Abstract
Reported differences in the order of elution of fluorescent o-phthalaldehyde derivatives of gentamicins C 1, C 1a and C 2 on apparently similar reversed-phase HPLC systems are explained. The differences arise from the relatively higher sensitivity of the κ value of the gentamicin C 1 derivative to the presence of cations in the eluting solvent. This effect also depends on the commercial source of the bonded ODS phase used and is inversely related to the surface coverage of the silica with alkyl chains. The evidence suggests that this phenomenon results from an interaction between the residual silanol groups on the column support and secondary amino groups on the gentamicin derivatives, the C 1 derivative differing from the C 1a and C 2 derivatives in having two rather than only one such group.
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