Determination of vitamin D3 content in cod liver oil using column-switching technique.

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To develop a two-dimensional liquid chromatography method to determine the content of vitamin D3 in cod liver oil preparations. The samples were prepared by saponification and extraction, and the content of vitamin D3 was determined by two-dimensional liquid chromatography with dual-pump-single-valve switching dual detectors. The chromatographic column, capture device, and detection wavelength were optimized; the linearity, system suitability, recovery rate, repeatability and sample stability of the method were investigated, and further validated in actual sample determination. A column switching two-dimensional chromatography method was developed. In the first dimensional chromatography, an Agilent PoroShell SB-C8 (50 mm×4.6 mm, 2.7 μm) column was used, and acetonitrile-water was used as mobile phase and gradiently eluted at a flow rate of 1.0 mL/min, the detection wavelength was 264 nm. An Agilent Zorbax SB-C18 (150 mm×3.0 mm,1.8 μm) column was used in the second dimensional chromatography, and acetonitrile was used as mobile phase at a flow rate of 0.42 mL/min, the detection wavelength was 264 nm. In determining content of Vitamin D3, there was a good linear relationship in the concentration range and the system suitability, recovery rate, repeatability and sample stability all met the verification requirements. The developed two-dimensional liquid chromatography method in the study is accurate, reproducible and simple, it can simultaneously separate pre-vitamin D3, trans-vitamin D3, vitamin D3, and tachysterol3, the baseline separation was achieved, indicating that it is worth promoting for application.