DETERMINATION OF VITAMIN-E IN AQUATIC ORGANISMS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION

A liquid chromatographic (HPLC) method has been developed for the quan titative determination of different forms of vitamin E (alpha-, gamma- , and delta-tocopherol) in aquatic organisms. The assay consists of a simple extraction with methanol containing butylhydroxytoluene (BHT) a s an antioxidant, followed by reversed-phase chromatography with fluor escence detection. The efficiency of the extraction method was equival ent or superior to that of more complex approaches for the isolation o f tocopherols. Linearity has been achieved over the range of 0.02 to 3 mu g/ml for alpha-tocopherol and within-run and between-run coefficie nts of variation at three levels were 0.7-2.9 and 1.2-3.7%, respective ly. The recovery at three concentrations ranged from 73.8 to 96.6% and the minimal quantity that could be detected was 0.6 ng. Comparable fi gures were obtained for gamma- and delta-tocopherol. This method has b een routinely applied to determine vitamin E in Artemia, rotifers, tur bot and sea bass larvae, and shrimp postlarvae. (C) 1996 Academic Pres s, Inc.