Determination of vitamin B 1 in seawater and microalgal fermentation media by high-performance liquid chromatography with fluorescence detection

Abstract

A highly sensitive high-performance liquid chromatographic method with fluorescence detection has been developed for determination of vitamin B 1. Vitamin B 1 was converted into a fluorescent compound by treatment with hydrogen peroxide-horseradish peroxidase and the derivative was subsequently analyzed by HPLC on a Waters Spherisorb ODS2 column (250 mm×4.6 mm ID, 5 μm) with 40:60 methanol-pH 8.5 acetate buffer solution as mobile phase and fluorescence detection at 440 nm (with excitation at 375 nm). The calibration graph was linear from 5.00×10 -10 mol L -1 to 5.00×10 -7 mol L -1 for vitamin B 1 with a correlation coefficient of 0.9991 (n=9). The detection limit was 1.0×10 -10 mol L -1. The method was successfully used for determination of vitamin B 1 at pg mL -1 levels in microalgal fermentation media and seawater after solid-phase extraction. Recovery was from 89 to 110% and the relative standard deviation was in the range 1.1 to 4.3%. © Springer-Verlag 2005.