Control of the composition of gentamicin sulphate by proton magnetic resonance spectrometry.

The proportions of the main components present in gentamicin sulphate complex, gentamicins C1, C1a and C2, can be monitored by 1H nuclear magnetic resonance (nmr) spectrometry. The method depends on measurement of the peak heights of signals for N-methyl and C-methyl groups present in all three components and of those present in C1 and C2 only, followed by calculation of peak height ratios to control composition within acceptable limits. The precision and reproducibility of the method have been established through two collaborative studies each involving seven laboratories. In the second study, with an improved procedure, the mean variance between laboratories with 10 samples was 3.4 X 10(-4) for the N-methyl ratio of the peak at delta 2.75 to that at delta 2.95, and 1.25 X 10(-3) for the C-methyl ratio of the peak at delta 1.25 to that at delta 1.35. Within laboratories the mean variance for triplicate determinations was 7.4 X 10(-5) and 8.9 X 10(-5) respectively. The data presented here form the experimental basis for the test controlling the composition of gentamicin sulphate in the British Pharmacopoeia 1973: Addendum 1975, and for the introduction into the British Pharmacopoeia of nmr spectrometry as an analytical technique. The reference standards and all batches of gentamicin sulphate intended for therapeutic use in the United Kingdom examined by this procedure comply with the limits laid down.