反相高效液相色谱法同时测定鸡血清中视黄醇和α—生育酚

Simultaneous Determination of Retinol and α- Tocopherol in Chicken Serum by RP- HPLC

建立反相高效液相色谱法同时测定鸡血清中视黄醇和α-生育酚的方法。取血清100μL，加入脱醛无水乙醇100μL，振荡，加入正己烷萃取，将正己烷吹干后用甲醇溶解，进样。色谱条件为：色谱柱Agilent ZORBAXSB—C18柱（150mm×4．6mm，5μm）；流动相为甲醇：水（98：2）；检测波长视黄醇325nm，α-生育酚285nm。结果显示视黄醇在0．025—2．0μg／mL浓度范围内呈线性，r^2=0．99999，α-生育酚在0．5—20．0μg／mL浓度范围内呈线性关系，r^2=0．9996；视黄醇和α-生育酚的平均回收率分别为94．5％和93．3％。因此，使用本方法选择不同波长同时测定鸡血清中视黄醇及α-生育酚的含量，方法快速、灵敏、准确，样品处理简便易行。

To establish a method to determinate simultaneously the concentrations of retinol and α -tocopherol in chiken serum. 100μL serum was added 100μL anhydrous alcohol and 0.7 mL hexane, the mixture were shaken and centrifuged, then dried hexane with N2 gas. Resolved with methanol. The HPLC system was consisted of Agilent ZORBAX SB - C18 （ 150mm × 4.6mm, 5μm）. Mobile phase was methanol： water （98：2）, detected wavelenth was 325nm and 285nm. The results showed that the linear ranges of determination of refinol and α - tocopherol were 0.025 - 2.0μg/mL （ r^2 = 0. 99999） and 0. 5 - 20.0μg/mL（ r^2 = 0. 9996） respectively. The average recovery rate were 94.5% for retinol and 93.3% for α - tocopherol. Therefore ,the method is sensitive, simply, rapid and accurate.