Next Article in Journal
Uptake, Growth, and Pigment Changes in Lemna minor L. Exposed to Environmental Concentrations of Cylindrospermopsin
Previous Article in Journal
Crosstalk between Human Microvascular Endothelial Cells and Tubular Epithelial Cells Modulates Pro-Inflammatory Responses Induced by Shiga Toxin Type 2 and Subtilase Cytotoxin
Open AccessArticle

A Simple LC–MS Method for the Quantitation of Alkaloids in Endophyte-Infected Perennial Ryegrass

1
Agriculture Victoria, AgriBio Centre for AgriBioscience, Bundoora, VIC 3083, Australia
2
School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia
*
Author to whom correspondence should be addressed.
Toxins 2019, 11(11), 649; https://doi.org/10.3390/toxins11110649
Received: 3 October 2019 / Revised: 25 October 2019 / Accepted: 1 November 2019 / Published: 7 November 2019
(This article belongs to the Section Mycotoxins)
The rapid identification and quantitation of alkaloids produced by Epichloë endophyte-infected pasture grass is important for the agricultural industry. Beneficial alkaloids, such as peramine, provide the grass with enhanced insect protection. Conversely, ergovaline and lolitrem B can negatively impact livestock. Currently, a single validated method to measure these combined alkaloids in planta does not exist. Here, a simple two-step extraction method was developed for Epichloë-infected perennial ryegrass (Lolium perenne L.). Peramine, ergovaline and lolitrem B were quantified using liquid chromatography–mass spectrometry (LC–MS). Alkaloid linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, selectivity, recovery, matrix effect and robustness were all established. The validated method was applied to eight different ryegrass-endophyte symbiota. Robustness was established by comparing quantitation results across two additional instruments; a triple quadruple mass spectrometer (QQQ MS) and by fluorescence detection (FLD). Quantitation results were similar across all three instruments, indicating good reproducibility. LOQ values ranged from 0.8 ng/mL to 6 ng/mL, approximately one hundred times lower than those established by previous work using FLD (for ergovaline and lolitrem B), and LC–MS (for peramine). This work provides the first highly sensitive quantitative LC–MS method for the accurate and reproducible quantitation of important endophyte-derived alkaloids.
Keywords: Epichloë endophyte; Lolium perenne L.; ryegrass; alkaloid; mycotoxins; quantitation; liquid chromatography–mass spectrometry Epichloë endophyte; Lolium perenne L.; ryegrass; alkaloid; mycotoxins; quantitation; liquid chromatography–mass spectrometry
Show Figures

Graphical abstract

MDPI and ACS Style

Vassiliadis, S.; Elkins, A.C.; Reddy, P.; Guthridge, K.M.; Spangenberg, G.C.; Rochfort, S.J. A Simple LC–MS Method for the Quantitation of Alkaloids in Endophyte-Infected Perennial Ryegrass. Toxins 2019, 11, 649.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop