Micro determination of tobramycin in serum by high-performance liquid chromatography.

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Abstract
A rapid, simple, and accurate method for the determination of tobramycin in serum by high-performance liquid chromatography (HPLC) has been developed. The method is sensitive to 0.2 μg/ml using only 20 μl of serum. The serum proteins are precipitated with methanol. To the supernatant, a counter-ion reagent is added, then an aliquot of the solution is injected into the chromatograph. The determination of tobramycin is performed by a combination of reverse-phase, ion-pair chromatography, post-column derivatization with o-phthalaldehyde, and fluorescence detection. Comparisons with a microbiological assay and with a homogeneous enzyme immunoassay gave a correlation coefficient of 0.941 and that of 0.978, respectively.

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