固相萃取-反相高效液相色谱法同时测定饲料中的角黄素和虾青素

建立了一种同时测定饲料中角黄素和虾青素的固相萃取-高效液相色谱法（HPLC）。样品由乙腈提取,经LC-NH2固相萃取小柱净 化,洗脱剂为乙腈-甲苯（体积比为3∶1）,洗脱液被浓缩后进行HPLC分析,色谱柱为ZORBAX Eclipse XDB-C18柱(150 mm×4.6 mm,5 μ m),流动相为乙腈-甲醇（体积比为95∶5）,流速1.0 mL/min,采用二极管阵列检测器检测,检测波长为474 nm;外标法定量。角黄素和 虾青素的线性范围分别为1.0～30.0 mg/L和1.0～20.0 mg/L,相关系数分别为0.9990和0.9991,回收率为90%～101%,相对标准偏差为 0.62%～3.68%,检出限分别为0.84和0.60 mg/L。该方法简便、快速、准确,可用于饲料中角黄素和虾青素的同时测定。

Simultaneous determination of canthaxanthin and astaxanthin in feedstuffs using solid phase extraction-reversed- phase high performance liquid chromatography

A method was developed for the simultaneous determination of canthaxanthin and astaxanthin in feedstuffs using reversed-phase high performance liquid chromatography (RP-HPLC). The sample was extracted by acetonitrile, and cleaned up by an LC-NH2 column. An Agilent ZORBAX Eclipse XDB-C18 analytical column (150 mm x 4.6 mm, 5 microm) was used and kept at 25 degrees C. Acetonitrile-methanol (95 : 5, v/v) was used as the mobile phase at a flow rate of 1.0 mL/min. The detection was performed by a diode array detector at 474 nm. The quantitive analysis of external standard calibration curves was used. The linear ranges of the method for canthaxanthin and astaxanthin were 1.0 - 30.0 mg/L (r = 0.999 0) and 1.0 - 20.0 mg/L (r = 0.999 1), respectively. The average recoveries were 90% - 101% with the relative standard deviations of 0.62% - 3.68%. The detection limits were 0.84 and 0.60 mg/L for canthaxanthin and astaxanthin, respectively. The method is simple, precise, sensitive and reproductive. It can be used to determine the contents of canthaxanthin and astaxanthin in feedstuffs.