Identification of gentamicin impurities by liquid chromatography tandem mass spectrometry.
An HPLC/MS/MS method was developed for identification of impurities in gentamicin. The HPLC was performed on a Synergy Hydro-RP column using 50 mM trifluoroacetic acid (TFA), pH 2 adjusted with ammonium solution and methanol as mobile phase. All impurities in gentamicin were separated from main gentamicin components. Atmospheric pressure chemical ionization (APCI) was used and product mass spectra of protonated molecules were acquired. Seventeen impurities were detected in gentamicin. Reference compounds: gentamicins: C2b, B, B1, G-418, sisomicin, garamine and gentamines: C1, C1a, C2, C2a were used for spectra interpretation and impurities identification. All MS/MS spectra were interpreted and fragmentation transitions for gentamicins and in general for aminoglycoside antibiotics (AG) were proposed. All impurities were identified. More than one isomere were proposed for three impurities.
Grahek
Rok
R
Lek Pharmaceuticals d.d., Sandoz Development Center Slovenia, Verovskova 57, SI-1526 Ljubljana, Slovenia. rok.grahek@sandoz.com
Zupancic-Kralj
Lucija
L
D000617:Aminoglycosides / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D004340:Drug Contamination; D005839:Gentamicins / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007477:Ions; D008956:Models, Chemical; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014269:Trifluoroacetic Acid / Q000737:chemistry
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D005839:Gentamicins; D007477:Ions; D000644:Quaternary Ammonium Compounds; D014269:Trifluoroacetic Acid