Determination of vitamin B6 in pharmaceutical formulations by flow injection-solid phase spectrophotometry

In this work, a new solid phase spectrophotometric method in association with flow injection analysis for Vitamin B6 (pyridoxine) determination has been developed with direct measurement of light-absorption in C18 material. In the developed method, successive passage of the complex, previously formed in the flowing stream, and eluent through the flow cell and continuous monitoring of the process provided the analytical information needed to determine pyridoxine. Pharmaceutical samples containing Vitamin B6 were previously dissolved in 0.1 mol l(-1) phosphate buffer solution (pH 7.5) and a sample volume of 235 microl was injected directly into carrier stream consisting of a mixture of methanol and 0.1 mol l(-1) phosphate buffer solution adjusted to pH 7.0 (1+1, v/v). The blue indophenol dye produced from the reaction between pyridoxine and N,N-diethyl-p-phenylenediamine after oxidation by potassium hexacyanoferrate(III) was quantitatively retained on C18 support and the spectrophotometric detection was performed simultaneously at 633 nm. The retained complex was quickly eluted from C18 material with the eluent stream consisting of a mixture of methanol and 0.01 mol l(-1) HCl (6+4, v/v). The results showed that the proposed method is simple, rapid and the analytical response is linear in the concentration range of 0.5-10 and 0.2-4 mg l(-1) using 235 and 860 microl of sample, respectively. The limits of detection are 0.15 and 0.060 mg l(-1) and the R.S.D. are 3.6% (at 2 mg l(-1) level) and 4.0% (at 1 mg l(-1) level) using sample volume of 235 and 860 microl, respectively. The system presented an analytical throughput of 15 determinations per hour when a sample volume of 235 microl was utilized. The procedure was successfully applied to the determination of Vitamin B6 in pharmaceutical formulations containing vitamins of B group and others active principles such as Vitamin C and minerals.