title: Phosphatidylcholine covalently linked to a methacrylate-based monolith as a biomimetic stationary phase for capillary liquid chromatography.
abstract: In this study a strategy to immobilize phospholipids onto a polymer-based stationary phase is described. Methacrylate-based monoliths in capillary format (150×0.1mm) were modified by soybean phosphatidylcholine through 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide coupling to obtain stationary phases suitable to mimic cell surface membranes. The covalent coupling reaction involves the phosphate group in phospholipids; therefore, the described methodology is suitable for all types of phospholipids. Immobilization of soy bean phosphatidylcholine on the monolith was confirmed by attenuated total reflectance Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry of the fatty alcohol profile, generated upon reductive cleavage of the fatty acyl side chains of the phospholipid on the monolith surface with lithium aluminium hydride. The prepared stationary phases were evaluated through studies on the retention of low-molar mass model analytes including neutral, acidic, and basic compounds. Liquid chromatographic studies confirmed predominant hydrophobic interactions between the analytes and the synthesized stationary phase; however, electrostatic interactions contributed to the retention as well. The synthesized columns showed high stability even with fully aqueous mobile phases such as Dulbecco's phosphate-buffered saline solution.
authors.affiliation: Institute of Analytical Chemistry of the ASCR, v. v. i., Veveří 97, 60200 Brno, Czech Republic. Electronic address: moravcova@iach.cz.
authors.forename: Dana
authors.initials: D
authors.lastname: Moravcová
authors.affiliation: Department of Analytical Chemistry, University of Valencia, Dr. Moliner 50, 46100 Burjassot, Valencia, Spain.
authors.forename: Enrique Javier
authors.initials: EJ
authors.lastname: Carrasco-Correa
authors.affiliation: Institute of Analytical Chemistry of the ASCR, v. v. i., Veveří 97, 60200 Brno, Czech Republic.
authors.forename: Josef
authors.initials: J
authors.lastname: Planeta
authors.affiliation: Institute of Pharmaceutical Sciences, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lämmerhofer
authors.affiliation: Department of Chemistry, P.O. Box 55, 00014 University of Helsinki, Finland. Electronic address: susanne.wiedmer@helsinki.fi.
authors.forename: Susanne K
authors.initials: SK
authors.lastname: Wiedmer
mesh_terms: D032701:Biomimetics; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation* / Q000379:methods*; D002853:Chromatography, Liquid / Q000295:instrumentation*; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D008689:Methacrylates / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D008689:Methacrylates; D010713:Phosphatidylcholines; D010743:Phospholipids; D011108:Polymers; D014867:Water; D005022:Ethyldimethylaminopropyl Carbodiimide
keywords: ATR-FTIR; Analyte–membrane interaction; Capillary LC; Immobilized artificial membrane; Monolithic column; Phospholipids