An integrated targeted and untargeted approach for the analysis of ergot alkaloids in cereals using UHPLC – hybrid quadrupole time-of-flight mass spectrometry

Arroyo‐Manzanares, Natalia; Mavungu, José Diana Di; Uka, Valdet; Gámiz‐Gracia, Laura; Garcı́a-Campaña, Ana M.; Saeger, Sarah De

doi:10.3920/wmj2015.1900

Cited by 8 publications

(4 citation statements)

References 32 publications

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“…For instance, in our study, speradine A eluted between α-CPA and 2-oxo-CPA, which actually represents the same elution pattern as reported by other researchers [ 33 ]. Similar successful dereplication approaches based on TOF technologies have been reported previously [ 44 , 47 , 48 ].…”

Section: Discussionsupporting

confidence: 80%

Unravelling the Diversity of the Cyclopiazonic Acid Family of Mycotoxins in Aspergillus flavus by UHPLC Triple-TOF HRMS

Uka

Moore

Arroyo‐Manzanares

et al. 2017

Toxins

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Cyclopiazonic acid (α-cyclopiazonic acid, α-CPA) is an indole-hydrindane-tetramic acid neurotoxin produced by various fungal species, including the notorious food and feed contaminant Aspergillus flavus. Despite its discovery in A. flavus cultures approximately 40 years ago, its contribution to the A. flavus mycotoxin burden is consistently minimized by our focus on the more potent carcinogenic aflatoxins also produced by this fungus. Here, we report the screening and identification of several CPA-type alkaloids not previously found in A. flavus cultures. Our identifications of these CPA-type alkaloids are based on a dereplication strategy involving accurate mass high resolution mass spectrometry data and a careful study of the α-CPA fragmentation pattern. In total, 22 CPA-type alkaloids were identified in extracts from the A. flavus strains examined. Of these metabolites, 13 have been previously reported in other fungi, though this is the first report of their existence in A. flavus. Two of our metabolite discoveries, 11,12-dehydro α-CPA and 3-hydroxy-2-oxo CPA, have never been reported for any organism. The conspicuous presence of CPA and its numerous derivatives in A. flavus cultures raises concerns about the long-term and cumulative toxicological effects of these fungal secondary metabolites and their contributions to the entire A. flavus mycotoxin problem.

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Section: Discussionsupporting

confidence: 80%

Unravelling the Diversity of the Cyclopiazonic Acid Family of Mycotoxins in Aspergillus flavus by UHPLC Triple-TOF HRMS

Uka

Moore

Arroyo‐Manzanares

et al. 2017

Toxins

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“… Method 1: Described for the extraction of EAs from rye, based on an extraction with ethyl acetate/MeOH/0.2 mol L ‐1 ammonium bicarbonate pH 8.5 (30 min), phase separation, evaporation, reconstitution in MeOH/MeCN/water, addition of hexane, centrifugation and analysis of the aqueous phase 29 Method 2: Proposed by Guo et al .…”

Section: Resultsmentioning

confidence: 99%

“…Concerning recent occurrence studies about EAs, although the occurrence of main mycotoxins in feed has been extensively studied around the world, 23–26 only a few of these works included EAs 19,26,27 . Moreover, most of the papers including the determination of EAs are focused on cereal grains, 14,28–34 with the inclusion of processed feeds being scarce 19,26,27 . According to the recent available results, the data reported by EFSA in 2017, where a total of 654 feed samples collected in five different countries (Czech Republic, Croatia, Slovenia, the Netherlands and the United Kingdom) between 2011 and 2016, revealed that in more than 50% of these samples, not a single EA was found above the limit of quantification (LOQ).…”

Section: Introductionmentioning

confidence: 99%

Determination of principal ergot alkaloids in swine feeding

et al. 2021

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BACKGROUND Ergot alkaloids are secondary metabolites produced by fungi in the genus Claviceps. They contaminate a large variety of cereals, such as rye, triticale, wheat and barley. The ingestion of contaminated cereals might cause adverse health effects in humans and animals. In fact, pigs, cattle, sheep, and poultry are involved in sporadic outbreaks and, although there are several studies about occurrence of ergot alkaloids in grain cereals, there are scarce studies focused on compound feed. RESULTS Twelve ergot alkaloids have been quantified in 228 feed samples intended for swine. The analytes were extracted using QuEChERS with Z‐Sep+ as sorbent in the clean‐up step, which reduced the matrix effect, allowing limits of quantification between 2.1 and 21.7 μg kg–1. The analytes were subsequently quantified by ultra‐high‐performance liquid chromatography with tandem mass spectrometry (UHPLC–MS/MS). A total of 29 samples (12.7%) revealed contamination by at least one ergot alkaloid, and among contaminated samples, 65% were contaminated by more than one. Only 6 of 12 target ergot alkaloids showed concentrations above the limit of quantification. The concentrations for individual ergot alkaloids ranged between 5.9 μg kg–1 for ergosinine to 145.3 μg kg–1 for ergometrine (the predominant ergot alkaloid), while the total ergot alkaloid content ranged from 5.9 to 158.7 μg kg–1. CONCLUSIONS The occurrence of ergot alkaloids in feed samples in Spain seems to be lower than in other regions of Europe. All the samples fulfilled current recommendations of the feed industry about practical limits for ergot alkaloids in pig feeds. This suggests that the feeds are safe for pig consumption, regarding the presence of ergot alkaloids. © 2021 Society of Chemical Industry.

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“…The methodology applied here can be used in the study of other fungal biosynthetic genes, allowing to identify new compounds that could be of importance in food and feed safety and to assign the genes that are responsible for the production of known fungal secondary metabolites, including mycotoxins. In this regard, the use of molecular networking as a complement de-replication strategy (Yang et al 2013) can be explored, exploiting MS/MS spectra generated in the present study and in previous works Arroyo-Manzanares et al 2015). …”

Section: Discussionmentioning

confidence: 97%

Use of UHPLC high-resolution Orbitrap mass spectrometry to investigate the genes involved in the production of secondary metabolites inAspergillus flavus

Arroyo‐Manzanares

Mavungu

Uka

et al. 2015

Food Additives & Contaminants: Part A

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The fungus Aspergillus flavus is known for its ability to produce the toxic and carcinogenic aflatoxins in food and feed. While aflatoxins are of most concern, A. flavus is predicted to be capable of producing many more metabolites based on a study of its complete genome sequence. Some of these metabolites could be of great importance in food and feed safety. Here we describe an analytical methodology based on Orbitrap HRMS technology that allows the untargeted determination of fungal metabolites, in support of the study of the function of genes involved in secondary metabolism in fungi. The applied strategy implies the detection and identification of differentially expressed metabolites in extracts of wild-type and mutant fungal strains, using Orbitrap high-resolution mass spectrometry (HRMS) accurate mass data. The suitability of this approach was demonstrated by the confirmation of previously characterised genes involved in the aflatoxin biosynthetic pathway, namely a polyketide synthase (pksA), an oxidoreductase (ordA) and a methyltransferase (omtA) gene. Subsequently, the proposed methodology was applied for the detection and identification of metabolites produced by a yet uncharacterised gene cluster in A. favus, cluster 23. Comparative Orbitrap HRMS analysis of extracts of A. flavus wild-type strain and an over-expression mutant for the transcription factor of gene cluster 23 (lepE) demonstrated that this gene cluster is responsible for the production a set of 2-pyridone derivatives, the leporins. Besides the known derivatives leporin B and leporin B precursor that could be identified by automatic de-replication of the accurate mass data, five other compounds belonging to this class of fungal secondary metabolites were detected and identified for the first time, combining MS and multiple-stage MS data.

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An integrated targeted and untargeted approach for the analysis of ergot alkaloids in cereals using UHPLC – hybrid quadrupole time-of-flight mass spectrometry

Abstract: Saeger S. (2015). An integrated targeted and untargeted approach for the analysis of ergot alkaloids in cereals using UHPLC -hybrid quadrupole time-of-flight MS. World Mycotoxin Journal 8 (5)

Cited by 8 publications

References 32 publications

Unravelling the Diversity of the Cyclopiazonic Acid Family of Mycotoxins in Aspergillus flavus by UHPLC Triple-TOF HRMS

Unravelling the Diversity of the Cyclopiazonic Acid Family of Mycotoxins in Aspergillus flavus by UHPLC Triple-TOF HRMS

Determination of principal ergot alkaloids in swine feeding

Use of UHPLC high-resolution Orbitrap mass spectrometry to investigate the genes involved in the production of secondary metabolites inAspergillus flavus

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