Confirmatory method for the determination of resorcylic acid lactones in urine sample using immunoaffinity cleanup and liquid chromatography-tandem mass spectrometry.
The presence of zeranol (alpha-zearalanol) in urine samples due to natural contamination or illegal treatment is under debate within the European Union. The simultaneous determination of zeranol, its epimer taleranol (beta-zearalanol), zearalanone and the structurally related mycotoxin zearalenone with the corresponding alpha- and beta-zearalenol metabolites appears to be critical in deciding whether an illegal use has occurred. The aim of this study is to develop and validate a simple analytical procedure applicable to bovine and swine urine samples for the determination of all six resorcylic acid lactones. After an enzymatic deconjugation, the urine was subjected to a one-step cleanup on a commercially available immunoaffinity chromatography cartridge. The analytes were detected by liquid chromatography-negative-ion electrospray tandem mass spectrometry using deuterium-labelled internal standards. The method was validated as a quantitative confirmatory method according to European Commission Decision 2002/657/EC. The evaluated parameters were: linearity, specificity, precision (repeatability and intra-laboratory reproducibility), recovery, decision limit, detection capability and ruggedness. The decision limits (CCalpha) obtained, were between 0.56 and 0.68 microgL(-1); recovery above 66% for all the analytes. Repeatability was between 1.4 and 5.3% and within-laboratory reproducibility between 1.9 and 16.1% for the six resorcylic acid lactones.
Dusi
Guglielmo
G
Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, "B. Ubertini", Via Bianchi 7, 25124 Brescia, Italy.
Bozzoni
Eros
E
Assini
Walter
W
Tognoli
Nadia
N
Gasparini
Mara
M
Ferretti
Enrica
E
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004968:Estrogens, Non-Steroidal / Q000302:isolation & purification / Q000652:urine*; D015203:Reproducibility of Results; D015813:Substance Abuse Detection / Q000379:methods*; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000379:methods*; D015025:Zearalenone / Q000302:isolation & purification / Q000652:urine; D015029:Zeranol / Q000302:isolation & purification / Q000652:urine*
D004968:Estrogens, Non-Steroidal; D015025:Zearalenone; D015029:Zeranol; C028226:taleranol