## 1 Introduction  
T-2 toxin (T-2) and HT-2 toxin (HT-2) belong to the group of contaminants known as trichothecenes, which are part of the largest group of _Fusarium_ mycotoxins, comprising more than 150 compounds. The basic structure of all trichothecenes is a tetracyclic sesquiterpene with a spiro-epoxide group at C-12 and C-13, as well as an olefinic double bond between C-9 and C-10. According to the substituents of the tetracyclic ring system, trichothecenes are grouped into four different types (A-D). T-2 and HT-2 belong to the type A trichothecenes, which are characterized by an esterified or free hydroxyl group at C-8 or an unsubstituted C-8. The structures of T-2 and HT-2 toxins differ only in one functional group; the T-2 toxin is acetylated at C-4, while the HT-2 toxin is not acetylated. The T-2 toxin is readily converted to HT-2 by metabolism without losing its toxicity; therefore, T-2 and HT-2 are usually evaluated together [1].  
In 2011, the European Food Safety Authority (EFSA) published a scientific opinion on the risks to human and animal health related to the presence of the T-2 and HT-2 toxins in food and feed [2]. In this opinion, the EFSA Panel on Contaminants in the Food Chain concluded that T-2 is a carcinogen, mutagen, and immunosuppressant. Thus far, no legal limits have been defined in Europe for T-2/HT-2 mycotoxins, and only a recommendation is available (Recommendation 2013/165/EU; Table 1). However, the recommendation specifically asks the member states to monitor for the presence of both T-2 and HT-2 in food and feed. This means that, in the near future, the laboratories in charge of food inspections will need fast and reliable analytical tools for analyzing a large number of samples in a short time.  
The main analytical methods used for the T-2 and HT-2 toxin determination in cereal grains are chromatographic methods--mainly gas chromatography coupled with flame ionization detection (FID), electron capture detection (ECD), and mass spectrometry (MS) detection [3; 4; 5]. Additionally, liquid chromatography coupled with spectrofluorometric detection or mass spectrometry has been indicated for T-2 and HT-2 analysis [5; 6; 7; 8; 9]. [PERSON] et al. [10] published a sensitive HPLC method for the analysis of T-2 toxin in cereals. This method includes an immunoaffinity column (IAC) clean-up after extraction and a derivatization step with 1-anthroylnitrile and 4-dimethylaminopyridine before fluorescence detection. Nevertheless, the ELISA approach is more employed due to its simplicity and rapidity [5; 11]. However, many authors have reported some selectivity limitations of these approaches [12; 13].  
Regarding the performances of analytical methods for the research of drug residues and environmental contaminants in food, European legislation requires the validation of both screening and confirmatory methods, according to the European Decision No. 2002/657/EC [14]. This decision defines a list of the definitions and requirements to fulfill through the validation procedure.  
The aim of our work was to validate and then compare the ELISA, as a screening method, and ultra performance liquid chromatography with fluorescence detection (UPLC/FLD), as a confirmatory method, for the quantitative determination of the T-2 and HT-2 toxins in cereals. The comparison was carried out while considering both the obtained validation parameters (selectivity, precision, recovery, robustness, and measurement uncertainty) and the results coming from the analysis of real samples.