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

Research output: Book chapter/Published conference paperConference paper

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Abstract

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
Pages4
Number of pages1
Publication statusUnpublished - 2019
EventAustralian Grasslands Association: Technical Symposium - Australian National University, Canberra, Australia
Duration: 15 May 201316 May 2013
https://www.mla.com.au/research-and-development/search-rd-reports/final-report-details/Productivity-On-Farm/Perennial-Grasses-in-Pasture-Production-Systems-The-2013-Symposium-by-the-Australian-Grasslands-Association/260 (Conference information)

Conference

ConferenceAustralian Grasslands Association
Abbreviated titlePerennial Grasses in Pasture Production Systems
CountryAustralia
CityCanberra
Period15/05/1316/05/13
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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Alkaloids
Methylene Chloride
Electrospray ionization
Liquid chromatography
Solvent extraction
Mass spectrometry
Fluorescence
lolitrem B

Cite this

Weston, P., Quinn, J., & Weston, L. (2019). An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass. Unpublished. In C. Harris (Ed.), Perennial Grasses in Pasture Production Systems (pp. 4). Australia: CSIRO Publishing.
Weston, Paul ; Quinn, Jane ; Weston, Leslie. / An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass. Perennial Grasses in Pasture Production Systems. editor / Carole Harris. Australia : CSIRO Publishing, 2019. pp. 4
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abstract = "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.",
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Weston, P, Quinn, J & Weston, L 2019, An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass. in C Harris (ed.), Perennial Grasses in Pasture Production Systems. CSIRO Publishing, Australia, pp. 4, Australian Grasslands Association, Canberra, Australia, 15/05/13.

An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass. / Weston, Paul; Quinn, Jane; Weston, Leslie.

Perennial Grasses in Pasture Production Systems. ed. / Carole Harris. Australia : CSIRO Publishing, 2019. p. 4.

Research output: Book chapter/Published conference paperConference paper

TY - GEN

T1 - An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass

AU - Weston, Paul

AU - Quinn, Jane

AU - Weston, Leslie

N1 - Imported on 03 May 2017 - DigiTool details were: publisher = Australia: CSIRO, 2013. editor/s (773b) = Carole Harris; Event dates (773o) = May 15-16, 2013; Parent title (773t) = Technical Symposium by Australian Grasslands Association.

PY - 2019

Y1 - 2019

N2 - 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.

AB - 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.

KW - Open access version available

KW - Analytical chemistry

KW - Livestock production

KW - Ryegrass staggers

M3 - Conference paper

SP - 4

BT - Perennial Grasses in Pasture Production Systems

A2 - Harris, Carole

PB - CSIRO Publishing

CY - Australia

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Weston P, Quinn J, Weston L. An improved method for recovering and quantifying neurotoxic alkaloids from endophyte-infected ryegrass. In Harris C, editor, Perennial Grasses in Pasture Production Systems. Australia: CSIRO Publishing. 2019. p. 4