Determination of vitamin B₁ in seawater and microalgal fermentation media by high-performance liquid chromatography with fluorescence detection
2005
He, Hong-Zhi | Li, Hua-Bin | Chen, Feng
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 mmx4.6 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 5.00x10-¹⁰ mol L-¹ to 5.00x10-⁷ mol L-¹ for vitamin B₁ with a correlation coefficient of 0.9991 (n=9). The detection limit was 1.0x10-¹⁰ 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%.
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