Identification of type B trichothecenes and zearalenone in Chilean cereals by planar chromatography coupled to mass spectroscopy

ABSTRACT

High-performance thin-layer chromatography (HPTLC) and HPTLC coupled with mass spectrometry (MS) methods were described for the simultaneous determination of zearalenone (ZEA); type B trichothecenes (TCT-B); nivalenol (NIV) and deoxynivalenol (DON) along with its acetylated derivatives: 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON). The extract samples were cleaned-up with Bond Elut Mycotoxin^® solid-phase extraction cartridges. Then, separation was performed on HPTLC silica gel 60 F₂₅₄ plates using toluene, ethyl acetate and formic acid (1:8:1 v/v/v) as mobile phase. Derivatisation was then performed with 10% aluminium trichloride in 50% methanol. Mycotoxin standards and spiked cereals grains were identified by UV spots at 366 nm, with retention factors (R_F) of 0.20 (NIV), 0.39 (DON), 0.45 (15-ADON), 0.50 (3-ADON) and 0.60 (ZEA). Some parameters of validation were determined. Calibration data (n = 5) fitted a linear regression model with determination coefficients, R² > 0.99. The recovery was determined in triplicate at two levels, ranging from 84.3 ± 2.2% to 114.2 ± 11.7%. Detection limits ranged from 80 to 120 μg kg⁻¹ and quantification limits ranged from 120.0 to 200 μg kg⁻¹. The analysis by HPTLC/electrospray (ESI)-MS in negative mode confirmed the presence of TCT-B and ZEA standards in Chilean cereals with mass signals at m/z 355, 371, 337, and 317 for DON, NIV, 3-ADON and 15-ADON, and ZEA, respectively.

KEYWORDS:

• Cereal
• high-performance thin-layer chromatography
• mass spectrometry
• mycotoxin

Acknowledgments

This work is part of Diego Jorquera’s thesis to obtain the degree of Master in Pharmaceutical Sciences from the University of Concepcion, Chile. The authors wish to thank the Interdisciplinary Research Laboratory in Mycotoxins (LIM), Department of Food Sciences and Technology, Faculty of Pharmacy, University of Concepcion.

Disclosure statement

No potential conflict of interest was reported by the author(s).