Quantitative determination and separation of analogues of aminoglycoside antibiotics by high-performance liquid...

Commercial bulk products and pharmaceutical drug formulations of aminoglycoside antibiotics obtained by fermentation (kanamycin, gentamicin, sisomicin and tobramycin) or by synthesis (amikacin) were analysed with high-performance liquid chromatography on a C8 reversed-phase column. The method is based on a pre-column derivatization of the aminoglycosides with a 2,4,6-trinitrobenzenesulphonic acid reagent and UV detection (350 nm). The quantitative determination was carried out vs. an external standard; both peak heights and areas were used. A gentamicin mixture was separated into five or four components, depending on the column used. Amikacin was separated from its possible regioisomers and kanamycin A was easily separated from its minor components B and C.