Clinical Chemistry
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Clinical Chemistry 25: 1222-1225, 1979;
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Clinical Chemistry, Vol 25, 1222-1225, Copyright © 1979 by American Association for Clinical Chemistry

Chemical assay, involving liquid chromatography, for aminoglycoside antibiotics in serum

SE Back, I Nilsson-Ehle and P Nilsson-Ehle

We describe a chemical assay involving "high-pressure" liquid chromatography for the quantitative determination of three aminoglycosides: netilmicin, tobramycin, and gentamicin. The drugs are separated from serum by means of precipitation of the serum proteins with acetonitrile after dilution with a buffer. The aminoglycosides are quantitatively extracted into the supernate, which is further purified by a two-step partition procedure involving derivatization of the drugs with o-phthalaldehyde. The drug derivatives are separated by reversed- phase chromatography and detected by on-line fluorometry. Sensitivity is 1 mg/L for tobramycin and 0.5 mg/L for netilmicin and gentamicin. Intra- and interassay variation was below 8%. Analytical recovery of each of the three drugs was 92 to 100%. Correlation with microbiological and radioimmunological assay methods was good. The assay is rapid (about 30 min), precise, and specific, and seems suitable for use in a routine clinical laboratory.




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Copyright © 1979 by the American Association for Clinical Chemistry.