ANALYSIS OF HUMAN SERUM TOCOPHEROL ISOMERS BY NORMAL-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC PROCEDURE / 营养学报

ABSTRACT

Objective: A high-performance liquid chromatographic (HPLC) procedure with a fluorescence detector was developed to rapidly separate ?,?,?,?- tocopherol isomers in human serum Methods: The HPLC system consisted of Inertsil silica column (100-A, 3?m,4.6mm?250mm) and 7% (v/v) methyl-tert- butyl ether in n-hexane as mobile phase . Prior to HPLC, the serum sample wa deproteined by ethanol (BHT 0.0625%) and the tocopherol isomers were efficiently extracted in thei original isomeric conformations using n-hexane-ethyl acetate (51) in the presence of 2,6-bi-buty p-methylphenol (BHT). Result: The quantification limits, defined as the lowest quantitatively measurable concentration of the different compounds (ng/ml) are calculated according to the experiment?-tocophero 1.0,?-tocopherol 1.0,?-tocopherol 0.5,?-tocopherol 0.5. The recovery rates are between 95%~105% Correlation coefficients are over 0.999 when the concentration is between 5 ng/ml~5 ?g/ml. Conclusion This technique is suitable for assay of tocopherol isomers in human serum at all ages.