title: High-resolution fractionation after gas chromatography for effect-directed analysis.
abstract: This research presents an analytical technology for highly efficient, high-resolution, and high-yield fractionation of compounds after gas chromatography (GC) separations. The technology is straightforward, does not require sophisticated cold traps or adsorbent traps, and allows collecting large numbers of fractions during a GC run. The technology is based on direct infusion of a carrier solvent at the end of the GC column, where infusion takes place in the GC oven. Pentane and hexane used as carrier solvent showed good results. Acetonitrile also showed good results as a more polar carrier solvent. Development and optimization of the technology is described, followed by demonstration in a high-throughput effect directed analysis setting toward dioxin receptor bioactivity. The GC fractionation setup was capable of collecting fractions in the second range. As a result, fractionated compounds could be collected into one or two fractions when 6.5 s resolution fractionation was performed. Subsequently, mixtures containing polycyclic aromatic hydrocarbons, of which some are bioactive toward the dioxin receptor, were profiled with a mammalian gene reporter assay. After fractionation into 96-well plates, we used our new approach for direct cell seeding onto the fractions prior to assaying which allowed dioxin receptor bioactivity to be measured directly after fractionation. The current technology represents a great advance in effect directed analysis for environmental screening worldwide as it allows combining the preferred analytical separation technology for often non-polar environmental pollutants with environmentally relevant bioassays, in high resolution.
authors.affiliation: AIMMS Division of BioAnalytical Chemistry, Faculty of Sciences, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Eelco
authors.initials: E
authors.lastname: Pieke
authors.forename: Ferry
authors.initials: F
authors.lastname: Heus
authors.forename: Jorke H
authors.initials: JH
authors.lastname: Kamstra
authors.forename: Marija
authors.initials: M
authors.lastname: Mladic
authors.forename: Martin
authors.initials: M
authors.lastname: van Velzen
authors.forename: Dik
authors.initials: D
authors.lastname: Kamminga
authors.forename: Marja H
authors.initials: MH
authors.lastname: Lamoree
authors.forename: Timo
authors.initials: T
authors.lastname: Hamers
authors.forename: Pim
authors.initials: P
authors.lastname: Leonards
authors.forename: Wilfried M A
authors.initials: WM
authors.lastname: Niessen
authors.forename: Jeroen
authors.initials: J
authors.lastname: Kool
mesh_terms: D000818:Animals; D005591:Chemical Fractionation / Q000379:methods*; D002849:Chromatography, Gas / Q000379:methods*