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Analytical and Bioanalytical Chemistry

, Volume 405, Issue 7, pp 2175–2183 | Cite as

Nanoparticle-modified monolithic pipette tips for phosphopeptide enrichment

  • Jana Krenkova
  • Frantisek Foret
Original Paper

Abstract

We have developed nanoparticle-modified monoliths in pipette tips for selective and efficient enrichment of phosphopeptides. The 5 μL monolithic beds were prepared by UV-initiated polymerization in 200 μL polypropylene pipette tips and either iron oxide or hydroxyapatite nanoparticles were used for monolith modification. Iron oxide nanoparticles were prepared by a co-precipitation method and stabilized by citrate ions. A stable coating of iron oxide nanoparticles on the pore surface of the monolith was obtained via multivalent electrostatic interactions of citrate ions on the surface of nanoparticles with a quaternary amine functionalized poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) monolith. Hydroxyapatite nanoparticles were incorporated into the poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) monolith by simply admixing them in the polymerization mixture followed by in situ polymerization. The nanoparticle-modified monoliths were compared with commercially available titanium dioxide pipette tips. Performance of the developed and commercially available sorbents was demonstrated with the efficient and selective enrichment of phosphopeptides from peptide mixtures of α-casein and β-casein digests followed by off-line MALDI/MS analysis.

Keywords

Nanoparticles Monolith Pipette tip Enrichment Phosphopeptides 

Abbreviations

DHB

2,5-Dihydroxybenzoic acid

DMPAP

2,2-Dimethoxy-2-phenylacetophenone

EDAX

Energy dispersive X-ray analysis

EDMA

Ethylene dimethacrylate

HEMA

2-Hydroxyethyl methacrylate

IMAC

Immobilized metal ion affinity chromatography

META

[3-(Methacryloylamino)propyl]trimethylammonium chloride

MMA

Methyl methacrylate

MOAC

Metal oxide affinity chromatography

SEM

Scanning electron microscopy

Notes

Acknowledgements

The Project is funded from the SoMoPro programme. Research leading to these results has received a financial contribution from the European Community within the Seventh Framework Programme (FP/2007-2013) under Grant Agreement No. 229603. The research is also co-financed by the South Moravian Region. Additional financial support from the Grant Agency of the Czech Republic (P301/11/2055 and P206/12/G014) and the institutional research plan (RVO: 68081715) is also acknowledged.

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Institute of Analytical Chemistry of the Academy of Sciences of the Czech Republic, v. v. iBrnoCzech Republic

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