A generic method for the quantitative analysis of aminoglycosides (and spectinomycin) in animal tissue using methylated internal standards and liquid chromatography tandem mass spectrometry.
Aminoglycosides (AGs) are a large and diverse group of antibiotics. Although AGs may cause side effects of nephrotoxicity and ototoxicity, they are still occasionally being used for the treatment of serious infections. In this study the development of a method is described for the quantitative determination and confirmation of seven aminoglycosides (and relevant isomers) and spectinomycin in animal tissues. The extraction was based on an extraction followed by a concentration and clean-up step using weak cation exchange solid phase extraction. The separation was performed by ion-pair liquid chromatography on a C(18) column followed by mass spectrometric detection. The method was validated according to the EU requirements for a quantitative confirmatory method. Permethylated aminoglycosides (in-house synthesised internal standards) were used for accurate quantification. The accuracy of the analyses of AGs in kidney ranged from 94 to 111%, intra-day precision ranged between 2.5 and 7.4% (R.S.D.(r)) and inter-day precision ranged between 2.2 and 17.3% (R.S.D.(RL), n=21, MRL level). Accuracy (muscle tissue) varied from 83 to 128% with an intra-day precision between 2.2 and 17.3% (R.S.D.(r), n=7, MRL level). From the results it was concluded that the method was able to monitor MRL levels which ranged from 750 to 20,000 microgkg(-1) for kidney and from 50 to 10,000 microgkg(-1) for muscle tissue.
van Holthoon
Frédérique L
FL
RIKILT - Institute of Food Safety, Wageningen UR, Bornsesteeg 45, 6708 PD, P.O. Box 230, 6700 AE Wageningen, The Netherlands.
Essers
Martien L
ML
Mulder
Patrick J
PJ
Stead
Sara L
SL
Caldow
Marianne
M
Ashwin
Helen M
HM
Sharman
Matthew
M
D000617:Aminoglycosides / Q000032:analysis* / Q000302:isolation & purification; D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004350:Drug Residues / Q000032:analysis* / Q000302:isolation & purification; D012015:Reference Standards; D015203:Reproducibility of Results; D000198:Spectinomycin / Q000032:analysis* / Q000302:isolation & purification; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D000198:Spectinomycin