Alternaria alternata is an important mycotoxin-producing species that occurs on cereals, sunflower seeds, oilseed rape, tomato, carrot, lentils, olives, various fruits, etc. A. alternata produces a number of mycotoxins, including alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), L-tenuazonic acid (TeA), and other less-toxic metabolites. The objective of this study was a toxicological characterization of A. alternata strains isolated from wheat (34 strains) and grapes (11 strains). All the A. alternata strains were cultured on rice to evaluate their toxigenic potential (incubated in the dark at 25°C for 21 days). An instrumental high-performance thinlayer chromatography (HPTLC) for the quantification of ALT, AOH, AME, and TeAin rice cultures has been developed. The mycotoxins were extracted with methanol and ammonium sulfate in water and the extracted solutions were further cleaned up by liquid-liquid extraction. The silica gel layers pre-coated with oxalic acid in methanol were used for separation with toluene-ethyl acetate-formic acid (6:3:1, ν/ν) as the mobil phase. The chromatogram of ALT, AOH, AME was scanned in fluorescence mode after excitation at λ = 254 nm with λ = 400 nm measuring filter: SENS and SPAN parameters were 195 and 15, respectively. The chromatogram for determination of TeA was sprayed with solution of iron trichloride in ethanol to reveal the spots of TeA and scanned in absorbance mode at λ = 254 nm at the SENS and SPAN parameters 145 and 10, respectively. The RF value of ALT under these conditions was 0.25, AOH 0.36, AME 0.49, and TeA 0.30, respectively. The recovery (the mass fraction) was 0.59–0.76 in the range 20–100 mg kg−1 rice cultures. The relative standard deviations of repeatability (RSDr) were 7–19%. The limit of quantification (LOQ) of ALT, AOH, AME was 1.0 mg kg−1, LOQ of TeA was 5.0 mg kg−1 in rice cultures with Alternaria alternata mycelium.
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