Serum gentamicin, netilmicin and tobramycin assays by high performance liquid chromatography

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Abstract
An HPLC method for the routine determination in serum of gentamicin, tobramycin and netilmicin is described. The technique involves the extraction of the aminoglycosides from serum onto Amberlite CG50 resin and the formation of fluorescent derivatives with O-phthaldehyde. The derivatives were separated on a reverse phase Spherisorb 5-ODS column, whereby gentamicin gave four distinct peaks Cl, Cla, C2a and C2, whereas tobramycin and netilmicin gave a single peak. Comparison of the gentamicin assay with a microbiological assay technique and with the PHLS National Quality Assessment Scheme along with its good sensitivity and reproducibility, indicate it to be suitable for routine use in the laboratory.