title: Poly(ethylene glycol) functionalization of monolithic poly(divinyl benzene) for improved miniaturized solid phase extraction of protein-rich samples.
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, Mn360 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 state1H NMR to be 29.9 ± 1.6% for PDVB-g-PEGMA360and 7.7 ± 1.2% for PDVB-g-PEGMA950. To assess adsorbent performance breakthrough curves for PDVB, PDVB-g-PEGMA360and PDVB-g-PEGMA950were compared. The breakthrough volume (VB) and shape of the curve for PDVB-g-PEGMA950were maintained relative to PDVB (2.3 and 2.8 mL, respectively). A reduced VBof 0.5 mL and shallow breakthrough curve indicated PDVB-g-PEGMA360was 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-PEGMA950by at-line electrospray ionization mass spectrometry (ESI-MS). PDVB-g-PEGMA950demonstrated a threefold increase in assay sensitivity indicating a superior analyte purification.
authors.affiliation: Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart, Tasmania, 7001, Australia.|Trajan Scientific & Medical, 7 Argent Place, Ringwood, Victoria, 3134, Australia.
authors.forename: Esme
authors.initials: E
authors.lastname: Candish
authors.affiliation: Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart, Tasmania, 7001, Australia.
authors.forename: Aminreza
authors.initials: A
authors.lastname: Khodabandeh
authors.affiliation: 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.
authors.forename: Marianne
authors.initials: M
authors.lastname: Gaborieau
authors.affiliation: Central Science Laboratory, University of Tasmania, Private Bag 74, Hobart, Tasmania, 7001, Australia.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rodemann
authors.affiliation: Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart, Tasmania, 7001, Australia.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Shellie
authors.affiliation: Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart, Tasmania, 7001, Australia.|Trajan Scientific & Medical, 7 Argent Place, Ringwood, Victoria, 3134, Australia.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Gooley
authors.affiliation: Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart, Tasmania, 7001, Australia. emily.hilder@unisa.edu.au.|Future Industries Institute, University of South Australia, GPO Box 2471, Adelaide, South Australia, 5001, Australia. emily.hilder@unisa.edu.au.
authors.forename: Emily F
authors.initials: EF
authors.lastname: Hilder
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008904:Miniaturization; D019906:Nuclear Magnetic Resonance, Biomolecular; D011092:Polyethylene Glycols / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D052616:Solid Phase Extraction / Q000379:methods*; D014753:Vinyl Compounds / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D011108:Polymers; D011506:Proteins; D014753:Vinyl Compounds; D011092:Polyethylene Glycols; C004985:divinyl benzene
keywords: Biocompatible; Grafting; Porous polymer monolith; Sample preparation; Solid phase extraction