β-葫萝卜素血浓度测定方法的研究

STUDIES ON THE METHODS OF BLOOD β-CAROTENE CONCENTRATION DETERMINATION

建立了用高效液相色谱法(HPLC)及可见紫外分光光度(VIS-UV)单波长法、双波长法,测定血浆中β-胡萝卜素(β-C)的浓度。在氯仿或乙醇溶液中检测限为5ng,人血浆中最低检测浓度为25ng/ml,平均回收率为98.8％,CV ％为3.2％。HPLC法的线性范围为25～2000 ng/ml血浆,UV单波长法线性范围为50～4000 ng/ml血浆,双波长法线性范围为100～4000ng/ml血浆。HPLC法与UV法的相关系数>0.99,斜率接近1。

A RP-HPLC and an U.V.single-or dual-wavelength spectrophotometrio methed for the determination of β-Carotene in bloud were established.A liquid chromatog-graph fitted with a column (250x×4.6mm) containing Spherisorb ODS-2, 5μm equipped with Tracor 951 .pump and a SPD-MIA variable wavevelongth UV-detector was used.The mobile phase is a mix of CH3OH,CHCl3 and 10% methanolto solution of (C2H5)3N(100:16:0.010).Amiodarone is used as an internal standard.The retention times for β-Carotene, α-Carotene and internal standard were12.84,13.85 and 8.77min respectively. The limit of detection was 5ng and the minimal detectable concentration was 25 ng/ml in plasma. A linear relationship of β-Carotene over the range from 25ng/ml to 2 000ng/ml, with r=0.999 8 was established; The within-day recovery and CT% were 101.2% and 3.1% (0.5μg/ ml; n=4), and those of between-days were 98.4% and 3.4 respectively. A model of UV-3000 Spectrophotometer was used in determining the absorption spectra ofβ-Carotene in petroleumether and in plasm.The quantitation of β-Carotene was harried out also with a single and dual-wavelength UV method. The correlation between the methods of HPLC and UVwas quite good, with r>0.9 and slope 1.0.