An isocratic separation of underivatized gentamicin components, 1H NMR assignment and protonation pattern

Wojciech Lesniak, John Mc Laren, Wesley R. Harris, Vincent L. Pecoraro, Jochen Schacht

Research output: Contribution to journalArticlepeer-review


A simple method for the separation of the major components of commercial gentamicin sulfate (C-1, C-1a, C-2, C-2a) by high-performance liquid chromatography (HPLC) on an analytical and a semipreparative scale was developed. The method utilized ion-pair reversed-phase chromatography, isocratic elution with an aqueous solution containing 9% trifluroacetic acid and 2.5% acetonitrile as the mobile phase at a flow rate of 2 and 9 mL/min for analytical and semipreparative columns, respectively. Detection was carried out at 213 nm without derivatization. The protonation pattern of the separated gentamicins was determined by potentiometry and 15N and 1H NMR. The full proton NMR assignment for gentamicin C-1 was obtained through the use of 1H 1D and 2D 1H-1H COSY measurements.

Original languageEnglish (US)
Pages (from-to)2853-2862
Number of pages10
JournalCarbohydrate Research
Issue number24
StatePublished - Nov 21 2003
Externally publishedYes


  • Aminoglycosides
  • HPLC
  • NMR spectroscopy
  • Potentiometery

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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