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HPLC-Based Aflatoxin Detection Protocol

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Yeast extract, potato dextrose agar and sucrose were purchased from Biolife (Italy) while aflatoxin standard mix (B1, G1, B2, G2) was purchased from Biopure (Austria). Acetonitrile and methanol (both HPLC grade) were obtained from Merck (Germany). Ammonium acetate and glacial acetic acid (p.a.) were purchased from Sigma Aldrich (Vienna, Austria). A Purelab Ultra system (ELGA LabWater, Celle, Germany) was used for ultrapure water preparation. Standards of A. flavus metabolites were purchased from Various research groups or from the following commercial sources: Romer Labs®Inc.(Tulln, Austria), Sigma–Aldrich (Vienna, Austria), Iris Biotech GmbH (Marktredwitz, Germany), Axxora Europe (Lausanne, Switzerland) And LGC Promochem GmbH (Wesel, Germany) and prepared as described by Malachova et al.46 (link).
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2

Antioxidant Capacity Evaluation Protocol

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Yeast extract, potato dextrose agar, agar, and sucrose were purchased from Biolife (Milan, Italy). Ethylenediaminetetraacetic acid disodium salt (EDTA-2Na) was from Pharmacia Biotech (Uppsala, Sweden), stabilized 3% solution of hydrogen peroxide (H2O2) was obtained from Fluka (Buchs, Switzerland), and potassium cyanide was purchased from Sigma Aldrich (Taufkirchen, Germany). Hydrochloric acid as purchased from Merck (Darmstadt, Germany). Trichloroacetic acid (TCA) and ascorbic acid were from Kemika (Zagreb, Croatia), absolute ethanol was from Panreac (Barcelona, Spain), while butylated hydroxytoluene (BHT) and 2-thiobarbituric acid (TBA) was obtained from Acros Organics (Morris Plains, New Jersey, USA). Superoxide dismutase from bovine erythrocytes (3000 U mg −1 protein) (SOD) and xanthine oxidase from bovine milk (0.4-1.0 U mg −1 protein) (XOD) were purchased from Sigma-Aldrich (Taufkirchen, Germany). Formic acid was purchased from Fluka (Buchs, Switzerland). Aflatoxin standard mix (B1, B2, G1, G2) was purchased from Biopure (Tulln, Austria). Acetonitrile and methanol (both HPLC grade) were purchased from J. T. Baker (Radnor, PA, USA).
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3

Preparation of Mycotoxin Standards

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Yeast extract, potato dextrose agar, malt extract agar and sucrose were purchased from Biolife (Italy). AFT standard mix (B1, G1, B2, G2) was purchased from Biopure (Austria). Acetonitrile and methanol (HPLC grade both) were obtained from Merck (Germany). Ammonium acetate and glacial acetic acid (p.a.) were purchased from Sigma Aldrich (Vienna, Austria). For ultrapure water preparation, a Purelab Ultra system (ELGA LabWater, Celle, Germany) was used. Standards of A. flavus metabolites were collected from various research groups or purchased from the following commercial sources: Romer Labs®Inc.(Tulln, Austria), Sigma–Aldrich (Vienna, Austria), Iris Biotech GmbH (Marktredwitz, Germany), Axxora Europe (Lausanne, Switzerland) And LGC Promochem GmbH (Wesel, Germany). Purchased standards were prepared according to Malachova et al.50 (link).
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