Skip to main content
SpringerLink
Log in

Preparation of Monolithic Imprinted Stationary Phase for Clenbuterol by In Situ Polymerization and Application in Biological Samples Pretreatment

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Re G, Badino P, Novelli A, Girardi C (1997) Effects of clenbuterol as a repartitioning agent on [beta]-adrenoceptor concentrations in heart, bronchi and brain of veal calves. Vet J 153(1):63–70. doi:10.1016/s1090-0233(97)80009-3

    Article  CAS  Google Scholar 

  2. Kearns CF, McKeever KH (2009) Clenbuterol and the horse revisited. Vet J 182(3):384–391. doi:10.1016/j.tvjl.2008.08.021

    Article  CAS  Google Scholar 

  3. Waterfield CJ, Jairath M, Asker DS, Timbrell JA (1995) The biochemical effects of clenbuterol: with particular reference to taurine and muscle damage. Eur J Pharmacol Environ Toxicol Pharmacol Section 293(2):141–149. doi:10.1016/0926-6917(95)00010-0

    Article  CAS  Google Scholar 

  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

    Article  CAS  Google Scholar 

  5. Masci G, Casati G, Crescenzi V (2001) Synthesis and LC characterization of clenbuterol molecularly imprinted polymers. J Pharm Biomed Anal 25(2):211–217. doi:10.1016/s0731-7085(00)00477-5

    Article  CAS  Google Scholar 

  6. Vlatakis G, Andersson LI, Muller R, Mosbach K (1993) Drug assay using antibody mimics made by molecular imprinting. Nature 361(6413):645–647

    Article  CAS  Google Scholar 

  7. Haginaka J (2005) Selectivity of affinity media in solid-phase extraction of analytes. Trends Anal Chem 24(5):407–415

    Article  CAS  Google 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

    Article  CAS  Google 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

    Article  CAS  Google Scholar 

  10. Brambilla G, Fiori M, Rizzo B, Crescenzi V, Masci G (2001) Use of molecularly imprinted polymers in the solid-phase extraction of clenbuterol from animal feeds and biological matrices. J Chromatogr B 759(1):27–32. doi:10.1016/s0378-4347(01)00199-2

    Article  CAS  Google Scholar 

  11. Andrea P, Miroslav S, Silvia S, Stanislav M (2001) A solid binding matrix/molecularly imprinted polymer-based sensor system for the determination of clenbuterol in bovine liver using differential-pulse voltammetry. Sens Actuator B 76(1–3):286–294

    Article  Google 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

    Article  CAS  Google 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

    Article  CAS  Google Scholar 

  14. Yin J, Yang G, Chen Y (2005) Rapid and efficient chiral separation of nateglinide and its l-enantiomer on monolithic molecularly imprinted polymers. J Chromatogr A 1090(1–2):68–75. doi:10.1016/j.chroma.2005.06.078

    Article  CAS  Google Scholar 

  15. Amut E, Fu Q, Fang Q, Liu R, Xiao AP, Zeng AG, Chang C (2010) In situ polymerization preparation of chiral molecular imprinting polymers monolithic column for amlodipine and its recognition properties study. J Polym Res 17(3):401–409. doi:10.1007/s10965-009-9326-3

    Article  CAS  Google Scholar 

  16. Rao TP, Kala R, Daniel S (2006) Metal ion-imprinted polymers: novel materials for selective recognition of inorganics. Anal Chim Acta 578(2):105–116. doi:10.1016/j.aca.2006.06.065

    Article  CAS  Google Scholar 

  17. Xu ZF, Kuang DZ, Feng YL, Zhang FX (2010) Combination of hydrophobic effect and electrostatic interaction in imprinting for achieving efficient recognition in aqueous media. Carbohydr Polym 79(3):642–647. doi:10.1016/j.carbpol.2009.09.010

    Article  CAS  Google Scholar 

  18. Zhu Q-Z, Haupt K, Knopp D, Niessner R (2002) Molecularly imprinted polymer for metsulfuron-methyl and its binding characteristics for sulfonylurea herbicides. Anal Chim Acta 468(2):217–227. doi:10.1016/s0003-2670(01)01437-4

    Article  CAS  Google Scholar 

  19. Haginaka J, Takehira H, Hosoya K, Tanaka N (1999) Uniform-sized molecularly imprinted polymer for (S)-naproxen selectively modified with hydrophilic external layer. J Chromatogr A 849(2):331–339. doi:10.1016/s0021-9673(99)00570-1

    Article  CAS  Google Scholar 

  20. Xiang Y, Chen D (2007) Preparation of a novel pH-responsive silver nanoparticle/poly(HEMA-PEGMA-MAA) composite hydrogel. Eur Polym J 43(10):4178–4187. doi:10.1016/j.eurpolymj.2007.08.005

    Article  CAS  Google Scholar 

  21. Verliefde ARD, Cornelissen ER, Heijman SGJ, Verberk J, Amy GL, Van der Bruggen B, van Dijk JC (2008) The role of electrostatic interactions on the rejection of organic solutes in aqueous solutions with nanofiltration. J Memb Sci 322(1):52–66. doi:10.1016/j.memsci.2008.05.022

    Article  CAS  Google Scholar 

  22. Herraez-Hernandez R, Campins-Falco P (2000) Derivatization of ephedrine with o-phthaldialdehyde for liquid chromatography after treatment with sodium hypochlorite. J Chromatogr A 893(1):69–80

    Article  CAS  Google Scholar 

  23. Vamvaca K, Jelesarov I, Hilvert D (2008) Kinetics and thermodynamics of ligand binding to a molten globular enzyme and its native counterpart. J Mol Biol 382(4):971–977. doi:10.1016/j.jmb.2008.07.049

    Article  CAS  Google Scholar 

  24. van Zoelen EJJ, Kramer RH, van Moerkerk HTB, Veerkamp JH (1998) The use of nonhomologous scatchard analysis in the evaluation of ligand–protein interactions. Trends Pharmacol Sci 19(12):487–490

    Article  Google Scholar 

Download references

Acknowledgments

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.

Author information

Authors and Affiliations

  1. Faculty of Pharmacy, School of Medicine, Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China

    Yifen Luo, Ping Huang, Qiang Fu, Wei Du, Sijuan Sun, Ya Li, Meng Liu & Chun Chang

Authors
  1. Yifen Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ping Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qiang Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wei Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sijuan Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ya Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Meng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Chun Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Qiang Fu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Luo, Y., Huang, P., Fu, Q. et al. Preparation of Monolithic Imprinted Stationary Phase for Clenbuterol by In Situ Polymerization and Application in Biological Samples Pretreatment. Chromatographia 74, 693–701 (2011). https://doi.org/10.1007/s10337-011-2129-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-011-2129-9

Keywords

Search

Navigation