二维色谱法测定维生素AD软胶囊和滴剂中维生素D的含量

Determination of vitamin D in vitamin AD soft capsules and drops by two-dimensional liquid chromatography

目的:建立维生素AD软胶囊和滴剂中维生素D含量测定的二维液相色谱的检测方法。方法:采用三氯甲烷和异丙醇直接溶解样品,以双三元泵双阀切换单检测器的二维液相色谱法测定维生素D含量。一维色谱以AcclaimTMC8(150 mm×3.0 mm,3μm)为色谱柱,乙腈-甲醇-水为流动相梯度洗脱,流速为0.8 mL·min^-1,检测波长为264 nm;二维色谱以AccucoreTMpolar premium (150 mm×4.6 mm,2.6μm)为色谱柱,乙腈-甲醇-水为流动相梯度洗脱,流速为0.5 mL·min^-1,检测波长为264 nm。结果:维生素D2,D3分别在0.248~3.968,0.258~4.128μg·mL^-1浓度范围内与峰面积具有良好的线性关系(r=1.000 0,0.999 7),回收率在99.54%~101.97%,99.93%~101.59%之间。3批维生素AD软胶囊测得维生素D2含量分别为107.92%,110.37%和109.90%,3批维生素AD滴剂测得维生素D3含量分别为96.74%,97.53%和97.77%。结论:新建的二维色谱法结果准确、重复性好、操作简单,可用于维生素AD软胶囊和滴剂中维生素D2/D3含量的直接测定。

Objective: To establish a two-dimensional liquid chromatography method to determine the content of vitamin D in vitamin AD soft capsules and drops.Methods: The sample was dissolved in trichloromethane and diluted with 2-propanol,and the content of vitamin D was determined by two-dimensional liquid chromatography with dual ternary pump dual valve switching single detector.In the first dimensional chromatography,an AcclaimTMC8 (150 mm × 3.0 mm,3 μm) column was used,and acetonitrile-methanol-water was used as mobile phase and gradiently eluted at a flow rate of 0.5 mL·min^-1.The detection wavelength was 264 nm.An AccucoreTMpolar premium (150 mm × 4.6 mm,2.6 μm) column was used in the second dimensional chromatography,and acetonitrilemethanol-water was used as mobile phase and gradiently eluted at a flow rate of 0.8 mL·min^-1.The detection wavelength was 264 nm.Results: The calibration curves of VD2 and VD3 were linear in the concentration range of0.248 ~ 3.968 and 0.258 ~ 4.128 μg·mL^-1 (r = 1.000 0 and 0.999 7),respectively.The recoveries were99.54% ~ 101.97% and 99.93% ~ 101.59%,respectively.The contents of vitamin D in three batches of vitamin AD soft capsules and three batches of vitamin AD drops were measured to be 107.92%,110.37%,109.90% and 96.74%,97.53%,and 97.77%,respectively.Conclusion: The established two-dimensional liquid chromatography method is accurate,simple and rapid,and can be used to determine the content of vitamin D directly in vitamin AD soft capsules and drops.