Fast construction of polymer monolithic columns inside fluorinated ethylene propylene (FEP) tubes for separation of proteins by reversed-phase liquid chromatography.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: Netherlands NLM ID: 2984816R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-3573 (Electronic) Linking ISSN: 00399140 NLM ISO Abbreviation: Talanta Subsets: MEDLINE
    • Publication Information:
      Publication: Amsterdam : Elsevier
      Original Publication: Oxford : Pergamon Press
    • Subject Terms:
      Muramidase/*isolation & purification ; Myoglobin/*isolation & purification ; Ovalbumin/*isolation & purification ; Polymers/*chemistry ; Polytetrafluoroethylene/*analogs & derivatives ; Ribonuclease, Pancreatic/*isolation & purification; Animals ; Cattle ; Chickens ; Chromatography, Reverse-Phase ; Horses ; Muramidase/chemistry ; Muramidase/metabolism ; Myoglobin/chemistry ; Ovalbumin/chemistry ; Polytetrafluoroethylene/chemistry ; Ribonuclease, Pancreatic/chemistry
    • Abstract:
      This paper describes the preparation of polymer monolithic columns in the confines of fluorinated ethylene propylene (FEP) tubes. These tubes are cheap, chemically stable, and widely used in flow analysis laboratories. UV-initiated grafting with 5 wt% benzophenone in methanol for 1 h activated the internal surface walls, thus enabling the further covalent binding of ethylene glycol dimethacrylate (EDMA) from a 15 wt% solution in methanol, also via photografting. Both steps used 254 nm radiation under a potency of 120 mJ cm 2 . ATR-FTIR measurements revealed the presence of carbonyl, alkyl and vinyl groups in the functionalized FEP. The density of vinyl groups was high enough to firmly attach a poly(lauryl methacrylate-co-ethylene glycol dimethacrylate) monolith in 120 × 1.57 mm i.d. tubes, prepared via photopolymerization. The total preparation lasts less than 2-h. The columns were permeable, (1.58 ± 0.06) × 10 -13  m 2 , providing reproducible chromatographic parameters of retention times, retention factor, selectivity, and resolution. The monoliths were stable at flow rates of 500 μL min -1 , collapsing only at flow rates >700 μL min -1 , a condition that increased the backpressure over 1000 psi (experiments at the room temperature). The separation of proteins by reversed-phase liquid chromatography demonstrated the efficiency of the columns. Determination of egg white proteins (ovalbumin and lysozyme) and myoglobin in spiked urine proved the applicability to the analysis of real samples.
      Competing Interests: Declaration of competing interest The authors declare that there are no competing interests.
      (Copyright © 2020 Elsevier B.V. All rights reserved.)
    • Contributed Indexing:
      Keywords: Egg white; Fluoropolymer tubes; Liquid chromatography; Monolithic columns; Photografting; Urine
    • Accession Number:
      0 (Myoglobin)
      0 (Polymers)
      0 (fluorinated ethylene propylene)
      9002-84-0 (Polytetrafluoroethylene)
      9006-59-1 (Ovalbumin)
      EC 3.1.27.5 (Ribonuclease, Pancreatic)
      EC 3.2.1.17 (Muramidase)
    • Publication Date:
      Date Created: 20200606 Date Completed: 20210126 Latest Revision: 20210126
    • Publication Date:
      20240829
    • Accession Number:
      10.1016/j.talanta.2020.121063
    • Accession Number:
      32498847