高效液相色谱-荧光检测法测定保健食品中维生素K2的含量

Determination of the vitamin K2 content in functional food by high performance liquid chromatography-fluorescence detector method

中文关键词:  高效液相色谱-荧光检测法  保健食品  维生素K2  四烯甲萘醌

英文关键词:high performance liquid chromatography-fluorescence detector method  functional food  vitamin K2  menatetrenone 4

中文摘要:

      目的 建立高效液相色谱-荧光检测法快速检测保健食品中维生素K2(四烯甲萘醌)的分析方法。方法 样品使用GIST C18色谱柱(250 mm×4.6 mm, 5 μm)分离, 柱后经锌粉还原柱(4.6 mm×50 mm, 50～70 μm)衍生, 以甲醇(含有四氢呋喃10%、冰醋酸0.03%、氯化锌1.5 g/L、无水乙酸钠0.5 g/L)为流动相, 流速为1.0 mL/min, 进入荧光检测器进行检测。结果 四烯甲萘醌保留时间约为10 min, 在浓度0.1099～2.197 μg/mL范围内线性关系良好, 相关系数r2=0.9999。重复性实验RSD为0.794%, 3个浓度级别的加标回收率在97.1%～101.9%之间。当取样量为1.5 g, 总定容体积为500 mL时, 四烯甲萘醌的检出限为71 μg/100 g, 定量限为236 μg/100 g。结论 该方法准确性高、重现性好、操作简单, 可用于保健食品中维生素K2的含量测定。

英文摘要:

      Objective To establish an analytical method for the rapid detection of vitamin K2 (menaquinone 4) in functional food by high performance liquid chromatography-fluorescence detector. Methods The samples were separated by a GIST C18 column (250 mm×4.6 mm, 5 μm) and derivatized with a post-column zinc powder reduction column (4.6 mm × 50 mm, 50-70 μm). Methanol (containing 10% tetrahydrofuran, 0.03% glacial acetic acid, 1.5 g/L zinc chloride and 0.5 g/L anhydrous sodium acetate) was used as mobile phase at the flow rate 1.0 mL/min for detection by fluorescence detector. Results The retention time of menaquinone 4 was around 10 min. It had a good linearity in the range of 0.1099 to 2.197 μg/mL with the correlation coefficient (r2=0.9999). The RSD of the repeatability experiment was 0.794%. The recoveries for the 3 levels of concentration were between 97.1% and 101.9%. When the sample size was 1.5 g and the total volume was 500 mL, the detection limit of menaquinone 4 was 71 μg/100 g and the quantitative limit was 236 μg/100 g. Conclusion This method is accurate, reproducible and easy to operate. It can be used for the determination of vitamin K2 in functional food.