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

关键词:  HPLC  维生素ADE注射液  含量测定

Key words:  HPLC  vitamin ADE injection  content determination

摘要:

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，5 μm），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 mL/min，检测波长为265 nm，柱温为25 ℃，进样量为20 μL。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60.96～1219.20 μg/mL、2.56～51.20 μg/mL和300.23～6004.60 μg/mL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 mL/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。

Abstract:

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 mL/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/mL, 2.56~51.20 μg/mL and 300.23~6004.60 μg/mL (r2>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 mL/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.