A novel biosensor for the detection of zearalenone family mycotoxins in milk

Department of Chemistry and Bioengineering, Tampere University of Technology, P.O. Box, 541, FI- 33101 Tampere, Finland.
Journal of microbiological methods (Impact Factor: 2.03). 11/2009; 80(1):44-8. DOI: 10.1016/j.mimet.2009.10.017
Source: PubMed


In this study, a method for detecting estrogenic mycotoxin residues in milk was developed utilizing bioluminescent whole-cell biosensors. Milk products of various compositions were spiked with the estrogenic mycotoxins zearalenone and its metabolites zearalanone, alpha-zearalanol, beta-zearalanol, alpha-zearalenol and beta-zearalenol. The estrogenic response was detected by a whole-cell biosensor based on a genetically modified Saccharomyces cerevisiae strain that in the presence of an estrogenic compound produces firefly luciferase-enzyme and further light emission within a system provided with D-luciferin substrate. The results show that the yeast sensor reacts to mycotoxins with typical sigmoidal response at nanomolar concentrations. The response differs in different milk products with regard to the fat content of the milk. Due to short assay time of less than 3h and automation the approach can be used as a bioavailability and activity screening method prior to more detailed chemical analysis.

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    • "DON is mostly biotransformed within the rumen and detected in milk as DON and deepoxy-deoxynivalenol (DOM-1) [20–22], whereas T-2 has been detected in the milk without metabolization [23] and metabolized into HT-2, NEO, 4-deacetylneosolaniol, and 4 more unknown metabolites [24]. Metabolites could have different toxic effects than the parent mycotoxin [25] [26]. More studies are needed in order to increase knowledge regarding this aspect. "
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    ABSTRACT: An LC-MS/MS (QqQ) method has been developed and validated for simultaneous determination of the following trichothecenes in UHT cow milk: nivalenol (NIV), deoxynivalenol (DON), deepoxy-deoxynivalenol (DOM-1), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), neosolaniol (NEO), diacetoxyscirpenol (DAS), fusarenon X (FUS-X), T-2 and HT-2 toxins. Sample treatment is simple and based on the extraction with acetonitrile (ACN), acidified with 0.2% formic acid, followed by a purification process, adding sodium acetate to the ACN/water extract in order to separate aqueous phase and, consequently, polar components of the milk. Validation of the method for all the 10 mycotoxins was successful; validation parameters taken into account were as follows: limits of detection (LOD) and quantification (LOQ), linearity, precision (within-day and between-day variability), recovery, matrix effect and stability. The LODs were 10.1, 2.5, 1.5, 1.9, 0.1, 0.5, 1.0, 0.08, 0.4 and 0.05ng/mL for NIV, DON, DOM-1, FUS-X, NEO, 3-ADON, 15-ADON, DAS, HT-2 and T-2, respectively. Mean recovery values (obtained in intermediate precision conditions) were between 63.5 and 75.8 (RSDR≤15%) for all the mycotoxins. All the mycotoxins suffered from matrix effects, especially DON.
    Journal of Chromatography A 09/2015; DOI:10.1016/j.chroma.2015.09.069 · 4.17 Impact Factor
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    • "After addition of buffered sample, a platinum electrode was employed as reference electrode. Vaelimaa et al. (2010) developed a bioluminescent whole-cell biosensor for the detection of ZEA, α-ZOL, β-ZOL, ZAN, α-ZAL and β-ZAL in milk products. A genetically modified Saccharomyces cerevisiae strain was used as detection system. "
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    ABSTRACT: This review highlights developments in mycotoxin analysis and sampling over a period between mid-2009 and mid-2010. It covers the major mycotoxins aflatoxins, Alternaria toxins, ergot alkaloids, fumonisins, ochratoxin, patulin, trichothecenes, and zearalenone. New and improved methods for mycotoxins continue to be published. Immunological-based method developments continue to be of wide interest in a broad range of formats. Multimycotoxin determination by LC-MS/MS is now being targeted at the specific ranges of mycotoxins and matrices of interest or concern to the individual laboratory. Although falling outside the main emphasis of the review, some aspects of natural occurrence have been mentioned, especially if linked to novel method developments
    World Mycotoxin Journal 02/2011; 4(1). DOI:10.3920/WMJ2010.1249 · 2.16 Impact Factor
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    ABSTRACT: Mycotoxins are secondary metabolites produced by several filamentous fungi that grow on various food and feed. These compounds elicit a wide spectrum of toxicological effects, representing a health risk for both, humans and animals. Mycotoxin analysis is an important tool in controlling fungal contamination of food and feed. This article reviews the techniques used for their determination, and the advantages and limitations of each method are critically discussed.
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