摘要
目的建立用半制备型反相高效液相色谱系统纯化、正相高效液相色谱硅胶柱色谱分离测定保健食品中维生素D_3含量的方法。方法通过皂化排除样品中的干扰物质,再经乙醚萃取浓缩,将浓缩后供试品溶液注入反相高效液相色谱仪净化分离去除杂质,排除维生素A的干扰,再用正相分析色谱进行定量分析,以正己烷-正戊醇(997:3,V:V)为流动相,流速为1.0 mL/min,采用色谱柱Waters Nova-Pak?Silia分离,在254 nm检测波长下经紫外或二极管阵列检测器检测,外标法定量。结果维生素D_3浓度在0.297~9.506μg/mL范围内呈良好的线性关系,相关系数为1,平均回收率为100.55%~100.58%,相对标准偏差值为0.44%~0.72%。结论该方法快速准确,可适用于保健食品中维生素D_3含量的测定。
Objective To establish a method for the determination of cholecalciferol in health food,which included purification with semi-preparative reversed liquid chromatography （RPLC） system,and separation with silica gel column of positive phase high performance liquid chromatography （HPLC）.Methods Samples were saponified to exclude the interfering substances,extracted with ether,and then concentrated.Subsequently,the concentrated test solution was injected into RPLC machine to separate and purify,so that the impurities could be removed,meanwhile,the interference brought by vitamin A would be eliminated.Finally,positive phase high performance liquid chromatography was employed for quantitative analysis,using hexane-pentanol （997∶3,V∶ V） as mobile phase at the flow rate of 1.0 mL/min,separated by Waters Nova-Pak（R）Silia column,and determined by ultraviolet detector or diode array detector at the wavelength of 254 nm with external standard method for quantification.Results Cholecalciferol had a good linear relationship in the range of 0.297～9.506 μg/mL with the correlation coefficient of 1.The average recovery and relative standard deviation （RSD） were 100.55%～100.58% and 0.44%～0.72%,respectively.Conclusion The method is rapid and accurate,which can be used for the determination of cholecalciferol in health food.
出处
《食品安全质量检测学报》
CAS
2017年第5期1622-1628,共7页
Journal of Food Safety and Quality
基金
湖南省自然科学基金项目(2016JJ6079)~~
关键词
维生素D3
高效液相色谱法
保健食品
cholecalciferol
high performance liquid chromatography
health food
