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. 2024 May 13;13(10):1501.
doi: 10.3390/foods13101501.

Identification of Biotransformation Products of T-2 Toxin in HepG2 Cells Using LC-Q-TOF MS

Mercedes Taroncher  1   2 , Veronica Zingales  1   2 , Yelko Rodríguez-Carrasco  1   2 , María José Ruiz  1   2
Affiliations

Identification of Biotransformation Products of T-2 Toxin in HepG2 Cells Using LC-Q-TOF MS

Mercedes Taroncher et al. Foods. .

Abstract

The T-2 toxin (T-2) is a type A trichothecene found in cereals. The formation of metabolites is a frequent cause of mycotoxin-induced toxicity. In this work, the conversion of T-2 during biotransformation reactions in HepG2 cells was evaluated. For this, HepG2 cells were exposed to 30 (IC50/2) and 60 (IC50) nM of T-2 for 0, 1, 2, 3, 6, 8 and 24 h, and the concentrations of T-2 and its metabolites HT-2, T2-triol, T2-tetraol and neosolaniol were determined in both the cell fraction and culture medium through liquid chromatography coupled to high-resolution mass spectrometry-time of flight (LC-Q-TOF MS). Results showed a fast metabolization of T-2 (>90%) during the first 2 h, with HT-2 as its main (>95%) biotransformation product. The cell fraction showed higher levels (p < 0.05) of HT-2 (39.9 ± 2.1 nM) compared to the culture medium (12.53 ± 2.4 nM). This trend was also observed for the identified metabolites. T2-triol reached its maximum concentration (1.7 ± 0.4 nM) at 2 h, and at later times a time-dependent increase in the T2-tetraol and neosolaniol concentrations was observed. The identification of T-2 metabolites shows the need to continue combined toxicity studies of mycotoxins for a correct risk characterization of these natural contaminants.

Keywords: HepG2; LC-Q-TOF MS; T-2; biotransformation; in vitro.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Concentrations of T-2, HT-2, T2-triol, T2-tetraol and Neo identified and quantified by LC-Q-TOF MS in the cell fraction of HepG2 cells after 0, 1, 2, 3, 6, 8 and 24 h of 60 nM of T-2 exposure. Values are expressed as mean ± SEM (n = 3). Values with different superscript letters for each metabolite are significantly different (p ≤ 0.05). The black superscript letter indicates the statistic of T-2. The bold superscript letter indicates the statistic of HT-2. The italic superscript letter indicates the statistic of T2-triol. The bold italic superscript letter indicates the statistic of T2-tetraol. The underlined superscript letter indicates the statistic of Neo.
Figure 2
Figure 2
Concentrations of T-2, HT-2, T2-triol, T2-tetraol and Neo identified and quantified by LC-Q-TOF MS in the culture medium of HepG2 cells after 0, 1, 2, 3, 6, 8 and 24 h of 60 nM of T-2 exposure. Values are expressed as mean ± SEM (n = 3). Values with different superscript letters for each metabolite are significantly different (p ≤ 0.05). The black superscript letter indicates the statistic of T-2. The bold superscript letter indicates the statistic of HT-2. The italic superscript letter indicates the statistic of T2-triol. The bold italic superscript letter indicates the statistic of T2-tetraol. The underlined superscript letter indicates the statistic of Neo.
Figure 3
Figure 3
Mass spectra of T-2 standard. Precursor ion at m/z 484.2541 ([H + NH4]+) and major product ions at m/z 365.2674 and m/z 205.2258.
Figure 4
Figure 4
Total ion chromatogram (m/z 383.2064 [M + H]+) and mass spectra of T2-triol standard. The major product ion peak was obtained at m/z 303.2142.
Figure 5
Figure 5
Total ion chromatogram (m/z 299.1495 [M + H]+) and mass spectra of T2-tretraol standard. The major product ion peak was obtained at m/z 285.4261.
Figure 6
Figure 6
Total ion chromatogram (m/z 400.1966 [M + NH4]+) and mass spectra of Neo standard. The major product ion peak was obtained at m/z 245.2163.
Figure 7
Figure 7
Total ion chromatogram (m/z 442.2435 [M + NH4]+) and mass spectra of HT-2 standard. The major product ion peak was obtained at m/z 263.1211.
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