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In order to develop an analytical method for amikacin and butirosin in presence of their parent antibiotics, kanamycin A and ribostamycin, high-performance liquid chromatographic technique and microbioassay method were evaluated and compared. Using high performance liquid chromatography, two acylated antibiotics, amikacin and butirosin was partially separated from their parent antibiotics, to provide a qualitative analytical method. In microbioassay using Pseudomonas aeruginosa TI-13, a producer of aminoglycoside-3’’-phosphotransferase I, only acylated antibiotics were selectively analyzed when paper disc-susceptibility assay was used. The standard curve showed a good correlation between the response and dose in semilogarithmic plot with correlation coefficients above 0.96, and analytical deviation from expected dose was within 10%.
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