Abstract
Pimaricin (PMR) is a polyene antifungal drug commonly used as the primary medicine for treating fungal keratitis. In this study, a selective and novel spectrofluorimetric method was designed, optimized, and validated to determine pimaricin in its ophthalmic preparations. The method based on the formation of a nitrobenzoxadiazole-based yellow fluorophore upon the reaction of pimaricin with 4-chloro-7-nitro-1,2,3-benzoxadiazole (NBD-Cl). The fluorescence intensity of the fluorophore was measured at the emission wavelength of 538.5 nm (excitation = 473.5 nm). The factors affecting the reaction (reagent volume, alkalinity, reaction temperature, heating time, volume and type of buffer solution, and the dilution solvent) were investigated and optimized. The calculated linear range was 0.25–1.5 µg/mL, and the limits of detection and quantitation for the method were 0.047 and 0.143 µg/mL, respectively. The selectivity of the method for PMR in the presence of the pharmaceutical additive benzalkonium chloride was proved. The established method was successfully applied to determine PMR in its pharmaceutical ophthalmic preparation (Natamycin® ophthalmic drops) with an accuracy of 103.79 ± 1.83%. The proposed method is practical and valuable for its routine application in quality control laboratories for analysis of PMR in its ophthalmic pharmaceutical preparation.
Published Version
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