高效液相色谱法同时测定血清中VA、VD_3、VE和β-胡萝卜素

Detecting VA,VD_3,VE and β-carotene with HPLC in Human Serums

目的建立能同时测定人血清中维生素A(VA)、维生素D3(VD3)、维生素E(VE)和β-胡萝卜素的高效液相色谱(high performance liquid chromatography,HPLC)法。方法以无水乙醇沉淀蛋白质后,血清中的脂溶性维生素用乙醚/石油醚混合溶液提取,吹氮浓缩后用HPLC测定。结果VA、VD3、VE和β-胡萝卜素的线性范围分别为0.012μg/mL^500μg/mL、0.030μg/mL^500μg/mL、0.12μg/mL^500μg/mL和0.015μg/mL^500μg/mL;检出限分别为0.012μg/mL、0.030μg/mL、0.12μg/mL和0.015μg/mL;相对标准偏差分别为0.75%、0.54%、2.06%和2.74%。血清样品中VA、VD3、VE和β-胡萝卜素的回收率分别为:92%~116%,98%~112%,84.8%~106%和90%~105%。结论本法简便快速,适于血清样品中4种脂溶性维生素的同时测定。

Objective To develop a method of detecting retinol（vitamin A）,vitamin D3,α-tocopherol（vitamin E） and β-carotene in human serums with HPLC.Methods Proteins were precipitated with anhydrous ethanol.Fat-solutable vitamins in human serums were extracted with aether/petroleum ether mixture and determined with HPLC.Results The linear ranges for the retinol,VD3,α-tocopherol and β-carotene were 0.012 μg/mL-500 μg/mL,0.030 μg/mL-500 μg/mL,0.12 μg/mL-500 μg/mL,and 0.015 μg/mL-500 μg/mL,respectively.The detection limits for the retinol,VD3,α-tocopherol and β-carotene were 0.012 μg/mL,0.030 μg/mL,0.12 μg/mL and 0.015 μg/mL,respectively.The relative standard derivations（RSD） for the retinol,VD3,α-tocopherol and β-carotene were 0.75%、0.54%、2.06% and 2.74%,respectively.The proposed method recovered 92%-116%,98%-112%,84.8%-106%,and 90%-105% retinol,VD3,α-tocopherol and β-carotene in human serums respectively.Conclusion The method is simple,quick and applicable to all of the four fat-solutable vitamins.