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Updated: Aug 4, 2026
Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Liquid chromatographic determination of thiamine in rodent feed by postcolumn derivatization and fluorescence
T A Gehring1, W M Cooper, C L Holder
1U.S. Food and Drug Administration, National Center for Toxicological Research, Division of Chemistry, Jefferson, AR 72079, USA.
Abstract:
A liquid chromatographic method was developed for determination of the essential nutrient thiamine (vitamin B1) in rodent feed. Thiamine was extracted with hydrochloric acid, separated by reversed-phase liquid chromatography, derivatized postcolumn to thiochrome with potassium hydroxide and potassium ferricyanide, and detected by fluorescence. Excitation and emission wavelengths were 370 and 430 nm, respectively. Detector response was linear in the range of 2.58 to 15.5 ng of thiamine injected. Instrument detection limit was 5 pg of thiamine injected.
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