This research involves the development of spectrophotometric method for determination of pyridoxine hydrochloride by oxidative coupling reaction with N,N-dimethylparaphenylenediamine dihydrochloride (DMPPDAD ( in a basic medium (pH 11.6) in the presence of potassium ferricyanide to produce an intense green colour, soluble in water, stable product and absorbs at 650 nm. Beer's law was in the linear range 6-30 μg/ml of pyridoxine hydrochloride, the molar extinction coefficint, Sandellʼs sensitivity index and detection limit were 4.338 × 103 L. mol-1.cm-1 , 0.0474 μg.cm-2 and 0.756 μg/ml respectively. The RSD value was 0.63 - 1.41% depending on the concentration. This method was applied successfully to the determination of pyridoxine hydrochloride in pharmaceutical preparation samavite B6 tablet with recovery of not less than 99.5%.