高效液相色谱法测定红小丑鱼中虾青素和叶黄素含量

Determination of lutein and astaxanthin in Amphiprion frenatus by reversed-phase high performance liquid chromatography

为建立检测红小丑鱼皮肤和鳍组织中虾青素、叶黄素含量的高效液相色谱分析方法。样品经丙酮提取,甲基叔丁基醚萃取,纯水分离杂质,LC-NH2净化,C18色谱柱分离,流动相为甲醇∶水(98∶2,v/v),紫外检测波长475 nm,外标法定量。虾青素和叶黄素标准品0.02~1μg/m L线性关系良好,虾青素相关系数为0.998,叶黄素相关系数为0.999,加标回收率分别为79.92%~90.32%和78.51%~97.60%,相对标准偏差分别为6.35%~7.07%和10.79%~13.41%。该方法前处理简单,能同时测定红小丑鱼组织中虾青素和叶黄素含量。

A high performance liquid chromatography method was established for the determination of lutein and astaxanthin in Amphiprion frenatus.The sample was extracted by acetone and methyl tertiary butyl ether,and cleaned up by an LC-NH2 column.The chromatographic separation was achieved on C18 column used amobile phase made up of methanol and water（98∶ 2,v/v） by equal gradient elution.The detection wave length was set at 475 nm,and an external standard method was used to quantify astaxanthin and lutein.An excellent linear relationship of this method for astaxanthin and lutein were observed with values of 0.998 and 0.999. The average spike recovery rates of lutein and astaxanthin were 79.92% ~ 90.32% and 78.51% ~97.60% with relative standard deviations of 6.35% ~ 7.07% and 10.79% ~ 13.41%. This method was sensitive,repeatable and suitable for the determination of astaxanthin and lutein in small ornamental fish.