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

Toxins

Simone Vassiliadis¹  Kathryn M. Guthridge¹  Simone J. Rochfort¹  Aaron C. Elkins¹  German C. Spangenberg¹  Priyanka Reddy¹ 

[1] Agriculture Victoria, AgriBio Centre for AgriBioscience, Bundoora, VIC 3083, Australia;

关键词: epichloë endophyte;    lolium perenne l.;    ryegrass;    alkaloid;    mycotoxins;    quantitation;    liquid chromatography–mass spectrometry;   

DOI  :  10.3390/toxins11110649

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown