健康成人血清中叶黄素和β-胡萝卜素的HPLC法测定

目的：采用高效液相色谱（HPLC）建立同时测定健康成人血清中叶黄素和β-胡萝卜素的方法．方法：血清经蛋白沉淀，正己烷萃取，氮气吹干及流动相溶解等预处理过程后，进行高效液相色谱分析．采用DiscoveryC18色谱柱（150mm×4．6mm．d．，5μm），以乙腈-甲醇为流动相进行梯度洗脱，检测波长为450nm．结果：叶黄素和β-胡萝卜素在0．1～10．0mg／L范围内与峰面积呈良好线性关系，相关系数r分别为0．9996，0．9985（P〈0．05）．叶黄素和β-胡萝卜素的平均加样回收率在93．0％-103．9％之间，相对标准偏差（RSD）分别为1．19％和2．70％，分析周期在25min以内．结论：该方法灵敏度高，重现性好，是定量测定血清中叶黄素和β-胡萝卜素定量测定的可靠方法之一．

Determination of serum lutein and B-carotene by high performance liquid chromatography in healthy Chinese subjects

AIM： To develop a method for the simultaneous determination of serum lutein and β-carotene levels in healthy Chinese subjects by high performance liquid chromatography （HPLC）. METHODS： Lutein and β-carotene were extracted by hexane after protein precipitation. The organic layer was evaporat- ed under N2 and the residue was dissolved in mobile phase. Serum lutein and β-carotene levels were determined by HPLC. Discovery C18 column （ 150 mm × 4.6 mm i. d. , 5 μm） was employed. The mobile phase consisted of acetonitrile and metha- nol with the gradient elution. The detection wavelength was set at 450 nm. RESULTS： Lutein and β-carotene showed good linear relationships in the range of 0.1-10.0 mg/L, r = 0.9996 and r = 0.9985 （ P 〈 0.05 ）. The recoveries of lutein and β-carotene were 93.0%-103.9% , and RSDs were 1.19%-2.70%. The separa- tion was completed within 25 min. CONCLUSION： This estab- lished method is accurate and reproducible. It can he used for the analysis of lutein and β-carotene in serum.