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. 2016 Nov 11;8(11):328.
doi: 10.3390/toxins8110328.

Development and Validation of a LC-ESI-MS/MS Method for the Determination of Alternaria Toxins Alternariol, Alternariol Methyl-Ether and Tentoxin in Tomato and Tomato-Based Products

Yelko Rodríguez-Carrasco  1 , Jordi Mañes  2 , Houda Berrada  3 , Cristina Juan  4
Affiliations

Development and Validation of a LC-ESI-MS/MS Method for the Determination of Alternaria Toxins Alternariol, Alternariol Methyl-Ether and Tentoxin in Tomato and Tomato-Based Products

Yelko Rodríguez-Carrasco et al. Toxins (Basel). .

Abstract

Alternaria species are capable of producing several secondary toxic metabolites in infected plants and in agricultural commodities, which play important roles in food safety. Alternaria alternata turn out to be the most frequent fungal species invading tomatoes. Alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN) are some of the main Alternaria mycotoxins that can be found as contaminants in food. In this work, an analytical method based on liquid chromatography (LC) tandem mass spectrometry (MS/MS) detection for the simultaneous quantification of AOH, AME, and TEN in tomato and tomato-based products was developed. Mycotoxin analysis was performed by dispersive liquid-liquid microextraction (DLLME) combined with LC-ESI-MS/MS. Careful optimization of the MS/MS parameters was performed with an LC/MS system with the ESI interface in the positive ion mode. Mycotoxins were efficiently extracted from sample extract into a droplet of chloroform (100 µL) by DLLME technique using acetonitrile as a disperser solvent. Method validation following the Commission Decision No. 2002/657/EC was carried out by using tomato juice as a blank matrix. Limits of detection and quantitation were, respectively, in the range 0.7 and 3.5 ng/g. Recovery rates were above 80%. Relative standard deviations of repeatability (RSDr) and intermediate reproducibility (RSDR) were ≤ 9% and ≤ 15%, respectively, at levels of 25 and 50 ng/g. Five out of 30 analyzed samples resulted positive to at least one Alternaria toxin investigated. AOH was the most common Alternaria toxin found, but at levels close to LOQ (average content: 3.75 ng/g).

Keywords: Alternaria; LC-MS/MS; dispersive liquid-liquid microextraction; tomato.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
MATLAB-based surface response design showing the influence of AcN and CHCl3 ratio in the extraction efficiency of AOH. Recovery experiments were conducted by spiking tomato extract blank samples with 25 ng/g of each targeted mycotoxin.
Figure 2
Figure 2
Influence of the volume of sample extract in the extraction efficiency of the Alternaria toxins studied. Recovery experiments were conducted by spiking tomato extract blank samples with 25 ng/g of each target mycotoxin.
Figure 3
Figure 3
MRM chromatograms of tomato juice spiked with 3.5 µg/L of AOH, AME, and TEN (corresponding to 1 ng of each injected toxin).
Figure 4
Figure 4
MRM chromatogram of a naturally contaminated tomato sample with AOH at 5.8 ng/g.
See this image and copyright information in PMC

References

    1. Logrieco A., Moretti A., Solfrizzo M. Alternaria Toxins and Plant Diseases: An Overview of Origin, Occurrence and Risks. World Mycotoxin J. 2009;2:129–140. doi: 10.3920/WMJ2009.1145. - DOI
    1. Siciliano C.I., Ortu G., Gilardi G., Gullino M.L., Garibaldi A. Mycotoxin Production in Liquid Culture and on Plants Infected with Alternaria spp. Isolated from Rocket and Cabbage. Toxins. 2015;7:743–754. doi: 10.3390/toxins7030743. - DOI - PMC - PubMed
    1. EFSA Panel on Contaminants in the Food Chain (CONTAM) Scientific Opinion on the Risks for Animal and Public Health Related to the Presence of Alternaria toxins in Feed and Food. EFSA J. 2011 doi: 10.2903/j.efsa.2011.2407. - DOI
    1. Bensassi F., Gallerne C., Dein O.S.E., Hajlaoui M.R., Bacha H., Lemaire C. Mechanism of Alternariol Monomethyl Ether-Induced Mitochondrial Apoptosis in Human Colon Carcinoma Cells. Toxicology. 2011;290:230–240. doi: 10.1016/j.tox.2011.09.087. - DOI - PubMed
    1. Bensassi F., Gallerne C., Sharaf El Dein O., Hajlaoui M.R., Bacha H., Lemaire C. Cell Death Induced by the Alternaria Mycotoxin Alternariol. Toxicol. In Vitro. 2012;26:915–923. doi: 10.1016/j.tiv.2012.04.014. - DOI - PubMed

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