高效液相色谱法测定血清叶黄素和玉米黄素

目的:建立C30-高效液相色谱(high performance liquid chromatography,HPLC)内标法测定血清叶黄素、玉米黄素和β-胡萝卜素。方法:应用Develosil C30-UG色谱柱(250 mm×4.6 mm,5μm),流动相A为乙腈∶甲醇(体积比60∶40),流动相B为甲基叔丁基醚,梯度洗脱。流速1 mL/min,检测波长450 nm,进样量20μL,柱温25℃。内标物为全反式-β-阿扑-8'-胡萝卜醛。结果:血清叶黄素、玉米黄素和β-胡萝卜素的分离效果良好,保留时间分别为9.9min、10.3 min和21.2 min,日内相对标准差分别为3.22%、3.81%和1.60%。血清叶黄素、β-胡萝卜素在0.012 5～12.5 mg/L(r=0.999 5、r=0.999 7),玉米黄素在0.005～5.0 mg/L(r=1)范围内呈良好线性关系。58名正常老年人(年龄50岁)血清叶黄素、玉米黄素和β-胡萝卜素的平均浓度分别为0.410μmol/L、0.054μmol/L和0.128μmol/L。结论:该C30-HPLC内标法是定量分析血清叶黄素、玉米黄素和β-胡萝卜素的可靠方法之一。

Determination of lutein and zeaxanthin in serum by high performance liquid chromatography

Objective:To establish a simultaneous determination method for measuring lutein,zeaxanthin and β-carotene in serum by internal standard on C30-HPLC.Methods: Experimental data were as follows: stationary phase,Develosil carotenoid column C30(250 mm×4.6 mm,5 μm);mobile phase A,acetonitrile ∶ methanol(3∶ 2,v/v);mobile phase B,MTBE;grads elution;flow rate,1 mL/min;monitoring wavelength,450 nm;injection volume,20 μL;column temperature,25 ℃.Results: Lutein,zeaxanthin and β-carotene were thoroughly separated with the average retention time of 9.9 min,10.3 min and 21.2 min,respectively.The intra-day relative standard deviation(RSD) values were 3.22%,3.81% and 1.60%.The linear ranges of serum concentrations of lutein and β-carotene were both 0.012 5-12.5 mg/L(r=0.999 5,r=0.999 7),and that of zeaxanthin was 0.005-5.0 mg/L(r=1).The mean serum concentrations of lutein,zeaxanthin and β-carotene for 58 healthy elder inhabitants(>50 years) were 0.410 μmol/L,0.054 μmol/L and 0.128 μmol/L,respectively.Conclusion: This established method can be used for determination of lutein,zeaxanthin and β-carotene in serum.