Construction of polymer monolithic columns in polypropylene ink-pen tubes for separation of proteins by cation-exchange chromatography

Author(s)

Henrique do Nascimento, Fernando ^[1] ; Trazzi, Catharina Raquel Lemos ^[1] ; Moraes, Amanda Hanashiro ^[1] ; Velasques, Caryna Moraes ^[1] ; Costa, Diego Miranda de Souza ^[1] ; Masini, Jorge Cesar ^[1]

Affiliation

^[1] Univ Sao Paulo, Inst Quim, Dept Quim Fundamental, Ave Prof Lineu Prestes 748, BR-05508000 Sao Paulo - Brazil

• Document type: Journal article
• Source: JOURNAL OF SEPARATION SCIENCE; v. 43, n. 22 OCT 2020.
• Web of Science Citations: 1

Abstract

We describe the synthesis of polymer monoliths inside polypropylene tubes from ink pens. These tubes are cheap, chemically stable, and resistant to pressure. UV-initiated grafting with 5 wt% benzophenone in methanol for 20 min activated the internal surface, thus enabling the covalent binding of ethylene glycol dimethacrylate, also via photografting. The pendant vinyl groups attached a poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) monolith prepared via photopolymerization. These tubes measured 100-110 mm long, with 2 mm of internal diameter. The parent monoliths were functionalized with Na2SO3 or iminodiacetate to produce strong and weak cation exchangers, respectively. The columns exhibited permeabilities varying from 2.7 to 3.3 x 10(-13) m(2), which enabled the separation of proteins at 500 mu L/min and back pressures <2.8 MPa. Neither structure collapse nor monolith detachment occurred at flow rates as high as 2.0 mL/min, which produced back pressures between 6.9 and 9.0 MPa. The retention times of ovalbumin, ribonuclease A, cytochrome C, and lysozyme in salt gradient at pH 7.0 followed the order of increasing isoelectric points, confirming the cation exchange mechanism. Separation and determination of lysozyme in egg white proved the applicability of the columns to the analysis of complex samples. (AU)

• FAPESP's process: 13/18507-4 - Porous monolithic materials for chromatographic separations and sample preparation
• Grantee: Jorge Cesar Masini
• Support Opportunities: Regular Research Grants