Simultaneous Determination of Deoxynivalenol, Deoxynivalenol-3-Glucoside and Nivalenol in Wheat Grains by HPLC-PDA with Immunoaffinity Column Cleanup

Abstract

Deoxynivalenol-3-glucoside (D3G) is a modified mycotoxin formed by the metabolism of plants through the conjugation of deoxynivalenol (DON) with glucose. Toxicology studies of D3G for human and animal health are still under investigation, and the development of practical and reliable methods for its direct determination, especially in cereal matrices, is of great importance. In the present study, a methodology for simultaneous determination of D3G, DON, and nivalenol (NIV) in wheat grains, using immunoaffinity column (IAC) cleanup, separation by C18 column and detection by ultraviolet (UV) absorption, was optimized and inhouse validated. The results demonstrated adequate values of D3G recovery from IAC and spiked samples. Intraday precision, linearity, limit of detection and limit of quantification (LOQ) were also adequate for the determination of these mycotoxins. Range of applicability varied from 47.1 to 1000 μg/ kg for D3G and from 31.3 to 1000 μg/kg for DON and NIV, with recovery ranging from 84.7 ± 7.2 % to 112.3 ± 8.1 %. A high incidence of D3G (41.2 %, all samples <LOQ) was verified in commercial samples of wheat grains and whole wheatflour (n = 17). Also, the presence of D3G occurred simultaneously with DON in 100 % of the D3G-positive samples. DON levels varied from <LOQ to 325.8 μg/kg, and NIV was detected in only 29.5 % (from <LOQ to 140.6 μg/kg). To the best of our knowledge, this is the first method of simultaneous determination of NIV, D3G, and DON by highperformance liquid chromatography with photodiode array detector (HPLC-PDA) reported until now.

Natural Occurrence of Nivalenol, Deoxynivalenol, and Deoxynivalenol-3-Glucoside in Polish Winter Wheat

“…Therefore they enabled simultaneous analyses of NIV, DON, and DON-3G. Advantages of the columns have recently been successfully utilized also by other authors (Trombete et al [ 25 ], Geng et al [ 26 ], Yoshinari et al [ 27 ]).…”

“…DON-3G was found in 44–78% of samples (depending on the cultivation region) at concentrations ranging from 40 to 356 µg/kg. Trombete et al studied 17 samples of wheat cultivated in Portugal [ 25 ]. NIV was found in 29.5% of the samples at concentrations ranging from 31.3 to 140.6 µg/kg.…”

Mycotoxin Biomarkers of Exposure: A Comprehensive Review

“…NIV is chemically classified as a type B trichothecene. NIV is a field‐contaminant, and has been observed in wheat (Trombete et al., ). NIV has demonstrated immunotoxicity, hematotoxicity, myelotoxicity, and developmental and reproductive toxicity.…”

Contents variation analysis of free amino acids, nucleosides and nucleobases in semen sojae praeparatum fermentation using UFLC‐QTRAP MS

“…The method was fully validated in accordance with guidelines on linearity, precision, recovery, detection limit, quantification limit and stability (Trombete et al, ). Calibration curves were generated by plotting peak area against the concentration of standard solutions.…”