Spectrophotometric Determination of Pyridoxine Hydrochlorine in Pharmaceutical Preparations and Foods

An easy and sensitive spectrophotometric assay of pyridoxine is described. The procedure is based on formation of an azo dye by the reaction of pyridoxine with diazotized 2,4-dinitroaniline followed by the reaction of the dye with Hg2+ ions to form a stable complex with maximum absorbance at 545 nm. The system obeys Beer's law for 4-75 μg pyridoxine hydrochloride in an overall aqueous volume of 25 mL (correlation coefficient, 0.9998). On extraction into 5 mL butan-1-ol, the system obeys Beer's law in the range 0.8-15 μg pyridoxine hydrochloride at 545 nm. The color is stable for 60 min in both aqueous and organic phases (molar absorptivity, 3.7 × 104 L/mol · cm; coefficient of variation, 3.1%, n = 10). The pyridoxine contents of pharmaceutical preparations, a processed foodstuff, and 2 rice samples were determined by using the proposed method. Assay reliability was established by recovery studies and parallel determination using a reported method.