DEVELOPMENT AND VALIDATION OF A 96-MICROWELL BASED ASSAY OF AZITHROMYCIN BY CHARGE TRANSFER COMPLEXATION WITH 2,3- DICHLORO-5,6-DICYANO-1,4-BENZOQUINONE

Author: Ejiofor Innocentmary Ifedibaluchukwu
Department: Pharmaceutical and Medicinal Chemistry
Affiliation: Nnamdi Azikiwe University Awka

A 96-microwell-based visible spectrophotometric assay was developed and validated for the determination of azithromycin in tablets. The formation of a coloured charge-transfer complex between azithromycin as n-electron donor, and 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone, as π-electron acceptor, was investigated, and employed as a basis for the development of the proposed assay. The proposed assay was carried out in a 96-microwell plate. The absorbance of the coloured charge transfer complex was measured at 490 nm wavelength using a microwellplate absorbance reader. The optimum conditions of the reaction and the analytical procedures of the assay were established. Under the optimum conditions, linear relationship with good correlation coefficient was found between the absorbance and the concentration of azithromycin in the range of 0.076 – 0.180 mg/ml, with the equation y=1.048x and R2 = 0.993. The ΔG at different temperatures were found to be -8429.8, -9342.6, -13011.9, -12678, and -15124.8 Kcal/mol. The limits of detection and quantitation were found to be 0.023 and 0.069 mg/ml respectively. The procedure gave good precision. The assay was applied successfully to the analysis of azithromycin in tablets with satisfactory result. The assay described has high throughput property, consumes little volume of organic solvent, offers reduction in analysis cost and the exposure of the analyst to the toxic effects of organic solvent.

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