Simultaneous determination of vitamin A and E in microamount of human serum by dispersive liquid phase micro-extraction – high performance liquid chromatography
ZHENG Fengjia, BIE Mingjiang, SUN Chengjun*
West China School of Public Health, Sichuan University, Chengdu 610041, China
Abstract:[Objective]To establish a dispersive liquid phase microextraction–high performance liquid chromatography for simultaneous determination of vitamin A and E in microamout of human serum.[Method]Twenty microliter of human serum was taken and 50μl methanol added to precipitate the protein, and 50μl chloroform added to extract vitamin A and E ultrasonically. After centrifuged at 12000rpm for 5 min, the lower chloroform layer was taken and injected for HPLC analysis. The optimized chromatographic conditions were as follows: analytical column: (Eclipse XDB-C8, 150 mm × 4.6 mm，5μm); mobile phase: methanol-water (96+4); flow rate: 0.80 ml/min; column temperature: 25℃; detection wavelength: 325nm for vitamin A and 292nm for vitamin E, respectively.[Results]The correlation coefficients for the standard curves were greater than 0.999 for both vitamin A and E; the detection limits of the method were 0.035μg ∕ ml for vitamin A and 0.09μg ∕ ml for vitamin E. The recoveries of vitamin A and E in human serum ranged from 90.4%～103.2% and 81.0%～92.1%, respectively.[Conclusion]The method is sensitive, accurate, rapid, simple, and suitable for the determination of vitamins A and E in microamount of human serum.