The I5N NMR spectra of some of the components of the nebramycin complex are reported. Resonances are assigned through comparisons of spectra of the individual factors. The observed chemical shifts are broadly consistent with the current knowledge of substituent effects in I5N NMR spectroscopy. Titration curves can be used to obtain pK, values for each of the nitrogens of a compound. The results of this study were used to confirm the identification of a monoacetyl derivative of tobramycin.