A novel and sensitive method for determining vitamin B3 and B7 by pre-column derivatization and high-performance liquid chromatography method with fluorescence detection

A new labeling reagent for vitamin analysis, 2-amino-10-ethyl acridine ketone (AEAO), has been synthesized and successfully applied to the analysis of vitamin B3 and vitamin B7 in different tea samples. The reaction of AEAO with vitamins could proceed easily and quickly in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) as condensing reagent within 45 min. The derivatives exhibited excellent fluorescence property with excitation and emission wavelengths of 290 nm and 430 nm, respectively. Response surface methodology (RSM) was applied to the optimization of pre-column derivatization. Solid phase extraction with HLB cartridges was used for the extraction and purification of water-soluble vitamins in tea samples. The LODs for vitamin B3 and vitamin B7 were 2.56 and 2.22 ng mL-1, respectively. The proposed method was successfully applied to the analysis of vitamin B3 and vitamin B7 in different tea samples. The study provided a highly sensitive method for accurate analysis of trace vitamins from natural products.