Abstract
An isocratic, paired-ion reversed-phase high-performance liquid chromatographic method for the determination of streptomycin and dihydrostreptomycin has been developed. The method employs a microparticulate reversed-phase column (μBondapak C18, LiChrosorb RP-18 or Ultrasphere Ion Pair), and a mobile phase composed of 0.02 M sodium hexanesulfonate and 0.025 M tribasic sodium phosphate in acetonitrile-water (8:92, v/v) at pH 6.0 with detection by ultraviolet absorbance at 195 nm. Resolution and simultaneous quantitation of streptomycin A, dihydrostreptomycin A, streptomycin B, streptidine and process-related substances can be achieved in less than 25 min, with a relative standard deviation of ca. 1.1% for the assay of either streptomycin sulfate or dihydrostreptomycin sulfate. The chromatographic bioequivalency data for streptomycin sulfate and dihydrostreptomycin sulfate were statistically identical with results obtained by the officially recognized microbiological assay. The method is designed for applicability to the analysis of other aminoglycoside antibiotics and antituberculous agents.
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