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Determination of diminazene aceturate in pharmaceutical formulations by HPLC and identification of related substances by lC/MS

Atsriku, C. and Watson, D.G. and Tettey, J.N.A. and Grant, M.H. and Skellern, G.G. (2002) Determination of diminazene aceturate in pharmaceutical formulations by HPLC and identification of related substances by lC/MS. Journal of Pharmaceutical and Biomedical Analysis, 30 (4). pp. 979-986. ISSN 0731-7085

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Abstract

A validated, reversed-phase, isocratic high-performance liquid chromatographic method for the simultaneous assay of diminazene aceturate, antipyrine (excipient) and diminazene impurities in pharmaceutical formulations is described. The chromatographic system consisted of a Lichrospher-60 RP-select B column with a mobile phase composition of acetonitrile–methanol–ammonium formate (pH 4.0, 20 mM) (10:10: 80 v/v/v) and UV detection at 254 nm. The method is specific, precise and accurate for the determination of diminazene in the presence of its manufacturing and degradation impurities with a limit of detection and quantification of 50 ng/ml and 10 μg/ml (RSD<3.0%), respectively. The major manufacturing impurity [1-(4 amidino phenyl)3-(4 carbamoyl phenyl)-triazene] and a degradant (p-aminobenzamidine) of diminazene aceturate have been resolved and identified by liquid chromatography/electrospray ionization-mass spectrometry operated in a positive ion mode.