title: Development and validation of a novel LC non-derivatization method for the determination of amikacin in pharmaceuticals based on evaporative light scattering detection.
abstract: A novel method for the direct determination of the aminoglycoside antibiotic amikacin and its precursor component kanamycin was developed and validated, based on reversed phase LC with evaporative light scattering detector (ELSD). ELSD response to amikacin was found to be enhanced by: (a) use of ion-pairing acidic reagents of increased molecular mass, (b) increase of mobile phase volatility and (c) decrease of peak width and asymmetry (obtained by controlling the mobile phase acidity and/or ratio of organic solvent to water). Utilizing a Thermo Hypersil BetaBasic C(18) column, the selected optimized mobile phase was water-methanol (60:40, v/v), containing 3.0 mll(-1) nonafluoropentanoic acid (18.2mM) (isocratic elution with flow rate of 1.0 mlmin(-1)). ELSD experimental parameters were: nitrogen pressure 3.5 bar, evaporation temperature 50 degrees C, and gain 11. Amikacin was eluted at 8.6 min and kanamycin at 10.4 min with a resolution of 1.5. Logarithmic calibration curves were obtained from 7 to 77 microgml(-1) (r>0.9995) for amikacin and 8 to 105 microgml(-1) (r>0.998) for kanamycin, with a LOD equal to 2.2 and 2.5 microgml(-1), respectively. In amikacin sulfate pharmaceutical raw materials, the simultaneous determination of sulfate (t(R)=2.3 min, LOD=1.8 microgml(-1), range 5-40 microgml(-1), %R.S.D.=1.1, r>0.9997), kanamycin and amikacin was feasible. No significant difference was found between the results of the developed LC-ELSD method and those of reference methods, while the mean recovery of kanamycin from spiked samples (0.5%, w/w) was 97.3% (%R.S.D.<or=2.0, n=6). Further, the developed method was applied for the determination of amikacin in pharmaceutical formulations (injection solutions) without any interference from the matrix (recovery from spiked samples ranged from 95.6 to 103.8%).
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Panepistimiopolis, Athens 15771, Greece.
authors.forename: Evagelia G
authors.initials: EG
authors.lastname: Galanakis
authors.forename: Nikolaos C
authors.initials: NC
authors.lastname: Megoulas
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
authors.forename: Michael A
authors.initials: MA
authors.lastname: Koupparis
mesh_terms: D000465:Algorithms; D000583:Amikacin / Q000032:analysis*; D000900:Anti-Bacterial Agents / Q000032:analysis*; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D007612:Kanamycin / Q000032:analysis; D008027:Light; D008970:Molecular Weight; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D013431:Sulfates
chemical_list: D000900:Anti-Bacterial Agents; D007202:Indicators and Reagents; D013431:Sulfates; D007612:Kanamycin; D000583:Amikacin