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      New Method for Simultaneous Determination of Microcystins and Cylindrospermopsin in Vegetable Matrices by SPE-UPLC-MS/MS

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          Abstract

          Cyanotoxins are a large group of noxious metabolites with different chemical structure and mechanisms of action, with a worldwide distribution, producing effects in animals, humans, and crop plants. When cyanotoxin-contaminated waters are used for the irrigation of edible vegetables, humans can be in contact with these toxins through the food chain. In this work, a method for the simultaneous detection of Microcystin-LR (MC-LR), Microcystin-RR (MC-RR), Microcystin-YR (MC-YR), and Cylindrospermopsin (CYN) in lettuce has been optimized and validated, using a dual solid phase extraction (SPE) system for toxin extraction and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Results showed linear ranges (5–50 ng g −1 f.w.), low values for limit of detection (LOD) (0.06–0.42 ng g −1 f.w.), and limit of quantification (LOQ) (0.16–0.91 ng g −1 f.w.), acceptable recoveries (41–93%), and %RSD IP values for the four toxins. The method proved to be robust for the three variables tested. Finally, it was successfully applied to detect these cyanotoxins in edible vegetables exposed to cyanobacterial extracts under laboratory conditions, and it could be useful for monitoring these toxins in edible vegetables for better exposure estimation in terms of risk assessment.

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          Most cited references53

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          Trends in quality in the analytical laboratory. II. Analytical method validation and quality assurance

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            Cylindrospermopsin: a potent hepatotoxin from the blue-green alga Cylindrospermopsis raciborskii

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              Oral toxicity of the cyanobacterial toxin cylindrospermopsin in male Swiss albino mice: determination of no observed adverse effect level for deriving a drinking water guideline value.

              The cyanobacterial toxin cylindrospermopsin (CYN) is a frequent contaminant of freshwaters throughout the world, including those that are sources of drinking water. The first cases of human poisoning attributed to this toxin occurred from a treated drinking water supply in Queensland, Australia, in 1979. The toxin causes extensive damage to the liver, kidneys, spleen, heart, and other organs. It is known to be a potent protein synthesis inhibitor, but there is mounting evidence for genotoxicity and that it metabolizes to even more toxic forms. As part of a risk assessment process leading to a guideline for a safe drinking water level for this toxin, we performed a series of experiments to determine a no-observed-adverse-effect level (NOAEL) for this toxin. In the first trial male mice were exposed to CYN-containing cyanobacterial extract in their drinking water (0-657 microg CYN kg(-1) day(-1)) for 10 weeks. In the second trial mice received purified CYN by daily gavage (0-240 microg CYN kg(-1) day(-1)) for 11 weeks. Body and organ weights were recorded; urine, serum, and hematology analyses were performed; and histopathological examination of tissues was carried out. Body weights were significantly increased at low doses (30 and 60 microg kg(-1) day(-1)) and decreased at high doses (432 and 657 microg kg(-1) day(-1)). Liver and kidney weights were significantly increased at doses of 240 microg kg(-1) day(-1) and 60 microg kg(-1) day(-1), respectively. Serum bilirubin levels were significantly increased and bile acids significantly decreased at doses of 216 microg kg day(-1) and greater. Urine total protein was significantly decreased at doses above 60 microg kg(-1) day(-1). The kidney appeared to be the more sensitive organ to this toxin. If it is assumed that increased organ weights and changes in functional capacity are responses to an underlying toxic effect, then the NOAEL based on this data is 30 microg kg(-1) day(-1), which, with standard calculations and uncertainty factors, provides a proposed guideline safety value of 1 microg/L in drinking water. Copyright 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 94-103, 2003.
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                Author and article information

                Journal
                Toxins (Basel)
                Toxins (Basel)
                toxins
                Toxins
                MDPI
                2072-6651
                08 October 2018
                October 2018
                : 10
                : 10
                : 406
                Affiliations
                [1 ]Area of Toxicology, Faculty of Pharmacy, University of Sevilla, C/Profesor García González 2, 41012 Sevilla, Spain; ldiezquijada@ 123456us.es (L.D.-Q.); anaprieto@ 123456us.es (A.I.P.O.); mllana@ 123456us.es (M.L.-R.-C.); angelesjos@ 123456us.es (A.J.); camean@ 123456us.es (A.M.C.)
                [2 ]CIIMAR/CIMAR—Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Terminal de Cruzeiros do Porto de leixões, Av General Norton de Matos, 4450-208 Matosinhos, Portugal; amoclclix@ 123456gmail.com (A.C.); vmvascon@ 123456fc.up.pt (V.V.)
                [3 ]Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal
                Author notes
                [* ]Correspondence: rguzman1@ 123456us.es ; Tel.: +34-954-556-762
                Author information
                https://orcid.org/0000-0001-6714-4461
                https://orcid.org/0000-0003-3585-2417
                https://orcid.org/0000-0003-1524-748X
                Article
                toxins-10-00406
                10.3390/toxins10100406
                6215191
                30297653
                bb582817-ba10-4b09-8519-54759159db0a
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 07 September 2018
                : 06 October 2018
                Categories
                Article

                Molecular medicine
                microcystins,cylindrospermopsin,method validation,uplc-ms/ms,lettuce
                Molecular medicine
                microcystins, cylindrospermopsin, method validation, uplc-ms/ms, lettuce

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