title: A simple method for simultaneous RP-HPLC determination of indolic compounds related to bacterial biosynthesis of indole-3-acetic acid.
abstract: In this short technical report, we present a fast and simple procedure for sample preparation and a single-run Reversed Phase High Performance Liquid Chromatography (RP-HPLC) determination of seven indoles (indole-3-acetic acid, indole-3-acetamide, indole-3-acetonitrile, indole-3-ethanol, indole-3-lactic acid, tryptamine and tryptophan) in bacterial culture supernatants. The separation of the analytes, after a single centrifugal filtration clean-up step, was performed using a gradient elution on a symmetry C8 column followed by fluorimetric detection (λ(ex) = 280/λ(em) = 350 nm). The calibration curves were linear for all of the studied compounds over the concentration range of 0.0625-125 μg mL(-1) (r ( 2 ) ≥ 0.998) and the limits of detection were below 0.015 μg mL(-1). The applicability of the method was confirmed by analysis of Pseudomonas putida culture supernatants.
authors.affiliation: Department of Biochemistry, Faculty of Agriculture and Biology, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159, 02-776, Warsaw, Poland. michal_szkop@sggw.pl
authors.forename: Michał
authors.initials: M
authors.lastname: Szkop
authors.forename: Wiesław
authors.initials: W
authors.lastname: Bielawski
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007210:Indoleacetic Acids / Q000032:analysis*; D007211:Indoles / Q000032:analysis*; D016958:Pseudomonas putida / Q000378:metabolism*
chemical_list: D007210:Indoleacetic Acids; D007211:Indoles; C030737:indoleacetic acid