title: LC-method development for the quantification of neuromedin-like peptides. Emphasis on column choice and mobile phase composition.
abstract: In this study, the separation of four neuromedin-like peptides is investigated on four different core-shell stationary phases. Moreover, the effect of the mobile phase composition, i.e. organic modifier (acetonitrile and methanol) and additive (trifluoroacetic acid, formic acid, acetic acid, ammonium formate and ammonium acetate) on the chromatographic performance is studied. An improvement in chromatographic performance is observed when using the ammonium salt instead of its corresponding acid as additive, except for the column containing a positively charged surface (C18+). In general, the RP-Amide column provided the highest separation power with different mobile phases. However, for the neuromedin-like peptides of interest, the C18+ column in combination with a mobile phase containing methanol as organic modifier and acetic acid as additive provided narrower and higher peaks. A three-factor, three-level design is applied to further optimize the method in terms of increased peak height and reduced solvent consumption, without loss in resolution. The optimized method was subsequently used to assess the in vitro microdialysis recovery of the peptides of interest. Recovery values between 4 and 8% were obtained using a perfusion flow rate of 2μL/min.
authors.affiliation: Department of Pharmaceutical Chemistry, Drug Analysis and Drug Information (FASC), Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: yannick.van.wanseele@vub.ac.be.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Van Wanseele
authors.affiliation: Department of Analytical Chemistry and Pharmaceutical Technology (FABI), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: johan.viaene@vub.ac.be.
authors.forename: Johan
authors.initials: J
authors.lastname: Viaene
authors.affiliation: Department of Pharmaceutical Chemistry, Drug Analysis and Drug Information (FASC), Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: leslievdborre@gmail.com.
authors.forename: Leslie
authors.initials: L
authors.lastname: Van den Borre
authors.affiliation: Department of Pharmaceutical Chemistry, Drug Analysis and Drug Information (FASC), Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: kathleen_dewachter@hotmail.com.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Dewachter
authors.affiliation: Department of Analytical Chemistry and Pharmaceutical Technology (FABI), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: yvanvdh@vub.ac.be.
authors.forename: Yvan
authors.initials: Y
authors.lastname: Vander Heyden
authors.affiliation: Department of Pharmaceutical Chemistry, Drug Analysis and Drug Information (FASC), Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: ilse.smolders@vub.ac.be.
authors.forename: Ilse
authors.initials: I
authors.lastname: Smolders
authors.affiliation: Department of Pharmaceutical Chemistry, Drug Analysis and Drug Information (FASC), Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: aveeckha@vub.ac.be.
authors.forename: Ann
authors.initials: A
authors.lastname: Van Eeckhaut
mesh_terms: D000085:Acetates / Q000737:chemistry; D019342:Acetic Acid / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D005561:Formates / Q000737:chemistry; D000432:Methanol / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D014269:Trifluoroacetic Acid / Q000737:chemistry
chemical_list: D000085:Acetates; D000097:Acetonitriles; D005561:Formates; D010455:Peptides; D012997:Solvents; C030544:formic acid; D014269:Trifluoroacetic Acid; D019342:Acetic Acid; C018824:ammonium acetate; D000432:Methanol; C032159:acetonitrile
keywords: Column screening; Design of experiments; In vitro microdialysis recovery; Mobile phase screening; Neuropeptides