title: Identification and Quantification of Loline-Type Alkaloids in Endophyte-Infected Grasses by LC-MS/MS.
abstract: Lolines, fungal metabolites of the grass-endophyte association, were identified and quantified using newly developed LC-MS/MS methods in endophyte-infected grasses belonging to the Lolium and Festuca genera after extraction with three different solvents using two extraction methods. The shaking extraction method with isopropanol/water was superior to the other methods due to its high sensitivity, high accuracy (recovery within or close to the range of 80-120%), and high precision (coefficient of variation of <10%). Seven loline alkaloids were identified and quantified using our newly established LC-MS/MS methods, and N-formylloline was the most abundant (5 mg/g dry matter), followed by N-acetylloline. These LC-MS/MS methods used the shortest sample handling time and the fewest sample preparation steps and proved to be good alternatives to existing GC and GC-MS analytical methods without compromising analytical efficiency. In conclusion, we developed for the first time a highly sensitive quantitative LC-MS/MS analytical method for the accurate and reproducible quantification and a LightSight-assisted LC-QTRAP/MS qualitative method for the tentative identification of loline-type alkaloids in endophyte-infected grasses.
authors.affiliation: Department of Agroecology, Aarhus University , DK-4200 Slagelse, Denmark.
authors.forename: Khem B
authors.initials: KB
authors.lastname: Adhikari
authors.affiliation: Department of Agroecology, Aarhus University , DK-4200 Slagelse, Denmark.
authors.forename: Birte
authors.initials: B
authors.lastname: Boelt
authors.affiliation: Department of Agroecology, Aarhus University , DK-4200 Slagelse, Denmark.
authors.forename: Inge S
authors.initials: IS
authors.lastname: Fomsgaard
mesh_terms: D000470:Alkaloids / Q000737:chemistry* / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D060026:Endophytes / Q000378:metabolism*; D031751:Festuca / Q000737:chemistry / Q000382:microbiology*; D005658:Fungi / Q000378:metabolism*; D008129:Lolium / Q000737:chemistry / Q000382:microbiology*; D006109:Poaceae / Q000737:chemistry / Q000382:microbiology*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000470:Alkaloids; C059478:loline
keywords: LC-MS; LightSight software; endophytes; lolines