Deakin University
Browse

File(s) under permanent embargo

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

journal contribution
posted on 2017-03-01, 00:00 authored by Esme Candish, Aminreza Khodabandeh, Marianne Gaborieau, Thomas Rodemann, Robert ShellieRobert Shellie, Andrew A Gooley, Emily F Hilder
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.

History

Journal

Analytical and bioanalytical chemistry

Volume

409

Pagination

2189-2199

Location

Berlin, Germany

ISSN

1618-2642

eISSN

1618-2650

Language

eng

Publication classification

C1 Refereed article in a scholarly journal

Copyright notice

2017, Springer-Verlag Berlin Heidelberg

Issue

8

Publisher

Springer Verlag