Purification of clenbuterol-like beta2-agonist drugs of new generation from bovine urine and hair by alpha1-acid glycoprotein affinity chromatography and determination by gas chromatography-mass spectrometry.
The control of illegal use of clenbuterol and other beta(2)-agonist drugs as growth promoters in the European Union countries has led to outlaw practices for synthesizing new concept molecules, showing similar biological activity but not detectable by test methods usually employed to perform the official monitoring programmes. The synthesis schemes of some beta(2)-agonist compounds, formally derived from clenbuterol, were found out by Italian detective authorities. These compounds were synthesised ex novo in our laboratories: then, both their molecular structures and biological activities were characterised. In this paper, we describe different strategies for purifying some beta(2)-agonist drugs of new concept, more hydrophobic than clenbuterol. A two-step clean up procedure, prior to gas chromatography-mass spectrometry analysis, was developed for the multi-residue determination of these beta(2)-agonists from bovine hair and urine. The purification strategy we chose was based on adsorption solid phase extraction and, subsequently, on specific molecular recognition by affinity chromatography. The affinity columns were homemade by coupling bovine alpha(1)-acid glycoprotein, a plasmatic acceptor for basic drugs, to a chromatographic support; their effectiveness for purifying new beta(2)-agonists was discussed. The data about method recoveries and repeatability were also reported.
Gallo
Pasquale
P
Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute 2, 80055 Portici, NA, Italy. p.gallo@izsmportici.it <p.gallo@izsmportici.it>
Brambilla
Gianfranco
G
Neri
Bruno
B
Fiori
Maurizio
M
Testa
Cecilia
C
Serpe
Luigi
L
D000318:Adrenergic beta-Agonists / Q000302:isolation & purification* / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002845:Chromatography / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002976:Clenbuterol / Q000031:analogs & derivatives* / Q000302:isolation & purification* / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006197:Hair / Q000378:metabolism; D007202:Indicators and Reagents / Q000494:pharmacology; D009961:Orosomucoid / Q000737:chemistry; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods; D015813:Substance Abuse Detection / Q000379:methods*; D014556:Urine / Q000737:chemistry
D000318:Adrenergic beta-Agonists; D007202:Indicators and Reagents; D009961:Orosomucoid; D002976:Clenbuterol