Preparation and characterization of a molecularly imprinted polymer by grafting on silica supports: a selective sorbent for patulin toxin

  • Dayun ZhaoEmail author
  • Jingfu Jia
  • Xuelei Yu
  • Xiangjun Sun
Original Paper


A new molecularly imprinted polymer (MIP) has been prepared on silica beads using the radical “grafting from” polymerization method for selective extraction of minor contaminant mycotoxin of patulin (PTL). After the introduction of amino groups onto the silica surface with 3-aminopropyltriethoxysilane, azo initiator onto the silica surface was achieved by the reaction of surface amino groups with 4,4′-azobis(4-cyanopentanoic acid). The scale-up synthesis of MIP was then carried out in the presence of 6-hydroxynicotinic acid as template substitute, functional, and cross-linking monomers. The prepared sorbent was characterized using FT-IR spectroscopy, scanning electron microscopy, elemental analysis, and the adsorption–desorption selectivity, and the capacity characteristic of the polymer was investigated by a conventional batch adsorption test and Scatchard plot analysis. The results indicated that coated polymers had specific adsorption to PTL as compared with its co-occurring 5-hydroxymethyl-2-furaldehyde (hydroxymethylfurfural (HMF)), at the same bulk concentration for solution of PTL and HMF, the maximum absorbance in the solid-phase extraction (SPE) method to PTL were 93.97% or 0.654 μg/mg while to HMF they were 76.89% or 0.496 μg/mg. Scatchard analysis revealed that two classes of binding sites were formed in PTL-MIP with dissociation constants of 3.2 × 10−2 and 5.0 × 10−3 mg/mL and the affinity binding sites of 8.029 and 1.364 mg/g, respectively. The recoveries of PTL were more than 90% for the developed MISPE and around 75% for the traditional liquid–liquid extraction in spiked apple juice samples. It was concluded that the method is suitable for the scale-up synthesis of PTL-MIP grafted on silica, and the polymer can be effectively applied as SPE coupled with high-performance liquid chromatography (HPLC) for the determination of PTL in apple juice or other related products.


HPLC chromatograms of loading, washing, and eluting fractions of PTL and HMF from the MIP cartridge. Test samples in each chromatogram from top to bottom: mixed standard of HMF and PTL (10 mg/L), residue solution through the cartridge, first elution and second elution washed with 1 mL of 1% (w/v) HAc aqueous solution.


Molecular imprinted polymers (MIPs) Patulin Mimic Grafting Silica 

Abbreviations used




4,4′-Azobis(4-cyanopentanoic acid)


4,4′-Azobis(4-cyanopentanoic acid) chlorides


Association of Analytical Communities




Coumalic acid




Dimethylamino pyridine


Dimethyl sulfoxide


Elemental analysis


Ethylene glycol dimethacrylate


Fourier transform infrared spectrometry


5-Hydroxymethyl furfural


2-Hydroxynicotinic acid


6-Hydroxynicotinic acid


High-performance liquid chromatography


Isothermal titration calorimetry


Liquid–liquid extraction


Limits of detection


Limits of quantification


Methacrylic acid


Molecularly imprinted polymers


Molecularly imprinted solid-phase extraction


Mass spectrometry


Non-template imprinted polymers


Photodiode array detector




Relative standard deviation


Scanning electron microscope


Solid-phase extraction



We gratefully acknowledge financial support from the Science and Technology Commission of Shanghai Municipality, project no. 06DZ05128.

Supplementary material

216_2011_5282_MOESM1_ESM.pdf (58 kb)
Fig. S1 The Hyperchem-derived energy minimized structure of AM and PTL or analogues. The presence of hydrogen bonds is indicated by dashed lines. Nitrogen = green, carbon = red, oxygen = blue, hydrogen = black. (PDF 57.5 kb)


  1. 1.
    Brian PW, Elson GW, Lowe D (1956) Production of patulin in apple fruits by Penicillium expansum. Nature 178(4527):263–264CrossRefGoogle Scholar
  2. 2.
    Damoglou AP, Campbell DS, Button JE (1985) Some factors governing the production of patulin in apples. Food Microbiology 2(1):3–10CrossRefGoogle Scholar
  3. 3.
    Scott WT, Bullerman LB (1975) Patulin: a mycotoxin of potential concern in foods. J Milk Food Technol 38:695–705Google Scholar
  4. 4.
    WHO (1995) World Health Organization, 44th Report of the Joint FAO/WHO Expert Committee on Food Additives. Technical Report Series 859, p 36Google Scholar
  5. 5.
    FAO (1996) Worldwide regulations for mycotoxins. FAO and Nutrition Paper No. 64.Google Scholar
  6. 6.
    Communities TCotE (2006) No.1881/2006- L364/17. Official Journal European Union:13–14.Google Scholar
  7. 7.
    da Silva SJN, Schuch PZ, Bernard CR, Vainstein MH, Jablonski A, Bender RJ (2007) Patulin in food: state-of-the-art and analytical trends. Revista Brasileira De Fruticultura 29(2):406–413CrossRefGoogle Scholar
  8. 8.
    de Champdore M, Bazzicalupo P, De Napoli L, Montesarchio D, Di Fabio G, Cocozza I, Parracino A, Rossi M, D'Auria S (2007) A new competitive fluorescence assay for the detection of patulin toxin. Anal Chem 79(2):751–757CrossRefGoogle Scholar
  9. 9.
    Brause AR, Trucksess MW, Thomas FS, Page SW (1996) Determination of patulin in apple juice by liquid chromatography: collaborative study. J AOAC Int 79(2):451–455Google Scholar
  10. 10.
    Sewram V, Nair JJ, Nieuwoudt TW, Leggott NL, Shephard GS (2000) Determination of patulin in apple juice by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. J Chromatogr A 897(1–2):365–374CrossRefGoogle Scholar
  11. 11.
    Pichon V, Chapuis-Hugon F (2008) Role of molecularly imprinted polymers for selective determination of environmental pollutants—a review. Anal Chim Acta 622(1–2):48–61CrossRefGoogle Scholar
  12. 12.
    Nováková L, Vlcková H (2009) A review of current trends and advances in modern bio-analytical methods: chromatography and sample preparation. Anal Chim Acta 656(1–2):8–35CrossRefGoogle Scholar
  13. 13.
    Turiel E, Martin-Esteban A (2010) Molecularly imprinted polymers for sample preparation: a review. Anal Chim Acta 668(2):87–99CrossRefGoogle Scholar
  14. 14.
    Baggiani C, Anfossi L, Giovannoli C (2007) Solid phase extraction of food contaminants using molecular imprinted polymers. Anal Chim Acta 591(1):29–39CrossRefGoogle Scholar
  15. 15.
    Weiss R, Freudenschuss M, Krska R, Mizaikoff B (2003) Improving methods of analysis for mycotoxins: molecularly imprinted polymers for deoxynivalenol and zearalenone. Food Add Contam 20(4):386–395CrossRefGoogle Scholar
  16. 16.
    Urraca JL, Marazuela MD, Moreno-Bondi MC (2006) Molecularly imprinted polymers applied to the clean-up of zearalenone and alpha-zearalenol from cereal and swine feed sample extracts. Anal Bioanal Chem 385(7):1155–1161CrossRefGoogle Scholar
  17. 17.
    Urraca JL, Marazuela MD, Merino ER, Orellana G, Moreno-Bondi MC (2006) Molecularly imprinted polymers with a streamlined mimic for zearalenone analysis. J Chromatogr A 1116(1–2):127–134CrossRefGoogle Scholar
  18. 18.
    Baggiani C, Giraudi G, Vanni A (2001) A molecular imprinted polymer with recognition properties towards the carcinogenic mycotoxin ochratoxin A. Bioseparation 10(6):389–394CrossRefGoogle Scholar
  19. 19.
    Jodlbauer J, Maier NM, Lindner W (2002) Towards ochratoxin A selective molecularly imprinted polymers for solid-phase extraction. J Chromatogr A 945(1–2):45–63CrossRefGoogle Scholar
  20. 20.
    Maier NM, Buttinger G, Welhartizki S, Gavioli E, Lindner W (2004) Molecularly imprinted polymer-assisted sample clean-up of ochratoxin A from red wine: merits and limitations. J Chromatogr B 804(1):103–111CrossRefGoogle Scholar
  21. 21.
    Turner NW, Piletska EV, Karim K, Whitcombe M, Malecha M, Magan N, Baggiani C, Piletsky SA (2004) Effect of the solvent on recognition properties of molecularly imprinted polymer specific for ochratoxin A. Biosens Bioelectron 20(6):1060–1067CrossRefGoogle Scholar
  22. 22.
    Appell M, Kendra DF, Kim EK, Maragos CM (2007) Synthesis and evaluation of molecularly imprinted polymers as sorbents of moniliformin. Food Addit Contam 24(1):43–52CrossRefGoogle Scholar
  23. 23.
    Khorrami A, Taherkhani M (2011) Synthesis and evaluation of a molecularly imprinted polymer for pre-concentration of patulin from apple juice. Chromatographia 73:151–156CrossRefGoogle Scholar
  24. 24.
  25. 25.
    Zayats M, Lahav M, Kharitonov AB, Willner I (2002) Imprinting of specific molecular recognition sites in inorganic and organic thin layer membranes associated with ion-sensitive field-effect transistors. Tetrahedron 58(4):815–824CrossRefGoogle Scholar
  26. 26.
    Harrison MA (1988) Presence and stability of patulin in apple products: a review. Journal of Food Safety 9(3):147–153CrossRefGoogle Scholar
  27. 27.
    Farrington K, Magner E, Regan F (2006) Predicting the performance of molecularly imprinted polymers: Selective extraction of caffeine by molecularly imprinted solid phase extraction. Anal Chim Acta 566(1):60–68CrossRefGoogle Scholar
  28. 28.
    Boven G, Oosterling MLCM, Challa G, Jan Schouten A (1990) Grafting kinetics of poly(methyl methacrylate) on microparticulate silica. Polymer 31(12):2377–2383CrossRefGoogle Scholar
  29. 29.
    Sulitzky C, Rückert B, Hall AJ, Lanza F, Unger K, Sellergren B (2001) Grafting of molecularly imprinted polymer films on silica supports containing surface-bound free radical initiators. Macromolecules 35(1):79–91CrossRefGoogle Scholar
  30. 30.
    Sellergren B, Shea KJ (1993) Chiral ion-exchange chromatography. Correlation between solute retention and a theoretical ion-exchange model using imprinted polymers. J Chromatogr A 654(1):17–28CrossRefGoogle Scholar
  31. 31.
    Wu RN, Dang YL, Niu L, Hu H (2008) Application of matrix solid-phase dispersion-HPLC method to determine patulin in apple and apple juice concentrate. Journal of Food Composition and Analysis 21(7):582–586CrossRefGoogle Scholar
  32. 32.
    Yu C, Mosbach K (1997) Molecular imprinting utilizing an amide functional group for hydrogen bonding leading to highly efficient polymers. J Org Chem 62(12):4057–4064CrossRefGoogle Scholar
  33. 33.
    Delaunay-Bertoncini N, Pichon V, Hennion MC (2003) Experimental comparison of three monoclonal antibodies for the class-selective immunoextraction of triazines—correlation with molecular modeling and principal component analysis studies. J Chromatogr A 999(1–2):3–15CrossRefGoogle Scholar
  34. 34.
    Weber A, Dettling M, Brunner H, Tovar GEM (2002) Isothermal titration calorimetry of molecularly imprinted polymer nanospheres. Macromol Rapid Commun 23(14):824–828CrossRefGoogle Scholar
  35. 35.
    Unger KK (1990) Adsorbents in column liquid chromatography, vol 47. Marcel Dekker, New YorkGoogle Scholar
  36. 36.
    Roach JAG, Brause AR, Eisele TA, Rupp HS (2002) HPLC detection of patulin in apple juice with GC/MS confirmation of patulin identity. In: DeVries JW, Trucksess MW, Jackson LS (eds) Advances in experimental medicine and biology: mycotoxins and food safety, vol 504. Kluwer, New York, pp 135–140CrossRefGoogle Scholar
  37. 37.
    Moukas A, Panagiotopoulou V, Markaki P (2008) Determination of patulin in fruit juices using HPLC-DAD and GC-MSD techniques. Food Chem 109(4):860–867CrossRefGoogle Scholar
  38. 38.
    Schebb N, Faber H, Maul R, Heus F, Kool J, Irth H, Karst U (2009) Analysis of glutathione adducts of patulin by means of liquid chromatography (HPLC) with biochemical detection (BCD) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). Anal Bioanal Chem 394(5):1361–1373CrossRefGoogle Scholar
  39. 39.
    Pan M, Wang J, Fang G, Tang W, Wang S (2010) Synthesis and characterization of a molecularly imprinted polymer and its application as SPE enrichment sorbent for determination of trace methimazole in pig samples using HPLC-UV. J Chromatogr B 878(19):1531–1536CrossRefGoogle Scholar
  40. 40.
    Kadakal C, Nas S (2002) Effect of activated charcoal on patulin, fumaric acid and some other properties of apple juice. Nahrung-Food 46(1):31–33CrossRefGoogle Scholar
  41. 41.
    Umpleby RJ, Baxter SC, Rampey AM, Rushton GT, Chen YZ, Shimizu KD (2004) Characterization of the heterogeneous binding site affinity distributions in molecularly imprinted polymers. J Chromatogr B 804(1):141–149CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Dayun Zhao
    • 1
    • 2
    Email author
  • Jingfu Jia
    • 1
  • Xuelei Yu
    • 1
  • Xiangjun Sun
    • 1
    • 2
  1. 1.Department of Food Science and Technology, School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Bor S. Luh Food Safety Research CenterShanghai Jiao Tong UniversityShanghaiChina

Personalised recommendations