Skip to main content
SpringerLink
Log in

Poly(ethylene glycol) functionalization of monolithic poly(divinyl benzene) for improved miniaturized solid phase extraction of protein-rich samples

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Non-specific protein adsorption on hydrophobic solid phase extraction (SPE) adsorbents can reduce the efficacy of purification. To improve sample clean-up, poly(divinyl benzene) (PDVB) monoliths grafted with hydrophilic polyethylene glycol methacrylate (PEGMA) were developed. Residual vinyl groups (RVGs) of the PDVB were employed as anchor points for PEGMA grafting. Two PEGMA monomers, M n 360 and 950, were compared for graft solutions containing 5–20% monomer. Protein binding was qualitatively screened using fluorescently labeled human serum albumin (HSA) to determine optimal PEGMA concentration. The fluorescent signal of PDVB was reduced for PDVB-g-PEGMA360 (10%) and PDVB-g-PEGMA950 (20%). The PEGMA content (w/w%) was quantified by solid state 1H NMR to be 29.9 ± 1.6% for PDVB-g-PEGMA360 and 7.7 ± 1.2% for PDVB-g-PEGMA950. To assess adsorbent performance breakthrough curves for PDVB, PDVB-g-PEGMA360 and PDVB-g-PEGMA950 were compared. The breakthrough volume (V B) and shape of the curve for PDVB-g-PEGMA950 were maintained relative to PDVB (2.3 and 2.8 mL, respectively). A reduced V B of 0.5 mL and shallow breakthrough curve indicated PDVB-g-PEGMA360 was not suitable for SPE. A high ibuprofen recovery of 92 ± 0.30 and 78 ± 0.93% was seen for PDVB and PDVB-g-PEGMA950, respectively. Protein adsorption was reduced from 31 ± 2.41 to 12 ± 0.49% for PDVB and PDVB-g-PEGMA950, respectively. SPE of ibuprofen from plasma was compared for PDVB and PDVB-g-PEGMA950 by at-line electrospray ionization mass spectrometry (ESI-MS). PDVB-g-PEGMA950 demonstrated a threefold increase in assay sensitivity indicating a superior analyte purification.

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. Westerlund D. Direct injection of plasma into column liquid chromatographic systems. Chromatographia. 1987;24:155–64.

    Article  CAS  Google Scholar 

  2. Annesley TM. Ion suppression in mass spectrometry. Clin Chem. 2003;49:1041–4.

    Article  CAS  Google Scholar 

  3. Chen Y, Guo Z, Wang X, Qiu C. Sample preparation. J Chromatogr A. 2008;1184:191–219.

    Article  CAS  Google Scholar 

  4. Li Y, Lee ML. Biocompatible polymeric monoliths for protein and peptide separations. J Sep Sci. 2009;32:3369–78.

    Article  CAS  Google Scholar 

  5. Georgi K, Boos KS. Multidimensional on-line SPE for undisturbed LC-MS-MS analysis of basic drugs in biofluids. LC GC Eur. 2004;17:21–4.

    CAS  Google Scholar 

  6. Cassiano NM, Lima VV, Oliveira RV, de Pietro AC, Cass QB. Development of restricted-access media supports and their application to the direct analysis of biological fluid samples via high-performance liquid chromatography. Anal Bioanal Chem. 2006;384:1462–9.

    Article  CAS  Google Scholar 

  7. Souverain S, Rudaz S, Veuthey J-L. Restricted access materials and large particle supports for on-line sample preparation: an attractive approach for biological fluids analysis. J Chromatogr B. 2004;801:141–56.

    Article  CAS  Google Scholar 

  8. Hagestam IH, Pinkerton TC. Internal surface reversed-phase silica supports for liquid chromatography. Anal Chem. 1985;57:1757–63.

    Article  CAS  Google Scholar 

  9. Cook SE, Pinkerton TC. Characterization of internal surface reversed-phase silica supports for liquid chromatography. J Chromatogr A. 1986;368:233–48.

    Article  CAS  Google Scholar 

  10. Turson M, Zhou M, Jiang P, Dong X. Monolithic poly(ethylhexyl methacrylate-co-ethylene dimethacrylate) column with restricted access layers prepared via reversible addition-fragmentation chain transfer polymerization. J Sep Sci. 2011;34:127–34.

    Article  CAS  Google Scholar 

  11. Candish E, Wirth H-J, Gooley AA, Shellie RA, Hilder EF. Characterization of large surface area polymer monoliths and their utility for rapid, selective solid phase extraction for improved sample clean up. J Chromatogr A. 2015;1410:9–18.

    Article  CAS  Google Scholar 

  12. Yang H, Ulbricht M. Synthesis and performance of PP microfiltration membranes grafted with polymer layers of different structure. Macromol Mater Eng. 2008;293:419–27.

    Article  CAS  Google Scholar 

  13. Gisch DJ, Hunter BT, Feibush B. Shielded hydrophobic phase: a new concept for direct injection analysis of biological fluids by high-performance liquid chromatography. J Chromatogr B. 1988;433:264–8.

    Article  CAS  Google Scholar 

  14. Kanda T, Shirota O, Ohtsu Y, Yamaguchi M. Synthesis and characterization of polymer-coated mixed functional stationary phases with several different hydrophobic groups for direct analysis of biological samples by liquid chromatography. J Chromatogr A. 1996;722:115–21.

    Article  CAS  Google Scholar 

  15. Buchmeiser MR. Polymeric monolithic materials: syntheses, properties, functionalization and applications. Polymer. 2007;48:2187–98.

    Article  CAS  Google Scholar 

  16. Smirnov KN, Dyatchkov IA, Telnov MV, Pirogov AV, Shpigun OA. Effect of monomer mixture composition on structure and chromatographic properties of poly(divinylbenzene-co-ethylvinylbenzene-co-2-hydroxyethyl methacrylate) monolithic rod columns for separation of small molecules. J Chromatogr A. 2011;1218:5010–9.

    Article  CAS  Google Scholar 

  17. Gu B, Armenta JM, Lee ML. Preparation and evaluation of poly (polyethylene glycol methyl ether acrylate-co-polyethylene glycol diacrylate) monolith for protein analysis. J Chromatogr A. 2005;1079:382–91.

    Article  CAS  Google Scholar 

  18. Krenkova J, Lacher NA, Svec F. Highly efficient enzyme reactors containing trypsin and endoproteinase LysC immobilized on porous polymer monolith coupled to MS suitable for analysis of antibodies. Anal Chem. 2009;81:2004–12.

    Article  CAS  Google Scholar 

  19. Barlow KJ, Hao X, Hughes TC, Hutt OE, Polyzos A, Turner KA, et al. Porous, functional, poly (styrene-co-divinylbenzene) monoliths by RAFT polymerization. Polym Chem. 2014;5:722–32.

    Article  CAS  Google Scholar 

  20. Peters EC, Svec F, Fréchet JMJ, Viklund C, Irgum K. Control of porous properties and surface chemistry in “molded” porous polymer monoliths prepared by polymerization in the presence of TEMPO. Macromolecules. 1999;32:6377–9.

    Article  CAS  Google Scholar 

  21. Hubbard KL, Finch JA, Darling GD. Epoxidation of the pendant vinylbenzene groups of commercial poly (divinylbenzene-co-ethylvinylbenzene). React Funct Polym. 1999;42:279–89.

    Article  CAS  Google Scholar 

  22. Tripp JA, Needham TP, Ripp EM, Konzman BG, Homnick PJ. A continuous-flow electrophile scavenger prepared by a simple grafting procedure. React Funct Polym. 2010;70:414–8.

    Article  CAS  Google Scholar 

  23. Gaborieau M, Nebhani L, Graf R, Barner L, Barner-Kowollik C. Accessing quantitative degrees of functionalization on solid substrates via solid-state NMR spectroscopy. Macromolecules. 2010;43:3868–75.

    Article  CAS  Google Scholar 

  24. Claridge TDW (2009) High-resolution NMR techniques in organic chemistry, Second Edition.

  25. Morcombe CR, Zilm KW. Chemical shift referencing in MAS solid state NMR. J Magn Reson. 2003;162:479–86.

    Article  CAS  Google Scholar 

  26. Sýkora D, Peters EC, Svec F. “Molded” porous polymer monoliths: a novel format for capillary gas chromatography stationary phases. Macromol Mater Eng. 2000;275:42–7.

    Article  Google Scholar 

  27. Candish E, Gooley A, Wirth H-J, Dawes PA, Shellie RA, Hilder EF. A simplified approach to direct SPE‐MS. J Sep Sci. 2012;35:2399–406.

    Article  CAS  Google Scholar 

  28. Jarmalavičienė R, Kornyšova O, Westerlund D, Maruška A. Non-particulate (continuous bed or monolithic) restricted-access reversed-phase media for sample clean-up and separation by capillary-format liquid chromatography. Anal Bioanal Chem. 2003;377:902–8.

    Article  Google Scholar 

  29. Jarmalavičienė R, Kornyšova O, Bendokas V, Westerlund D, Buszewski B, Maruška A. Restricted-access media development for direct analysis of drugs in biofluids using capillary liquid chromatography. Anal Bioanal Chem. 2008;391:2323–8.

    Article  Google Scholar 

  30. D’Sa RA, Meenan BJ. Chemical grafting of poly (ethylene glycol) methyl ether methacrylate onto polymer surfaces by atmospheric pressure plasma processing. Langmuir. 2010;26:1894–903.

    Article  Google Scholar 

  31. Mangiante G, Alcouffe P, Burdin B, Gaborieau M, Zeno E, Petit-Conil M, et al. Green nondegrading approach to alkyne-functionalized cellulose fibers and biohybrids thereof: synthesis and mapping of the derivatization. Biomacromolecules. 2013;14:254–63.

    Article  CAS  Google Scholar 

  32. Bielicka-Daszkiewicz K, Voelkel A. Theoretical and experimental methods of determination of the breakthrough volume of SPE sorbents. Talanta. 2009;80:614–21.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

RAS is the recipient of an Australian Research Council Australian Research Fellowship (DP110104923). EC is the recipient of an Australian Postgraduate Award. Support from the University of Tasmania Central Science Laboratory is gratefully acknowledged. We gratefully acknowledge Dr. Karsten Gömann and Dr. Sandrin Feig (University of Tasmania) for assistance with scanning electron microscopy and Patrice Castignolles (University of Western Sydney) for helpful discussions.

Author information

Author notes

  1. Emily F. Hilder

    Present address: Future Industries Institute, University of South Australia, GPO Box 2471, Adelaide, South Australia, 5001, Australia

Authors and Affiliations

  1. Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart, Tasmania, 7001, Australia

    Esme Candish, Aminreza Khodabandeh, Robert A. Shellie, Andrew A. Gooley & Emily F. Hilder

  2. Trajan Scientific & Medical, 7 Argent Place, Ringwood, Victoria, 3134, Australia

    Esme Candish & Andrew A. Gooley

  3. Molecular Medicine Research Group, Australian Centre for Research on Separation Science, School of Science and Health, Western Sydney University, Locked Bag 1797, Penrith, New South Wales, 2751, Australia

    Marianne Gaborieau

  4. Central Science Laboratory, University of Tasmania, Private Bag 74, Hobart, Tasmania, 7001, Australia

    Thomas Rodemann

Authors
  1. Esme Candish
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aminreza Khodabandeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marianne Gaborieau
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thomas Rodemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert A. Shellie
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Andrew A. Gooley
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Emily F. Hilder
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Emily F. Hilder.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest

Ethics approval

This study was approved by the Tasmanian Health and Medical Human Research Ethics Committee (protocol H0010488), Australia. Following this protocol, blood samples were obtained from participants with informed consent.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 5304 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Candish, E., Khodabandeh, A., Gaborieau, M. et al. Poly(ethylene glycol) functionalization of monolithic poly(divinyl benzene) for improved miniaturized solid phase extraction of protein-rich samples. Anal Bioanal Chem 409, 2189–2199 (2017). https://doi.org/10.1007/s00216-016-0164-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-016-0164-y

Keywords

Search

Navigation