柱切换二维色谱法测定鱼肝油制剂中维生素D_(3)含量  

Determination of vitamin D_(3)content in cod liver oil using a column-switching technique

作　　者：戚绿叶 张丽媛 程巧鸳 颜琳琦 周明昊 QI Lyuye;ZHANG Liyuan;CHENG Qiaoyuan;YAN Linqi;ZHOU Minghao(Zhejiang Institute for Food and Drug Control,Hangzhou 310052,China)

机构地区：[1]浙江省食品药品检验研究院,浙江杭州310052

出　　处：《浙江大学学报(医学版)》2024年第6期779-784,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：国家药品标准提高项目。

摘　　要：目的:建立鱼肝油制剂中维生素D_(3)含量测定的柱切换二维液相色谱方法。方法:通过皂化萃取制备样品,采用双泵单阀切换双检测器的二维液相色谱法,对色谱柱、捕获装置和检测波长进行优化,对方法的线性、系统适用性、回收率、重复性和样品稳定性进行考察,并在实际样品测定中进一步验证。结果:建立了柱切换二维色谱法:一维色谱以Agilent PoroShell SB-C8(50 mm×4.6 mm,2.7μm)为色谱柱,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;二维色谱以Agilent Zorbax SB-C18(150 mm×3.0 mm,1.8μm)为色谱柱,乙腈为流动相,流速为0.42 mL/min,检测波长为264 nm。维生素D_(3)在线性范围内与峰面积成良好的线性关系,系统适用性、回收率、重复性、稳定性等均符合要求。结论:本研究建立的柱切换二维色谱法结果准确、重复性好、操作简单、检测效率高,并可同时基线分离前维生素D_(3)、反式维生素D_(3)、维生素D_(3)和速甾醇D_(3),适合推广应用。Objective:To develop a two-dimensional liquid chromatography method to determine the content of vitamin D_(3)in cod liver oil preparations.Methods:The samples were prepared by saponification and extraction,and the content of vitamin D_(3)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.Results:A column switching two-dimensional chromatography method was developed.In the first chromatography dimension,an Agilent PoroShell SB-C8(50 mm×4.6 mm,2.7μm)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,1.8μm)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_(3)content,there was a good linear relationship in the concentration range.The system suitability,recovery rate,repeatability and sample stability all met verification requirements.Conclusion:The two-dimensional liquid chromatography method developed in this study is accurate,reproducible and simple,and can simultaneously separate pre-vitamin D_(3),trans-vitamin D_(3),vitamin D_(3),and tachysterol D_(3).

关 键 词：鱼肝油 维生素D_(3) 含量测定 二维液相色谱法 

分 类 号：R284.1[医药卫生—中药学] R917.101[医药卫生—中医学]