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Discovery C18色谱柱(150 mm×4.6 mm i.d.,5 um),以乙腈-甲醇为流动相进行梯度洗脱,检测波长为450 nm.结果:叶黄素和β-胡萝卜素在0.1～10.0 mg/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25 min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b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 evaporated 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 p-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 separation was completed within 25 min. CONCLUSION: This established method is accurate and reproducible. It can be used for the analysis of lutein and β-carotene in serum.
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