Proceeding of Veterinary and Animal Science Days 2016, 8th- 10th June, Milan, Italy 

HAF © 2013 

Vol. III, No. 1s  ISSN: 2283-3927 

   

 

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Keywords 

Bovine, Teeth, β2-agonists, 

Residues, HPLC-MS/MS. 
 

CORRESPONDING AUTHOR 

Maria Nobile 

maria.nobile1@unimi.it 

 

JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

Detection of seven β2-agonists in teeth 

by LC-MS/MS: preliminary results. 

M. Nobilea*, S. Panseria, F. Ariolia, L. M. Chiesaa 

 

 

 

 a Department of Health, Animal Science and Food Safety, University of 
Milan, Via Celoria 10, 20133 Milan, Italy 

 

 

 

Abstract 
β2-agonists are powerful tocolytic (the only use permitted in cattle) and bronchodilator agents, but 

may also be administered as growth promoters to improve the production of lean meat increasing also 

the lipolytic activity. Although the European Union (EU), China and other Asian countries have banned 

the use of β2-agonists for growth promoting purposes (European Union, 2003), the United States of 

America (USA) authorised ractopamine as a feed additive for swine, cattle and turkey. These veterinary 

drugs, generally show high clearance rates in the conventional biological matrices, as well as urine, liver 

and muscle, making difficult their detection (Wu, 2014). For this reason, we suggested bovine teeth as 

a new unconventional matrix of accumulation in a more long-time window, for the detection of 

cimaterol, clenbuterol, isoxsuprine, mabuterol, ractopamine, salbutamol and terbutaline. In literature, 

the few studies on teeth are limited to human (Andra, 2015) and are absent for veterinary medicine. 

The samples extracted by a simple liquid extraction step with ethyl acetate:tert-butyl methyl ether (4:1, 

v/v)  after washing and pulverization of teeth, through a ball mill, were analysed using a liquid 

chromatography tandem mass spectrometry (LC-MS/MS) confirmatory method validated according to 

the Commission Decision 2002/657/EC criteria (European Union, 2002). Teeth from 8 veal calves, 

administered per os with 80 mg day-1 oral ractopamine for 32 days, and from seven random bovines 

from the food chain were collected at the slaughterhouse to test the suitability of this matrix. The 

results demonstrated ractopamine presence in teeth from the treated animals (average concentration 

8.90 ng g-1). Isoxsuprine was found in a control sample (13.67 ng g-1), demonstrating the effectiveness 

of this matrix as a powerful tool to ensure illegal treatment.  
 

References 

Andra, S.S., Austin, C., Arora, M., 2015. Tooth matrix analysis for biomonitoring of organic chemical exposure: Current status, challenges, 

and opportunities. Environ. Res. 142, 387-406. 

European Union, 2002. Commission Decision concerning the performance of analytical methods and the interpretation of results. Off. J. Eur. 

Comm. L221, 8-36. 

European Union, 2003. Council directive 2003/74/EC amending Council Directive 96/22/EC concerning the prohibition on the use of stock 

farming of certain substances having a hormonal or thyrostatic action and of β-agonists. Off. J. Eur. Union. L262, p.17. 

Wu, J., Liu, X., Peng, Y., 2014. Determination of ractopamine in pig hair using liquid chromatography with tandem mass spectrometric 

detection. J. Pharmacol. Toxicol. Meth. 69, 211-216. 

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