Rapid determination of vitamin B 2 and B 12 in human urine by isocratic liquid chromatography

Abstract

A simple and rapid method for the identification and quantification of vitamin B 2 and B 12 in human urine has been developed using reverse phase high performance liquid chromatography (HPLC) and the peaks identity were confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). HPLC separation was performed in single wavelength detector ( λ 365) mode and separated isocratically using mobile phase methanol: 1 mM aqueous TFA (1:4) in C18 column. The calibration graphs plotted with different concentrations of vitamin B 2 and B 12 was linear with a correlation coefficients ( r 2) = 0.9975 and 0.9985, respectively. The recoveries of vitamin B 2 and B 12 were above 87% and 90%, respectively. The results of this present study suggest that the proposed method may be simple and convenient way of identifying and quantifying vitamin B 2 and B 12 from human urine.