Qi Lyuye, Zhang Liyuan, Cheng Qiaoyuan, Yan Linqi, Zhou Minghao
Zhejiang Institute for Food and Drug Control, Hangzhou 310052, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2024 Dec 25;53(6):779-784. doi: 10.3724/zdxbyxb-2024-0045.
To develop a two-dimensional liquid chromatography method to determine the content of vitamin D in cod liver oil preparations.
The samples were prepared by saponification and extraction, and the content of vitamin D 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 chromatography dimension, an Agilent PoroShell SB-C8 (50 mm×4.6 mm, 2.7 μm) column was used with acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1.0 mL/min and detection wavelength as 264 nm. An Agilent Zorbax SB-C18 (150 mm×3.0 mm,1.8 μm) column was used in the second chromatography dimension with acetonitrile as the mobile phase at a flow rate of 0.42 mL/min, and detection wavelength as 264 nm. In determining vitamin D content, there was a good linear relationship in the concentration range. The system suitability, recovery rate, repeatability and sample stability all met verification requirements.
The two-dimensional liquid chromatography method developed in this study is accurate, reproducible and simple, and can simultaneously separate pre-vitamin D, trans-vitamin D, vitamin D, and tachysterol D.
建立一种二维液相色谱法测定鱼肝油制剂中维生素D的含量。
样品经皂化和萃取处理，采用双泵单阀切换双检测器二维液相色谱法测定维生素D含量。对色谱柱、捕集装置和检测波长进行优化；考察该方法的线性、系统适用性、回收率、重复性和样品稳定性，并在实际样品测定中进一步验证。
建立了柱切换二维色谱法。在第一维色谱中，采用安捷伦PoroShell SB-C8（50 mm×4.6 mm，2.7μm）色谱柱，以乙腈-水为流动相，梯度洗脱，流速为1.0 mL/min，检测波长为264 nm。在第二维色谱中，采用安捷伦Zorbax SB-C18（150 mm×3.0 mm，1.8μm）色谱柱，以乙腈为流动相，流速为0.42 mL/min，检测波长为264 nm。在测定维生素D含量时，浓度范围内线性关系良好。系统适用性、回收率、重复性和样品稳定性均符合验证要求。
本研究建立的二维液相色谱法准确、重现性好且操作简便，能同时分离维生素原D、反式维生素D、维生素D和速甾醇D。
