title: Liquid-liquid extraction and liquid chromatography-mass spectrometry detection of curcuminoids from bacterial culture medium.
abstract: Liquid chromatography-mass spectrometry (LC-MS) has been used to detect polyphenolic curcuminoids found in turmeric but studies of metabolism by bacterial and mammalian cells in vitro are compromised by poor recovery from the culture medium. We report a liquid-liquid extraction procedure with ethyl acetate and use LC-MS to quantify extracted curcuminoids. Ethyl acetate allows recoveries of ∼ 80-86% of curcuminoids from the bacterial growth medium, bacterial cell lysate and combined bacterial cell and growth medium matrices; a clear improvement over acetonitrile where recoveries were ∼ 25-66%. This optimised method will enable studies of curcuminoid metabolism and may be applicable to other hydrophobic polyphenolic compounds.
authors.affiliation: CSIRO Food, Nutrition and Bioproducts Flagship, 671 Sneydes Road, Werribee, Vic 3030, Australia; Department of Chemical and Biomolecular Engineering, The University of Melbourne, Parkville, Vic 3010, Australia; Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Vic 3010, Australia.
authors.forename: Suryani
authors.initials: S
authors.lastname: Tan
authors.affiliation: Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Vic 3010, Australia; Metabolomics Australia, The University of Melbourne, Parkville, Vic 3010, Australia.
authors.forename: Thusitha W T
authors.initials: TW
authors.lastname: Rupasinghe
authors.affiliation: Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Vic 3010, Australia; Metabolomics Australia, The University of Melbourne, Parkville, Vic 3010, Australia.
authors.forename: Dedreia L
authors.initials: DL
authors.lastname: Tull
authors.affiliation: CSIRO Food, Nutrition and Bioproducts Flagship, 671 Sneydes Road, Werribee, Vic 3030, Australia.
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Augustin
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Melbourne, Parkville, Vic 3010, Australia; Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Vic 3010, Australia; ARC Dairy Innovation Hub, The University of Melbourne, Parkville, Vic 3010, Australia. Electronic address: sgras@unimelb.edu.au.
authors.forename: Sally L
authors.initials: SL
authors.lastname: Gras
mesh_terms: D019149:Bioreactors / Q000382:microbiology; D002853:Chromatography, Liquid / Q000379:methods*; D003474:Curcumin / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D004925:Escherichia / Q000378:metabolism*; D016014:Linear Models; D059625:Liquid-Liquid Extraction / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D059808:Polyphenols / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D059808:Polyphenols; D003474:Curcumin
keywords: Cell medium; Curcumin; Curcuminoids; Ethyl acetate; LC–MS; Recovery