title: Detection of cylindrospermopsin toxin markers in cyanobacterial algal blooms using analytical pyrolysis (Py-GC/MS) and thermally-assisted hydrolysis and methylation (TCh-GC/MS).
abstract: The hepatotoxin cylindrospermopsin (CYN) is produced by freshwater cyanobacteria becoming an emerging threat for human health. Methods for the rapid determination of CYN in environmental samples are needed. Conventional analytical pyrolysis (Py-GC/MS) and thermally-assisted hydrolysis and methylation (TCh-GC/MS) were used to study a CYN standard, two Aphanizomenon ovalisporum cultures (CYN+) and one culture of Cylindrospermopsis raciborskii (CYN-). A micro-furnace pyrolyzer was used directly attached to a GC/MS system fitted with a 30 m × 250 μm × 0.25 μm film thickness column (14% cyanopropyl phenyl, 86% dimethyl polysiloxane pahase composition). Oven temperature was held at 50 °C for 1 min and increased to 100 °C at 30 °C min(-1), from 100 °C to 300 °C at 10 °C min(-1), and stabilized at 300 °C for 10 min using helium (1 mL min(-1)) as carrier gas. Pyrolysis at 500 °C yield over 70 compounds with 20 specific for CYN+ samples. Two peaks containing a diagnostic fragment (m/z 194) were found at 25.0 and 28.9 min only in CYN+ samples. Fewer peaks with limited diagnostic value were released after TCh-GC/MS, including breakdown products and TMAH adducts. A compound was detected that may correspond to the CYN molecule (MW 415 Da) thermoevaporation product after the loss of SO3 (MW 80 Da). This TCh-GC/MS peak (m/z 336) together with the fragments obtained by conventional Py-GC/MS (m/z 194) are diagnostic ions with potential use for the direct detection of CYN toxin in environmental samples at last with an estimated 5 ppm detection threshold.
authors: Área de Toxicología, Facultad de Farmacia, Universidad de Sevilla, C/Profesor García González, 2, 41012 Sevilla, Spain.
authors: V
authors: V
authors: Ríos
authors: Área de Toxicología, Facultad de Farmacia, Universidad de Sevilla, C/Profesor García González, 2, 41012 Sevilla, Spain.
authors: Ana I
authors: AI
authors: Prieto
authors: Área de Toxicología, Facultad de Farmacia, Universidad de Sevilla, C/Profesor García González, 2, 41012 Sevilla, Spain.
authors: Ana M
authors: AM
authors: Cameán
authors: Instituto de Recursos Naturales y Agrobiología de Sevilla, IRNAS-CSIC, Av. Reina Mercedes, 10, 4012 Sevilla, Spain.
authors: F J
authors: FJ
authors: González-Vila
authors: Instituto de Recursos Naturales y Agrobiología de Sevilla, IRNAS-CSIC, Av. Reina Mercedes, 10, 4012 Sevilla, Spain.
authors: J M
authors: JM
authors: de la Rosa
authors: Marine and Environmental Research Centre (CIIMAR/CIMAR), University of Porto, Rua dos Bragas, m289, 4050-123 Porto, Portugal; Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal.
authors: Vitor
authors: V
authors: Vasconcelos
authors: Instituto de Recursos Naturales y Agrobiología de Sevilla, IRNAS-CSIC, Av. Reina Mercedes, 10, 4012 Sevilla, Spain. Electronic address: jag@irnase.csic.es.
authors: J A
authors: JA
authors: González-Pérez
mesh_terms: D000470:Alkaloids; D001427:Bacterial Toxins; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D000458:Cyanobacteria / Q000378:metabolism*; D000087522:Cyanobacteria Toxins; D005068:Eutrophication; D005618:Fresh Water / Q000382:microbiology; D008401:Gas Chromatography-Mass Spectrometry; D006868:Hydrolysis; D008745:Methylation; D013696:Temperature; D014118:Toxins, Biological / Q000032:analysis* / Q000737:chemistry; D014498:Uracil / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000737:chemistry
chemical_list: D000470:Alkaloids; D001427:Bacterial Toxins; D015415:Biomarkers; D000087522:Cyanobacteria Toxins; D014118:Toxins, Biological; D014874:Water Pollutants, Chemical; C089595:cylindrospermopsin; D014498:Uracil
keywords: Analytical pyrolysis; Aphanizomenon ovalisporum; Cylindrospermopsin; Cylindrospermopsis raciborskii; Thermochemolysis; Toxin markers