A method for the determination of five β2-agonists in pig feed, liver, meat, and urine samples by micellar electrokinetic chromatography was developed. The separation and detection conditions were optimized in terms of SDS concentration, running buffer pH and concentration, capillary temperature. Under the optimized conditions (15 mmol/L borate, pH=10, 15 mmol/L SDS, and 25 ℃), all five analytes in the spiking samples were separated and identified within 18 min. Good linearity (R2>0.999) was obtained for the analytes in the range of 0.1-40 μg/mL with the limit of detection (S/N ≥ 3) in the range of 0.05-0.08 μg/mL. The satisfactory recoveries of cimaterol, ractopamine and clenbuterol were obtained in the range of 93.1%-95.8%, whereas the recoveries of salbutamol and terbutaline were at 52.3% and 69.4%, respectively. The relative standard deviations of peak area and migration time of the analytes were less than 4.86% and 1.25%, respectively. The method was successfully applied to monitor the presence of the selected β2-agonists in pig feed, liver, meat and urine samples.
[1] Stoffel B, Meyer H H D. Effects of the beta-adrenergic agonist clenbuterol in cows: lipid metabolism, milk production, pharmacokinetics, and residues [J]. Journal of Animal Science, 1993, 71 (7):1 875-1 881.
[2] Mersmann H J. Overview of the effects of beta-adrenergic receptor agonists on animal growth including mechanisms of action [J].Journal of Animal Science, 1998, 76 (1): 160-172.
[3] Mazzanti G, Daniele C, Boatto G, et al. New beta-adrenergic agonists used illicitly as growth promoters in animal breeding: chemical and pharmacodynamic studies [J]. Toxicology, 2003, 187(2/3): 91-99.
[4] Brambilla G, Cenci T, Franconi F, et al. Clinical and pharmacological profile in a clenbuterol epidemic poisoning of contaminated beef meat in Italy [J]. Toxicology Letters, 2000, 114 (1-3): 47-53.
[5] Kuiper H A, Noordam M Y, van Dooren-Flipsen M M H, et al. Illegal use of beta-adrenergic agonists: European community [J]. Journal of Animal Science, 1998, 76 (1): 195-207.
[6] Chen Q, Fan L Y, Zhang W, et al. Separation and determination of abused drugs clenbuterol and salbutamol from complex extractants in swine feed by capillary zone electrophoresis with simple pretreatment [J]. Talanta, 2008, 76 (2): 282-287.
[7] Pleadin J, Vulié A, Mitak M, et al. Determination of clenbuterol residues in retinal tissue of food-producing pigs [J]. Journal of Analytical Toxicology, 2011, 35 (1):28-31.
[8] Zhang H C, Liu G Y, Chai C Y. A novel amperometric sensor based on screen-printed electrode modified with multi-walled carbon nanotubes and molecularly imprinted membrane for rapid determination of ractopamine in pig urine [J].Sensors and Actuators B: Chemical, 2012, 168: 103-110.
[9] He P L, Shen L, Liu R Y, et al. Direct detection of beta-agonists by use of gold nanoparticle-based colorimetric assays [J].Analytical Chemistry, 2011, 83 (18):6 988-6 995.
[10] Lin Q B, Zhao X T, Song H, et al. Immunoaffinity chromatography purification and ultra-high-performance liquid chromatography-tandem mass spectrometry determination of four beta-agonists in beef [J]. Food Additives & Contaminants: Part A, 2012, 29 (6): 935-941.
[11] Liu C, Ling W T, Xu W D, et al. Simultaneous determination of 20 beta-agonists in pig muscle and liver by high-performance liquid chromatography/tandem mass spectrometry [J]. Journal of AOAC International, 2011, 94 (2): 420-427.
[12] Wang X J, Zhang F, Ding F, et al. Simultaneous determination of 12 beta-agonists in feeds by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry [J]. Journal of Chromatography A, 2013, 1278:82-88.
[13] Caban M, Stepnowski P, Kwiatkowski M, et al. Determination of beta-blockers and beta-agonists using gas chromatography and gas chromatography-mass spectrometry-A comparative study of the derivatization step [J]. Journal of Chromatography A, 2011, 1218 (44): 8 110-8 122.
[14] Di Corcia D, Morra V, Pazzi M, et al. Simultaneous determination of beta(2)-agonists in human urine by fast-gas chromatography/mass spectrometry: method validation and clinical application [J]. Biomedical Chromatography, 2010, 24(4): 358-366.
[15] Liu W, Zhang L, Wei Z Y, et al. Analysis of beta-agonists and beta-blockers in urine using hollow fibre-protected liquid-phase microextraction with in situ derivatization followed by gas chromatography/mass spectrometry [J]. Journal of Chromatography A,2009, 1216 (28): 5 340-5 346.
[16] He L M, Su Y J, Zeng Z L, et al. Determination of ractopamine and clenbuterol in feeds by gas chromatography-mass spectrometry [J]. Animal Feed Science and Technology, 2007, 132 (3/4): 316-323.
[17] Ramos F, Cristino A, Carrola P, et al. Clenbuterol food poisoning diagnosis by gas chromatography-mass spectrometric serum analysis [J]. Analytica Chimica Acta, 2003, 483 (1-2): 207-213.
[18] Wang P L, Li Y, Wang R G, et al. Determination of beta(2)-agonists in porcine urine by molecularly imprinted solid phase extraction followed ultra-performance liquid chromatography-tandem mass spectrometry detection [J]. Analytical Letters,2013, 46 (5):734-744.
[19] Shi Y F, Huang Y, Duan J P, et al. Field-amplified on-line sample stacking for separation and determination of cimaterol, clenbuterol and salbutamol using capillary electrophoresis [J]. Journal of Chromatography A, 2006, 1125 (1):124-128.
[20] Zhou S S, Wang Y Q, De Beer T, et al. Simultaneous separation of eight β-adrenergic drugs using titanium dioxide nanoparticles as additive in capillary electrophoresis [J]. Electrophoresis, 2008,29 (11): 2 321-2 329.
[21] Anurukvorakun O, Buchberger W, Himmelsbach M, et al. A sensitive non-aqueous capillary electrophoresis-mass spectrometric method for multiresidue analyses of beta-agonists in pork [J]. Biomedical Chromatography, 2010, 24 (6): 588-599.
[22] Wang W Y, Zhang Y L, Wang J Y, et al. Determination of beta-agonists in pig feed, pig urine and pig liver using capillary electrophoresis with electrochemical detection [J]. Meat science, 2010,85 (2):302-305.
[23] Bao Y, Yang F, Yang X R. Capillary electrophoresis coupled with electrochemiluminescence for the facile separation and determination of salbutamol and clenbuterol in urine [J]. Electroanalysis, 2012, 24 (7): 1 597-1 603.
[24] Mazzarino M, de la Torre X, Mazzei F, et al. Rapid screening of beta-adrenergic agents and related compounds in human urine for anti-doping purpose using capillary electrophoresis with dynamic coating [J]. Journal of Separation Science, 2009, 32 (20): 3 562-3 570.
[25] Pappas T J, Gayton-Ely M, Holland L A. Recent advances in micellar electrokinetic chromatography [J]. Electrophoresis, 2005, 26 (4/5): 719-734.
[26] Fritz J S. The role of organic solvents in the separation of nonionic compounds by capillary electrophoresis [J]. Electrophoresis, 2003, 24 (10): 1 530-1 536.
[27] Ding Y S, Qi Y L, Suo X M. Rapid determination of beta(2)-agonists in urine samples by microchip micellar electrokinetic chromatography with pulsed electrochemical detection [J]. Analytical Methods, 2013, 5 (10):2 623-2 629.
[28] Tanford C. Micelle shape and size [J]. The Journal of Physical Chemistry,1972, 76 (21): 3 020-3 024.
