Determination of nitrofuran metabolites in milk by liquid chromatography-electrospray ionization tandem mass spectrometry.
An LC-ESI-MS-MS method for the analysis of metabolites of four nitrofurans (furazolidone, furaltadone, nitrofurazone and nitrofurantoin) in raw milk has been developed. The samples were achieved by hydrolysis of the protein-bound drug metabolites, derivatization with 2-nitrobenzaldehyd (2-NBA) and clean-up extraction liquid-liquid with ethyl acetate. LC separation was achieved by using a Phenomenex Luna C-18 column. The mass spectrometer operated in multiple reaction monitoring mode (MRM) with positive electro-spray interface (ESI). The method validation was done according to the criteria laid down in Commission Decision No. 2002/657 EC. The validation includes the determination of linearity, repeatability, within-laboratory reproducibility, accuracy, decision limit (CCalpha) and detection capability (CCbeta). The calibration curves were linear, with typical (R(2)) values higher than 0.991. The coefficient of variation (CV, %) was lower than 9.3% and the accuracy (RE, %) ranged from -9.0% to 7.0%. CV within-laboratory reproducibility was lower than 13%. The limits of decision (CCalpha) and detection capability (CCbeta) were 0.12-0.29 microg/kg and 0.15-0.37 microg/kg, thus below the minimum required performance limit (MRPL) set at 1 microg/kg by the UE. This validated method was successfully applied for the determination of nitrofuran metabolites in a large number of milk samples.
Rodziewicz
Lech
L
Department of Hygiene Veterinary, ul. Zwyciestwa 26a, 15-959 Białystok, Poland. rodziewicz@wiw.bianet.com.pl <rodziewicz@wiw.bianet.com.pl>
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D008892:Milk / Q000737:chemistry*; D009581:Nitrofurans / Q000032:analysis*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D019155:Veterinary Drugs / Q000032:analysis*
D009581:Nitrofurans; D019155:Veterinary Drugs