该研究建立了油料油脂中生育酚含量高效液相色谱法测定方法。样品加入异丙醇和0.1 g抗氧化剂2,6-二叔丁基对甲酚,经涡旋和超声提取,过0.22 μm的有机滤膜,采用Poroshell 120 SB-C18液相色谱柱分离,以90%(体积分数)甲醇-水为流动相、流速0.75 mL/min,柱温30 ℃,紫外检测波长294 nm,外标法定量分析。方法学考察结果显示,该方法10 min内可以实现4种生育酚基线分离。α-生育酚、β-生育酚、γ-生育酚、δ-生育酚的检出限分别是0.16、0.21、0.20、0.22 μg/g,定量限分别是0.53、0.68、0.66、0.72 μg/g,相关系数均大于0.999 2,且回收率为87%~109%,精密度为1.88%~9.89%。该方法提取过程简单且提取效率高,检测时间短,灵敏度高,重复性好,可满足实际油料样品中生育酚含量的检测需要。
In this study, analytical method of four tocopherols in oilseeds and edible oils was developed by reversed-phase high performance liquid chromatography. The sample was firstly dissolved by isopropanol and 0.1 g antioxidant (BHT) was added, then the mixture was extracted by vortex, ultrasonication and centrifugation. Finally, the mixture passed through 0.22 μm organic filter membrane for the purification. Poroshell 120 SB-C18 liquid chromatographic column was used with 90% methanol-water as mobile phase for the detection of tocopherols. The validation results indicated that the complete separation of four tocopherols could be achieved within 10 min by quantitative analysis on a high performance liquid chromatography with a flow rate of 0.75 mL/min, a column temperature of 30 ℃ and a detection wavelength of 294 nm. The recoveries of the four tocopherols were in the range of 87%-109%, and the precision (RSD) was in the range of 1.88%-9.89%. The limits of detection (LODs) for α-tocopherol, β-tocopherol, γ-tocopherol and δ-tocopherol were 0.16, 0.21, 0.20 and 0.22 μg/g, and the limits of quantification (LOQs) were 0.53, 0.68, 0.66 and 0.72 μg/g respectively. The correlation coefficients were higher than 0.999 2. This method has a simple extraction process, high extraction efficiency and sensitivity, which meets the actual sample detection requirements.
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