Determination of vitamin B1 in seawater and microalgal fermentation media by high-performance liquid chromatography with fluorescence detection
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- He, HZ., Li, HB. & Chen, F. Anal Bioanal Chem (2005) 383: 875. doi:10.1007/s00216-005-0062-1
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A highly sensitive high-performance liquid chromatographic method with fluorescence detection has been developed for determination of vitamin B1. Vitamin B1 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 B1 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 B1 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%.