰面积计算进行检测,维生素A、维生素D3和维生素E的平均回收率分别为99.6%、101.1%和99.0%,RSD均小于1.0%;平均重复性分别为105.7%、106.8%和102.3%,RSD均小于1.0%;中间精密度的RSD均小于0.5%;稳定性的RSD均小于1.0%;耐用性中选择流动相A与B变化为17∶83、流速为1.5 m L/min时,在一定范围内三种组分的含量基本稳定;柱温为25℃时,可避免维生素降解。说明该方法简单、快速,适用于维生素ADE注射液中三种维生素含量的测定,具有分离效果好、回收率高和稳定性好的特点。

To establish a HPLC method for determination of vitamin A,vitamin D3 and vitamin E in vitamin ADE injection. Using C18 column（150 mm×4.6 mm,5 μm） with a gradient elution of methanol as the mobile phase A and acetonitrile-ethanol-water（60 ∶ 25 ∶ 5） as the mobile phase B at a flow rate of 1.5 m L/min and a column temperature of 25 ℃. The determination wave length is 265 nm,while the injection volume is 20 μL. The results indicated that calibration curves for vitamin A,vitamin D3 and vitamin E were linear over the range of 60. 96 -1219.20 μg/m L,2. 56 - 51. 20 μg/m L and 300. 23 - 6004. 60 μg/m L（ r^2 〉0. 999）,respectively. The mean recoveries of vitamin A,vitamin D3 and vitamin E were 99.6%,101.1% and 99. 0%（ RSD 〈1. 0%）; the mean repetitive were 105.7%,106.8% and 102.3%（ RSD〈1.0%）; the RSD of intermediate precision were less than 0.5%; the RSD of stability were less than 1.0%. During the durability,when the vary of mobile phase（ 17 ∶ 83）and flow rate（1.5 m L/min） made the contents were basically stable,while column temperature is 25 ℃ to avoid the degradation of vitamin contents. This method has good selectively,high recovery and good stability,and can be used for the determination of vitamin ADE injection.