Gas-liquid chromatographic method for the assay of aminoglycoside antibiotics in serum

Abstract

A gas-liquid chromatographic (GLC) method is presented for the rapid analysis of gentamicin, tobramycin, netilmicin, and amikacin from human serum. This procedure may have application to all aminoglycoside drugs. The three isomers of gentamicin are resolved as two bands, while tobramycin, netilmicin, and amikacin appear in this system as single bands. Normal serum constituents do not interfere with chromatograms. Thus far, no assay interference has been found in cases where other drugs and antibiotics were administered concurrently with aminoglycoside therapy. Dose-response data demonstrating linear recovery are included for all four aminoglycosides as well as a comparison of the GLC method with the microbiological method for the assay of gentamicin and amikacin. Quantitation is based upon the relative response of the antibiotics to a fixed amount of the internal standards, either kanamycin A or paromomycin B. These standards are clearly resolved as symmetrical peaks from the antibiotics of assay interest. Isothermal chromatographic analysis time is less than 8 min, while total assay time per single serum specimen is approximately 50 min. Preparation of serum includes: precipitation, evaporative drying of the supernatant, a two-stage derivatization (N-trimethylsilylimidazole, N-heptafluorobutyrylimidazole), and a single hexane extraction with a water wash. The methodology described may be applied to the analysis of other compounds (e.g., saccharides, amino-saccharides, amino acids, etc.) which do not rapidly partition into an organic phase.