An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass

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    Most published methods for quantifying neurotoxic alkaloids such as lolitrem B from perennial ryegrass rely on separation using normal phase HPLC followed by fluorescence detection. We report here a method based on reversed-phase HPLC using a C18 column followed by electrospray ionisation liquid chromatography/mass spectrometry (ESI LC-MS). Lolitrem B was easily quantified with this method at very low levels of abundance (limit of detection = 1 pg injected on column). Extraction of dried ryegrass foliage with various solvents showed dichloromethane to be most efficient at recovering lolitrem B, but acetonitrile recovered 75% of the amount extracted by dichloromethane and has numerous advantages over dichlormethane as an extraction solvent. This method should prove to be very useful for quantitating lolitrem B in perennial ryegrass samples because it offers great sensitivity and selectivity and relies on commonly used chromatographic columns and solvents.
    Original languageEnglish
    Title of host publicationPerennial Grasses in Pasture Production Systems
    EditorsCarole Harris
    Place of PublicationAustralia
    PublisherCSIRO Publishing
    Number of pages1
    Publication statusPublished - 2013
    EventAustralian Grasslands Association: Technical Symposium - Australian National University, Canberra, Australia
    Duration: 15 May 201316 May 2013 (Conference information)


    ConferenceAustralian Grasslands Association
    Abbreviated titlePerennial Grasses in Pasture Production Systems
    OtherA symposium held in Canberra during May 2013 on perennial grasses in pasture production systems was the second in a series of regular technical events organised by the Australian Grasslands Association. This technical symposium brought together leading scientists and pasture experts to deliver a structured, broad based review of the main issues affecting perennial grass performance under livestock grazing via a symposium which includes invited review papers on specific topics.
    The project aimed to develop a set of recommendations and priorities for future investment in perennial grass improvement and management for livestock industry investment by the public and private sector, including RDCs. These priorities will be useful for future government and private sector prioritisation of investment into pastures R&D.
    The sentiments established during this symposium are being developed into a paper that will be published along with papers presented in a special edition of CSIRO’s Journal of Crop and Pasture Science, in late 2013. Abstracts of papers presented at the symposium and full poster papers are available in the proceedings of the symposium.
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