反相高效液相色谱-二极管阵列检测蛋黄中的角黄素和虾青素

目的建立蛋黄中角黄素及虾青素的反相高效液相色谱-二极管阵列同时检测法,探讨样品前处理方法及色谱条件的优化。方法蛋黄样品以乙腈超声提取,冷冻除脂,固相萃取净化并氮吹定容后以C18柱分离,乙腈为流动相,二极管阵列检测器于470nm波长下测定。结果实验表明角黄素及虾青素在0.5～100μg/mL浓度范围内线性关系良好(r≥0.998),日内精密度≤3.6%,日间精密度≤5.2%,两者平均加标回收率分别为91.5%及88.7%,检出限分别为0.035μg/mL及0.027μg/mL。结论本法简便快速,灵敏度高,易于推广,适用于蛋黄中角黄素及虾青素的同时检测。

Determination of canthaxanthin and astaxanthin in egg yolks by reversed phase high performance liquid chromatography with diode array detection

Objective A method using reversed phase high performance liquid chromatography(RP-HPLC) coupled with diode array detector(DAD) was developed for the simultaneous determination of canthaxanthin and astaxanthin in egg yolks.Methods Samples were extracted with acetonitrile in ultrasonic bath for 20 minutes and then purified by freezing-lipid filtration and solid phase extraction(SPE).After being vaporized to dryness by nitrogen blowing and made up to volume with methanol,the extract solution was chromatographically separated in C18 column with a unitary mobile phase consisting of acetonitrile.The proposed method was validated in terms of linearity,precision,accuracy,and limit of detection(LOD).Result Regression analysis revealed a good linearity between peak area of each analyte and its concentration(r≥0.998).The intra-and inter-day relative standard deviations(RSDs) were less than 3.6% and 5.2%,respectively.LODs of canthaxanthin and astaxanthin were 0.035 and 0.027 μg/mL(S/N=3).The average recoveries of canthaxanthin and astaxanthin were 91.5% and 88.7%.Conclusion The proposed method is simple,fast and easy to apply.