A simple and sensitive spectrofluorimetric method has been developed and validated for determination of amikacin sulfate, neomycin sulfate and tobramycin in pure forms, pharmaceutical formulations and human plasma. The method was based on condensation reaction of cited drugs with ninhydrin and phenylacetaldehyde in buffered medium (pH 6) resulting in formation of fluorescent products which exhibit excitation and emission maxima at 395 and 470nm, respectively. The different experimental parameters affecting the development and stability of the reaction products were carefully studied and optimized. The calibration plots were constructed with good correlation coefficients (0.9993 for tobramycin and 0.9996 for both neomycin and amikacin). The proposed method was successfully applied for the analysis of cited drugs in dosage forms with high accuracy (98.33–101.7)±(0.80–1.26)%. The results show an excellent agreement with the reference method, indicating no significant difference in accuracy and precision. Due to its high sensitivity, the proposed method was applied successfully for determination of amikacin in real human plasma.
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