高效液相色谱法同时测定血清中几种脂溶性维生素

THE SIMULTANEOUS DETERMINATION OF SEVERAL FAT-SOLUBLE VITAMINS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

目的:采用高效液相色谱(HPLC)技术同时测定维生素A(VA)、维生素E(VE)、β-胡萝卜素(β-C)和番茄红素(Ly)。方法:HPLC柱:AlltechODS5μm(250mm×4.6mmi.d.);流动相:甲醇∶乙酸乙酯∶正己烷=7∶2∶2(v∶v∶v);检测波长:VA,VE,β-C和Ly的检测波长分别为325nm、292nm、450nm和472nm(波长编程,自动转换)。灵敏度:0.002AUFS;流速:1.0ml/min。结果:VA,VE,Ly和β-C的保留时间分别是4.5,6.5,9.4和13.4min,4种成分峰独立完整,无干扰峰,峰形和分离效果良好,并在15min之内可完成分析测定。结论:本研究首次建立了一次同时测定VA、VE、β-C和Ly的方法,既节省时间和试剂,又快速、准确。

Objective： A new method of simultaneous determination of retinol （VA）, α-tocopherol （VE）, β-carotene （β-C） and lycopene （Ly） by high-performance liquid chromatography （HPLC） was developed. Method： The HPLC system consisted of chromatography column, analytical columm Alltech ODS 5μm （250mm×4.6mm i.d.）, and eluent, methanol ： ethylacetate ： n -hexane=7 ： 2 ： 2 （v ： v ： v）. The flow rate was 1.0 ml/min. The detector was programmed as follows, 4.5 min, 6.5 min, 9.4 min and 13.4 min, from 0 to 5 min at 325 nm to determine VA, from 5 to 7 min at 292 nm to determine VE, from 7 to 10 min at 472 nm to determine lycopene and from 10 to 15 min at 450 nm to determine β-carotene. Result： The retention time of VA, VE, β-C and Ly were 4.5, 6.5, 9.4 and 13.4 min, respectively. Four compounds were separated properly and the peaks were perfect. Conclusion： The method of simultaneous determination of VA, VE, β-C and Ly is established. This method is rapid and precise, and can be used in clinical test and experimental research.