Criteria to distinguish between natural situations and illegal use of boldenone, boldenone esters and boldione in cattle 2. Direct measurement of 17beta-boldenone sulpho-conjugate in calf urine by liquid chromatography--high resolution and tandem mass spectrometry.
Boldenone is banned in the European Union (Directive 96/22/EC) as growth promoter for meat producing animals. Boldione (ADD), boldenone and boldenone esters (mainly the undecylenate form) are commercially available as anabolic preparations, either to the destination of human, horse or cattle. Since the late 90s, the natural occurrence of boldenone metabolites has been reported in cattle. According to EU regulation, the unambiguous demonstration of boldenone administration in bovine urine should be provided on the basis of boldenone identification in the corresponding conjugate fraction. An analytical method has been developed and validated according to current standards with main concern to the measurement of intact 17beta-boldenone-sulphate. The analytical procedure included direct extraction-purification of target analyte on octadecylsilyl cartridges and direct detection of phase II metabolite by liquid chromatography (negative electrospray), tandem mass spectrometry (QqQ) or high resolution mass spectrometry (Orbitrap). Decision limit (CCalpha) and detection capability (CCbeta) were respectively 0.2 microg L(-1) and 0.4 microg L(-1) on triple quadrupole and 0.1 microg L(-1) and 0.2 microg L(-1) on hybrid system. The method was successfully applied to the analysis of incurred samples collected in different experiments. 17beta-Boldenone-sulphate was measurable up to 36h after oral administration of boldione, and 30 days after 17beta-boldenone undecylenate intra-muscular injection. This conjugate form was never detected in non-treated animals, confirming its status of definitive candidate marker for boldenone administration in calf.
Destrez
Blandine
B
Laboratoire d'Etude des Résidus et Contaminants dans les Aliments (LABERCA), Ecole Nationale Vétérinaire de Nantes (ENVN), BP 50707, Route de Gachet, 44307 Nantes Cedex 3, France.
Bichon
Emmanuelle
E
Rambaud
Lauriane
L
Courant
Frédérique
F
Monteau
Fabrice
F
Pinel
Gaud
G
Antignac
Jean-Philippe
JP
Le Bizec
Bruno
B
D045930:Anabolic Agents / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics / Q000652:urine*; D000730:Androstadienes / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics / Q000652:urine*; D000821:Animal Feed; D000818:Animals; D002417:Cattle / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D004952:Esters / Q000737:chemistry*; D020719:Glucuronides / Q000378:metabolism; D006736:Horses; D006801:Humans; D008297:Male; D008657:Metabolic Clearance Rate; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D015813:Substance Abuse Detection; D013431:Sulfates / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism / Q000652:urine*; D053719:Tandem Mass Spectrometry; D013739:Testosterone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics / Q000652:urine
C545457:17-boldenone-sulfate; D045930:Anabolic Agents; D000730:Androstadienes; D004952:Esters; D020719:Glucuronides; D013431:Sulfates; C006573:1,4-androstadiene-3,17-dione; D013739:Testosterone; C013915:boldenone