An LC-IT-MS/MS-Based Method to Determine Trichothecenes in Grain Products
Marcin Brya
Renata Jdrzejczak
Krystyna Szymczyk
Marek Roszko
Mieczysaw W. Obiedziski
The aim of this work was to evaluate the usefulness of the ion trap mass spectrometry coupled to high-performance liquid chromatography for simultaneous determination of selected trichothecenes (nivalenol, deoxynivalenol, fusarenon-X, neosolaniol, 3-acetyl-deoxynivalenol, diacetoxyscirpenol, HT2 and T-2 toxins) in grain products. These compounds were extracted from the grain products and then cleaned up with the developed, simple and robust procedure using some mixture of neutral alumina, charcoal and diatomaceous earth. Method recovery was 88-125 % depending on combination of the analysed mycotoxins, sample matrix and the fortification level. Method precision expressed by relative standard deviation ranged from 2.6 to 27.4 %. The concentrations of the selected trichothecenes have been determined in 94 samples of cerealbased products. Maize-based next to wheat-based products were the most contaminated with deoxynivalenol, neosolaniol, 3-acetyl-deoxynivalenol, diacetoxyscirpenol and HT-2 toxin. In 83 % of wheat-based products, deoxynivalenol was determined at the average level of 249 g kg1. The highest concentration of deoxynivalenol2,026 g kg1 (476 471 g kg1 on the average)was found in the maize-based product. Other mycotoxins were found much less frequently: 3-acetyl-deoxynivalenol in only one sample at the concentration of 59 g kg1, neosolaniol, HT-2 toxin and diacetoxyscirpenol in a few samples on average concentrations close to respective limits of quantification.
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Trichothecenes are mycotoxins produced by various Fusarium
fungi such as Fusarium sporotrichioides (the T-2 toxin and
other trichothecenes), Fusarium graminearum , Fusarium
culmorum and Fusarium crookwellense (deoxynivalenol
(DON), nivalenol (NIV), fusarenon (FUS-X)). They are the
most often encountered mycotoxins in cereal grains. Many hot
spots of contamination with Fusarium- produced mycotoxins in
wheat and maize grains have been located in China, India and
USA (Charlton and Holstege 2010).
The trichothecin toxin compound itself was for the first
time isolated from Trichothecium roseum and described by
Freeman and Morrison (1948). Later isolated trichothecenes
included diacetoxyscirpenol (DAS), the T-2 toxin, NIV and
DON. More than 180 various trichothecenes and their
derivatives have been characterized so far from which the T-2,
HT2 toxins and NIV exhibit the largest toxicity among all of them
(Yazar and Omurtag 2008).
Trichothecenes are multi-ring sesquiterpenes with a
common 12,13-epoxytrichothecene ring (Krska et al. 2007; Yazar
and Omurtag 2008; Monaci et al. 2011; Mateo et al. 2002).
Because of some subtle but significant differences in basic
chemical structures, they have been classified into four types
referred to as types A, B, C and D trichothecenes, of which the
first two are the most important ones (Berthiller et al. 2005;
Charlton and Holstege 2010). Each type B trichothecene has at
its C-8 position the carbonyl group, whilst the group is absent in
type A trichothecenes. Type A trichothecenes include such
compounds as scirpentriol, 15-monoacetoxyscirpenol, DAS,
HT-2 and T-2 toxins, T-2 triol, T-2 tetraol and neosolaniol
(NEO). Type B trichothecenes include such compounds as
DON, 3- and 15-acetyl-deoxynivalenol (3- and 15-ADON),
NIV and FUS-X. Basic chemical structure and functional
groups of the most important trichothecenes are shown in
Fig. 1 (Schollenberger et al. 2008).
DON is the most often encountered and studied
trichothecene toxin. It is produced mainly by F. graminearum and F.
culmorum (DallAsta et al. 2010). Even if DON is not
particularly toxic, its low-to-moderate doses constantly find their
way into human/animal organisms via food and feed chain
(Mankeviciene 2010). Most severe problems brought about
the toxin appear in animal breeding (Lauber et al. 2001;
Mankeviciene 2010).
The T-2 toxin is another mycotoxin frequently found in
foodstuffs. It strongly inhibits the synthesis of DNA and RNA
(Medina and Magan 2011), influences immunologic system,
exhibits cytotoxicity (Lattanzio et al. 2008; Busman et al.
2011) and inhibits synthesis of proteins both in vivo and
in vitro (Ingle et al. 2010; Medina and Magan 2011).
Inhaled T-2 toxin was 10 times more toxic than T-2 toxin
taken orally (Schwake-Anduschus et al. 2010; Ingle et al.
2010). Microflora present in gastrointestinal tract of every
mammal converts T-2 toxin into several metabolites, mainly
into the HT-2 toxin that is easily absorbed into the blood
(Medina and Magan 2011). Therefore, toxic effects caused
in vivo by HT-2 toxin are partly included in the evaluation of
toxicity of T-2 toxin (Lattanzio et al. 2008; Busman et al.
2011; Medina and Magan 2011).
European Commissions Scientific Committee on Food
(SCF) has specified tolerable daily intake (TDI) regarding
some Fusarium mycotoxins, including TDI for DON, NIV
and HT-2 and T-2 toxins equal to 1, 0.7 and 0.06 g/k (...truncated)