An LC Ion-Pairing Method for the Determination of Fe(II) in Ferrous Bisglycinate Pharmaceutical Formulation

A reversed-phase ion-pairing liquid chromatographic method was developed and validated for the assay of Fe(II) in ferrous bisglycinate (Fe-bis-gly) capsules using 4-(2-pyridylazo) resorcinol reagent. The analysis was carried out using a Gemini RP-18 (150 mm x 4.6 mm I.D., particle size 5 mu m) analytical column; the mobile phase consisted of a mixture of acetonitrile-water (28:72 v/v) containing 1 mM tetrabutylammonium hydrogensulfate and 1% phosphate buffer (pH 8.0). The flow rate was 1.0 mL min(-1) and the detection was achieved with a photodiode array (PDA) detector at 706 nm. The specificity of the method was proved using stress conditions and evaluated using a PDA detector. The data validation showed that the method is specific, fast, accurate, and reproducible for the determination of Fe-bis-gly in dosage form. The response was linear over a range of 1.0-2.6 mu g mL(-1) (r = 0.9999). The accuracy of the method ranged from 98.02 to 102.75%. The RSD values for intra- and inter-day precision studies were below 1.3 and 1.1%, respectively. There was no interference of the excipients on the determination of the active pharmaceutical ingredient.