反相高效液相色谱法测定油类保健食品中混合生育酚

Determination of mixed tocopherols in oil health food by reversed-phase high performance liquid chromatography

目的建立测定油类保健食品中混合生育酚的反相高效液相色谱法。方法样品经氢氧化钾甲醇溶液皂化,盐酸甲醇溶液中和;用反相高效液相色谱:色谱柱(SUPELCO,C18,4.6 mm×150 mm,5μm);流速:1.0 m L/min;柱温:40℃;进样量:20μL;波长:294 nm;流动相:水和甲醇,梯度洗脱;将α-、γ-及δ-生育酚分离;外标法定量。结果本方法标准曲线在0.01~0.20 mg/m L范围内有良好的线性关系,r】0.9990,回收率为93.5%~103.8%,RSD【4.0%,检出限为0.02μg/kg,定量限为0.1μg/kg。结论该方法结果准确可靠,节省样品前处理时间及试剂,有利于降低检验成本,适用于油类保健食品中混合生育酚含量的测定。

Objective To establish a method for determination of mixed tocopherols in oil health foods by reversed-phase high performance liquid chromatography. Methods The samples were saponified by potassium hydroxide methanol, and neutralized by hydrochloric acid methanol. The extracts were separated by RP-HPLC with the column of SUPELCO, C18 (4.6 mm×150 mm, 5 μm), flow rate of 1.0 mL/min, column temperature of 40℃, injection volume of 20μL, wavelength of 294 nm, mobile phase of water and methanol, and with gradient elution to separateα-,γ-andδ-tocopherol;then were determined by external standard method. Results Tocopherols standard curve had a good linear relationship within 0.01~0.20 mg/mL range (r>0.9990), the recovery was 93.5%~103.8%, RSD<4.0%, limit of determination was 0.02μg/kg, and the limit of quantity was 0.1μg/kg. Conclusion The method is accurate and reliable, saving pretreatment time and reagents, can reduce the test cost;it is suitable for the determination of mixed tocopherol in oil health food.