柱前衍生-高效液相色谱法测定食物和保健食品中维生素B_1

Determination of Vitamin B_1in Food and Health Food by High Performance Liquid Chromatography Coupled with Pre-column Derivatization

目的：建立柱前衍生-高效液相色谱法（HPLC）测定食品中维生素B-含量的方法。方法：样品以0．1mol／L盐酸在120℃条件下高压提取，经2％碱性铁氰化钾溶液衍生后，以正丁醇液液萃取，用HPLC测定萃取物。采用SymmetryC18柱（150mm～4．6mm，5μm）分离，以0．05mol／L乙酸钠溶液-甲醇（体积比65：35）为流动相，荧光检测器检测（Ex=375nm，Em=435nm），外标法定量。结果：维生素B1在0．1～1．0mg／L范围内线性关系良好，相关系数大于0．9990；不同食品中维生素B1的检出限为0．2—0．3mg／kg，定量限为0．6—0．9mg／kg；对鸡蛋等4种食品基质进行加标试验，维生素B1的回收率均值为90．1％～109％，相对标准偏差（RSD）为1．9％-8．5％（n=6）。结论：该方法简单快速，灵敏度高，准确性好，可适用于各类食品和保健食品中维生素B1含量的测定。

Objective：A simple method based on pre-column derivatization coupled with high performance liquid chromatography （HPLC） was developed for the determination of vitamin B1 in foods and health foods. Methods：The ho- mogenized food sample was extracted with 0.1 mol/L hydrochloric acid at 120℃ and the extract or the cleaned up solu- tion was derivatized with 2% alkaline potassium ferricyanide solution. The separation of the vitamin B1 deravatives extracted by butanol was achieved on a Symmetry C18 column （150 mm×4.6 mm,5μm）with 0.05 mol/L sodium acetate- methanol （65：35,V/V） as the mobile phase. The analyte was detected by the fluorescence detector （Ex--375 nm, Em=435 nm）. The quantification was achieved by the external standard method. Results：Good linearity was observed in the range of 0.1-1.0 mg/L with the correlation coefficient 〉0.9990. The limits of detection and quantification for vitamin B1 were in the range of 0.2-0.3 mg/kg and 0.6-0.9 mg/kg for the selected foods, respectively. The average recoveries of vitamin B1 spiked in fresh egg, apple, potato and amcal were 90.1%-109% and the relative standard deviation （RSD） was 1.9%-8. 5%（n=6）. Conclusion：Owing to the simplicity, rapidity, high sensitivity and accuracy, the method can be widely used for the quantification of vitamin B1 in the various foods and health foods.