Determination of Vitamin B12 Using Differential Pulse Polarography

Abstract

Vitamin B12 is a type of vitamin also known by the name cobalamin. B12 is involved in many metabolism activities, including DNA synthesis, nervous system, red blood formation and immune system. Therefore, we chose the Differential Pulse Polarography (DPP) method is that has a high sensitivity for the determination of vitamin B12. This determination was possible with cobalt present in vitamin B12 structure. Since Co(III) is formed from the oxidation of the vitamin, its polarographic behavior had to be determined in various electrolytes such as acetate, borate, phosphate and ammonia. The polarograms of Co(III) were taken in these electrolytes in which 1.0 M NH3/ (pH = 9.8) and 1.0 M AcOH/AcO- (pH = 4.0) were found as the most suitable electrolytes. This method was successfully applied vitamin of B12 determination in a 1 mL ampoule with high precision. The LOD was found as 3.7 × 10-7 for instead of (S/N = 3). Besides Co(III), interference effects of Zn(II), Ni(II), Cr(III), Fe(III), Cu(II), Cd(II) and Se(IV) were also studied. It was found that only Zn(II) peak had an overlap Co(III) peak in ammonium buffer. This problem could be solved by working in 1.0 M AcOH/AcO- (pH = 4.0) buffer. B12, which is 1000 μg in 1 mL vitamin ampoule, was found for 4 measurements as 999 ± 15 μg as a result of 95% confidence interval.