Preparation of Monolithic Imprinted Stationary Phase for Clenbuterol by In Situ Polymerization and Application in Biological Samples Pretreatment
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A clenbuterol (CLB) molecule-imprinted monolithic stationary phase (MIMSP) with specific recognition for CLB and some other β2-adrenergic receptor agonists was prepared by in situ polymerization technique utilizing methacrylic acid as a functional monomer, ethylene glycol dimethacrylate (EDMA) as a cross-linking reagent, and low polar solvents (toluene and dodecanol) as porogenic solvents. The optimal polymerization conditions were as follows: the molar ratio of template:monomer:initiator was 5:20:1, EDMA was 85% (v/v) in the total volume of monomer and EDMA, and toluene was 18% (v/v) in the total mixed porogen. The selectivity of the stationary phase for CLB and other β2-adrenergic receptor agonists was evaluated by high performance liquid chromatography. Scatchard analysis was employed to explore the recognition mechanism. Then the CLB-MIMSP was used as a solid phase extraction (SPE) stationary phase for concentration and purification of CLB from pig liver samples. The results showed that the obtained CLB-MIMSP possessed high selectivity towards CLB and moderate selectivity towards some other β2-adrenergic receptor agonists with characteristics of easy-made. The limit of detection was 10 ng g−1, and recoveries of CLB were 99.16–113.06% with RSD 4.55–11.81% for the spiked pig liver samples. The CLB-MIMSP could be a promising SPE absorbent in CLB biological sample pretreatment.
KeywordsColumn liquid chromatography Clenbuterol Molecule-imprinted monolithic stationary phase In situ polymerization Solid phase extraction Biological samples pretreatment
Financial support of this work by National Natural Science Foundations of China (No.30873193) to Qiang FU is gratefully acknowledged. The authors also thank professor Jun Haginaka from Mukogawa Women’s University, Japan, for his helpful discussion in molecularly imprinted polymers preparation.
- 4.Mazzanti G, Di Sotto A, Daniele C, Battinelli L, Brambilla G, Fiori M, Loizzo S, Loizzo A (2007) A pharmacodynamic study on clenbuterol-induced toxicity: [beta]1- and [beta]2-adrenoceptors involvement in guinea-pig tachycardia in an in vitro model. Food Chem Toxicol 45(9):1694–1699. doi: 10.1016/j.fct.2007.03.002 CrossRefGoogle Scholar
- 8.Sambe H, Hoshina K, Haginaka J (2007) Molecularly imprinted polymers for triazine herbicides prepared by multi-step swelling and polymerization method: their application to the determination of methylthiotriazine herbicides in river water. J Chromatogr A 1152(1–2):130–137. doi: 10.1016/j.chroma.2006.09.003 CrossRefGoogle Scholar
- 9.Zhang QQ, Fu Q, Amut EJ, Fang Q, Zeng AG, Chang C (2009) Preparation and evaluation of propranolol-imprinted monolithic stationary phase by in situ technique and application in analysis of propranolol in biological samples. Anal Lett 42(3):536–554. doi: 10.1080/00032710802677084 CrossRefGoogle Scholar
- 12.Bruins CHP, Jeronimus-Stratingh CM, Ensing K, van Dongen WD, de Jong GJ (1999) On-line coupling of solid-phase extraction with mass spectrometry for the analysis of biological samples: I. determination of clenbuterol in urine. J Chromatogr A 863(1):115–122. doi: 10.1016/s0021-9673(99)00959-0 CrossRefGoogle Scholar
- 13.Ou JJ, Kong L, Pan CS, Su XY, Lei XY, Zou HF (2006) Determination of dl-tetrahydropalmatine in Corydalis yanhusuo by l-tetrahydropalmatine imprinted monolithic column coupling with reversed-phase high performance liquid chromatography. J Chromatogr A 1117(2):163–169. doi: 10.1016/j.chroma.2006.03.084 CrossRefGoogle Scholar