Simultaneous Determination of B1, B2, B3, B6, B9, and B12 Vitamins in Premix and Fortified Flour Using HPLC/DAD: Effect of Detection Wavelength

Simultaneous determination of water-soluble B vitamins is a troublesome analytical procedure since they have greatly variable structures and acid-base properties which imposed difficulties on eluting them in short time selecting wavelength detection. The aim the present study was to develop simple method that overcomes these difficulties. successful simultaneous B1, B2, B3, B6, B9, B12 premix fortified flour by extracting with 0.1% (w/v) ascorbate ethylene diamine tetra-acetic acid (EDTA) solution followed using gradient mobile phase consisting 0.03% trifluoroacetic aqueous (pH 2.6) acetonitrile high performance liquid chromatography (HPLC) instrument coupled diode array detector (DAD). Elution completed within 9.3 min, lowest values obtained for limit quantification (LOQ) were 0.6, 0.2, 0.8, 0.3, 0.5, 0.7 μg/mL, respectively, at four wavelengths 361, 280, 265, 210 nm. In general, variation detection range from 361 nm affects sensitivity but had marginal effect linearity LOQ developed its application determining flour. exhibited highest though resulted higher respect Noteworthy, B2 relatively RSD compared lower wavelengths. Thus, 265 280 may be more favorable over reported work does not require any sample cleanup/preconcentration steps, chromatographic elution achieved min without need ion-pairing reagents.