高效液相色谱法测定血清叶黄素、番茄红素和β-胡萝卜素

DETERMINATION OF LUTEIN,LYCOPENE,AND β-CAROTENE IN SERUM BY HPLC

目的建立血清中叶黄素、β-胡萝卜素、番茄红素的高效液相色谱测定法。方法Inertsil ODS C18色谱柱（250×4.6mm,粒径5μm）;选用甲醇-乙腈-四氢呋喃（60：20：20）,含0.01%的抗坏血酸为流动相;流速为1.0ml/min;检测波长为450nm。结果叶黄素、番茄红素和β-胡萝卜素的保留时间分别为4.3min、12.4min和17.0min,回收率分别为97.98%～102.80%、97.24%～101.76%、98.56%-100.4%,日内和日间RSD分别为2.26%和3.02%、4.34%和5.02%、1.81%和2.52%。标准曲线的线性范围为0.05～2.00μg/ml。90名正常人血清中叶黄素、番茄红素与β-胡萝卜素的含量分别为0.34±0.17μg/ml、0.28±0.16μg/ml、0.49±0.25μg/ml。结论HPLC法测定人血清叶黄素、番茄红素和β-胡萝卜素浓度方法可靠,样品处理快速简便。

Objective To establish the HPLC method for accurate and precise measurement of the major carotenoids in plasma using HPLC detection. Method The chromatographic analysis was performed using an analytical scale C18 reversed phase column （250 mm×4.6 mm i.d.） The mobile phase for chromatography was methanol：acetonitrile： tetrahydrofuran （60：20：20） containing 0.01% ascorbic acid. The flow rate was set at 1 ml/min. Chromatographic runs were monitored at 450nm. Results The average retention time for lutein, lycopene and β-carotene were 4.3 min, 12.4 min and 17.0 min respsetivuly. The recovery rate was as follows：lutein 97.98%- 102.80%, lycopene 97.24%- 101.76%, β-carotene 98.56%- 100.4%. The intra-day and inter-day RSD were 2.26% and 3.02%, 4.34% and 5.02%, 1.81% and 2.52% for lutein, lycopene, and β-carotene respectively. Ninety healthy inhabitants were enrolled in this study. Mean plasma concentrations of lutein, lycopene and β-carotene were 0.34±0.17 μg/ml, 0.28±0.16 μg/ml, 0.49±0.25μg/ml. Conclusion This method was simple, rapid and accurate for determination of lutein, lycopene and β-carotene in serum.