Analytical and Bioanalytical Chemistry

, Volume 407, Issue 6, pp 1745–1752 | Cite as

Molecularly imprinted microspheres synthesized by a simple, fast, and universal suspension polymerization for selective extraction of the topical anesthetic benzocaine in human serum and fish tissues

  • Hui Sun
  • Jia-Ping LaiEmail author
  • Fang Chen
  • De-Rong Zhu
Research Paper


A simple, fast, and universal suspension polymerization method was used to synthesize the molecularly imprinted microspheres (MIMs) for the topical anesthetic benzocaine (BZC). The desired diameter (10–20 μm) and uniform morphology of the MIMs were obtained easily by changing one or more of the synthesis conditions, including type and amount of surfactant, stirring rate, and ratio of organic to water phase. The MIMs obtained were used as a molecular-imprinting solid-phase-extraction (MISPE) material for extraction of BZC in human serum and fish tissues. The MISPE results revealed that the BZC in these biosamples could be enriched effectively after the MISPE operation. The recoveries of BZC on MIMs cartridges were higher than 90 % (n = 3). Finally, an MISPE-HPLC method with UV detection was developed for highly selective extraction and fast detection of trace BZC in human serum and fish tissues. The developed method could also be used for the enrichment and detection of BZC in other complex biosamples.


Molecularly imprinted microspheres Benzocaine Aqueous-suspension polymerization Solid-phase extraction Human serum Fish tissues 



This work was supported by the National Natural Science Foundation of China (21107018, 21477026), Science and Technology Key Project of Ministry of Education (212129), Natural Science Foundation of Guangdong Province (S2012010008172).

Supplementary material

216_2014_8420_MOESM1_ESM.pdf (450 kb)
ESM 1 (PDF 317 kb)


  1. 1.
    De Araujo DR, Padula C, Cereda CMS, Tofoli GR, Brito RB, de Paula E, Nicoli S, Santi P (2010) Bioadhesive films containing benzocaine: correlation between in vitro permeation and in vivo local anesthetic effect. Pharm Res 27:1677–1686Google Scholar
  2. 2.
    De Melo NFS, Grillo R, Guilherme VA, de Araujo DR, de Paula E, Rosa AH, Fraceto LF (2011) Poly(lactide-co-glycolide) nanocapsules containing benzocaine: influence of the composition of the oily nucleus on physico-chemical properties and anesthetic activity. Pharm Res 28:1984–1994Google Scholar
  3. 3.
    Brock WJ, Bell TA (2012) The in vitro and in vivo genotoxicity of benzocaine: a brief communication. Int J Toxicol 31:222–227Google Scholar
  4. 4.
    Novaro GM, Aronow HD, Militello MA, Garcia MJ, Sabik EM (2003) Benzocaine-induced methemoglobinemia: experience from a high-volume transesophageal echocardiography laboratory. J Am Soc Echocardiogr 16:170–175Google Scholar
  5. 5.
    Moore TJ, Walsh CS, Cohen MR (2004) Reported adverse event cases of methemoglobinemia associated with benzocaine products. Arch Int Med 164:1192–1196Google Scholar
  6. 6.
    Kane GC, Hoehn SM, Behrenbeck TR, Mulvagh SL (2007) Benzocaine-induced methemoglobinemia based on the Mayo Clinic experience from 28 478 transesophageal echocardiograms - incidence, outcomes, and predisposing factors. Arch Int Med 167:1977–1982Google Scholar
  7. 7.
    Srikanth MS, Kahlstrom R, Fox SR, Fox ER, Fox KM (2005) Topical benzocaine (Hurricaine((R))) induced methemoglobinemia during endoscopic procedures in gastric bypass patients. Obes Surg 15:584–590Google Scholar
  8. 8.
    Taleb M, Ashraf Z, Valavoor S, Tinkel J (2013) Evaluation and management of acquired Methemoglobinemia associated with topical benzocaine use. Am J Cardiol Drugs 13:325–330Google Scholar
  9. 9.
    Meinertz JR, Stehly GR, Hubert TD, Bernardy JA (1999) Liquid chromatographic determination of benzocaine and N-acetylbenzocaine in the edible fillet tissue from rainbow trout. J Chromatogr A 855:255–260Google Scholar
  10. 10.
    Perez-Lozano P, Garcia-Montoya E, Orriols A, Minarro M, Tico JR, Sune-Negre JM (2005) A new validated method for the simultaneous determination of benzocaine, propylparaben and benzyl alcohol in a bioadhesive gel by HPLC. J Pharm Biomed Anal 39:920–927Google Scholar
  11. 11.
    Dejmkova H, Vokalova V, Zima J, Barek J (2011) Electroanalysis 23:662–666Google Scholar
  12. 12.
    Scherpenisse P, Bergwerff AA (2007) Determination of residues of tricaine in fish using liquid chromatography tandem mass spectrometry. Anal Chim Acta 586:407–410Google Scholar
  13. 13.
    De Orsi D, Pellegrini M, Marchei E, Nebuloni P, Gallinella B, Scaravelli G, Martufi A, Gagliardi L, Pichini S (2009) High performance liquid chromatography-diode array and electrospray-mass spectrometry analysis of vardenafil, sildenafil, tadalafil, testosterone and local anesthetics in cosmetic creams sold on the internet web sites. J Pharm Biomed Anal 50:362–369Google Scholar
  14. 14.
    Kachoosangi RT, Wildgoose GG, Compton RG (2008) Using capsaicin modified multiwalled carbon nanotube based electrodes and p-chloranil modified carbon paste electrodes for the determination of amines: application to benzocaine and lidocaine. Electroanalysis 20:2495–2500Google Scholar
  15. 15.
    Zhang XR, Baeyens WRG, Vanderweken G, Calokerinos AC, Imai K (1995) Chemiluminescence determination of some local-anesthetics. Anal Chim Acta 303:137–142Google Scholar
  16. 16.
    Pasekova H, Polasek M (2000) Determination of procaine, benzocaine and tetracaine by sequential injection analysis with pel-manganate-induced chemiluminescence detection. Talanta 52:67–75Google Scholar
  17. 17.
    Porra R, Berri S, Gagliardi L, Chimenti P, Granese A, De Orsi D, Carpani I, Tonelli D (2004) Development of an HPLC method for the identification and dosage of non-allowed substances in cosmetic products. Part I: local anaesthetics and antihistaminics. Anal Bioanal Chem 380:767–772Google Scholar
  18. 18.
    Nochetto CB, Reimschuessel R, Gieseker C, Cheely CS, Carson MC (2009) Determination of tricaine residues in fish by liquid chromatography. J AOAC Int 92:1241–1247Google Scholar
  19. 19.
    Duan ZJ, Fan LP, Fang GZ, Yi JH, Wang S (2011) Novel surface molecularly imprinted sol-gel polymer applied to the online solid phase extraction of methyl-3-quinoxaline-2-carboxylic acid and quinoxaline-2-carboxylic acid from pork muscle. Anal Bioanal Chem 401:2291–2299Google Scholar
  20. 20.
    Baldim IM, Souza MCD, Souza JCJD, Figueiredo EC, Martins I (2012) Application of the molecularly imprinted solid-phase extraction to the organophosphate residues determination in strawberries. Anal Bioanal Chem 404:1959–1966Google Scholar
  21. 21.
    You QP, Peng MJ, Zhang YP, Guo JF, Shi SY (2014) Preparation of magnetic dummy molecularly imprinted polymers for selective extraction and analysis of salicylic acid in Actinidia chinensis. Anal Bioanal Chem 406:831–839Google Scholar
  22. 22.
    Su Z, Zhai HY, Chen ZG, Zhou Q, Li JM, Liu ZP (2014) Molecularly imprinted solid-phase extraction monolithic capillary column for selective extraction and sensitive determination of safranine T in wolfberry. Anal Bioanal Chem 406:1551–1556Google Scholar
  23. 23.
    Moraes GDI, da Silva LMR, dos Santos-Neto AJ, Florenzano FH, Figueiredo EC (2013) A new restricted access molecularly imprinted polymer capped with albumin for direct extraction of drugs from biological matrices: the case of chlorpromazine in human plasma. Anal Bioanal Chem 405:7687–7696Google Scholar
  24. 24.
    Lai JP, Xie L, Sun H, Chen F (2013) Synthesis and evaluation of molecularly imprinted polymeric microspheres for highly selective extraction of an anti-AIDS drug emtricitabine. Anal Bioanal Chem 405:4269–4275Google Scholar
  25. 25.
    Lai JP, Niessner R, Knopp D (2004) Benzo[a]pyrene imprinted polymers: synthesis, characterization and SPE application in water and coffee samples. Anal Chim Acta 522:137–144Google Scholar
  26. 26.
    Lai JP, Yang ML, Niessner R, Knopp D (2007) Molecularly imprinted microspheres and nanospheres for di(2-ethylhexyl)phthalate prepared by precipitation polymerization. Anal Bioanal Chem 389:405–412Google Scholar
  27. 27.
    Pan GQ, Zhang Y, Ma Y, Li CX, Zhang HQ (2011) Efficient one-pot synthesis of water-compatible molecularly imprinted polymer microspheres by facile RAFT precipitation polymerization. Angew Chem Int Ed 50:11731–11734Google Scholar
  28. 28.
    Lai JP, Lu XY, Lu CY, Ju HF, He XW (2001) Preparation and evaluation of molecularly imprinted polymeric microspheres by aqueous suspension polymerization for use as a high-performance liquid chromatography stationary phase. Anal Chim Acta 442:105–111Google Scholar
  29. 29.
    Lenain P, Di Mavungu JD, Dubruel P, Robbens J, De Saeger S (2012) Development of suspension polymerized molecularly imprinted beads with metergoline as template and application in a solid-phase extraction procedure toward ergot alkaloids. Anal Chem 84:10411–10418Google Scholar
  30. 30.
    Geng LY, Kou X, Lei JD, Su HJ, Ma GH, Su ZG (2012) Preparation, characterization and adsorption performance of molecularly imprinted microspheres for erythromycin using suspension polymerization. J Chem Technol Biotechnol 87:635–642Google Scholar
  31. 31.
    Haginaka J, Tabo H, Matsunaga H (2012) Preparation of molecularly imprinted polymers for organophosphates and their application to the recognition of organophosphorus compounds and phosphopeptides. Anal Chim Acta 748:1–8Google Scholar
  32. 32.
    Yang YK, Fang GZ, Wang XM, Pan MF, Qian HL, Liu HL, Wang S (2014) Sensitive and selective electrochemical determination of quinoxaline-2-carboxylic acid based on bilayer of novel poly(pyrrole) functional composite using one-step electro-polymerization and molecularly imprinted poly(o-phenylenediamine). Anal Chim Acta 806:136–143Google Scholar
  33. 33.
    Xu SY, Guo CJ, Li YX, Yu ZR, Wei CH, Tang YW (2014) Methyl parathion imprinted polymer nanoshell coated on the magnetic nanocore for selective recognition and fast adsorption and separation in soils. J Hazard Mater 264:34–41Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.College of Environmental Science & EngineeringGuangzhou UniversityGuangzhouChina
  2. 2.School of Chemistry & EnvironmentSouth China Normal UniversityGuangzhouChina
  3. 3.School of PharmacyGuangdong Medical CollegeDongwanChina

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