Validation of a method for the determination of aminoglycosides in different matrices and species based on an in-house concept

Abstract

A simple, rapid, and sensitive method for the determination and confirmation of the aminoglycosides streptomycin, dihydrostreptomycin, spectinomycin, apramycin, kanamycin, paromomycin, gentamicin and neomycin in cow’s milk as well as in bovine and porcine muscle and kidney was developed. Validation was performed on the basis of an in-house concept with different factor-level combinations in accordance with Commission Decision 2002/657/EC. After extraction with trichloroacetic acid solution, clean-up was performed by way of SPE. LC-MS/MS analysis was carried out by means of an HILIC column for the separation of the analytes, and by using MS/MS in positive ESI mode to measure the transitions of the substances in MRM mode. For quantification, matrix calibration curves in the linear range around the MRLs as well as the internal standard tobramycin were used. The calculated validation parameters like CCα, CCβ, recovery (94–103%), relative repeatability RSD_r (3.6–9.7%), and relative within-laboratory reproducibility RSD_wR (4.6–10.0%) fulfilled the requirements of Commission Decision 2002/657/EC.

Keywords:

• veterinary drug residues
• antibiotics
• tissue and milk
• LC-MS/MS
• clean-up
• extraction
• method validation

Acknowledgements

Special thanks to Ms S. Mönch, Ms M. Jüsgen, Ms K. Kindt and Ms C. Lund for their assistance and laboratory work.