Analysis of mycotoxins in barley using ultra high liquid chromatography high resolution mass spectrometry: Comparison of efficiency and efficacy of different extraction procedures Josep Rubert a,n , Zbynek Dzuman b , Marta Vaclavikova b , Milena Zachariasova b , Carla Soler a , Jana Hajslova b a Departament de Medicina Preventiva, Facultat de Farm acia, Universitat de Valencia, Av. Vicent Andr  es Estell  es s/n, 46100 Burjassot (Val  encia), Spain b Department of Food Chemistry and Analysis, Faculty of Food and Biochemical Technology, Institute of Chemical Technology, Technicka ´ 5, 166 28 Prague 6, Prague, Czech Republic article info Article history: Received 6 March 2012 Received in revised form 26 June 2012 Accepted 5 July 2012 Available online 20 July 2012 Keywords: QuEChERS MSPD Clean-up Solid–liquid extraction Mycotoxins Orbitrap abstract The effectiveness of four extraction methods (modified QuEChERS, matrix solid-phase dispersion (MSPD), solid–liquid extraction (SLE) and solid-phase extraction (SPE) clean-up) were evaluated for simultaneous determination of 32 mycotoxins produced by the genus Fusarium, Claviceps, Aspergillus, Penicillium and Alternaria in barley by ultra high pressure liquid chromatography coupled to ultra-high resolution mass spectrometry (UHPLC-Orbitrap s MS). The efficiency and efficacy of extraction methods were evaluated and compared in number of extracted mycotoxins and obtained recoveries. From the one point of view, QuEChERS procedure was fast and easy, as well as it was able to successfully extract all selected mycotoxins. On the other hand, SLE method, MSPD and SPE clean-up method did not extract adequately all selected mycotoxins and recoveries were not suitable enough. Thereby, method employing QuEChERS extraction connected with UHPLC-Orbitrap s MS was developed to quantify 32 mycotoxins in barley within this study. Analytical method was validated and recoveries ranged from 72% to 101% for selected mycotoxins with only one exception nivalenol (NIV) and deoxynivalenol-3- glucoside (D3G), which were lower than 67%. Relative standard deviations (RSD) were lower than 17.4% for all target mycotoxins. The lowest calibration levels (LCLs) ranged from 1 to 100 mg/ kg. Validated method was finally used for monitoring mycotoxins in a total of 15 Czech barley samples, when only Fusarium toxins representatives were detected in 53% of samples and the mycotoxins with the highest incidence were enniatins. & 2012 Elsevier B.V. All rights reserved. 1. Introduction Cultivated for over 10,000 years, barley is one of the oldest domesticated grain crops. There are different varieties of barley, which have been developed during a long time. Actually barley is the world’s fourth most important crop and an important staple in many countries. The largest commercial producers of barley are Canada, United States, Russia, Germany, France and Spain [1,2]. Moreover, countries like Czech Republic, barley is a crop with a great economical importance, with a cultivation area of about 400,000 ha [3]. The use of barley is predominantly focused on the production of malt by malting process intended for beer produc- tion. Even though, the malt is also used for the manufacture of distilled spirits, such as whisky, as well as sirups, coffee substitutes, and some other cereal-based foods. Moreover, malt or barley derivatives are used for feed production. Thereby, this product is commonly consuming by humans and animals [4]. Normally, the plant of barley can easily grow in different climatic regions. Unfortunately, this particular capacity does vulnerable to be colonised by various toxinogenic fungi, some of them can be able to produce mycotoxins [5,6]. These toxins can cause both acute and chronic effects for humans and animals [7,8]. For this reason, well-known mycotox- ins, such as aflatoxins (AFs), ochratoxin A (OTA) and some Fusarium toxins have been classified by International Agency for Research on Cancer (IARC) and regulated by European Union [9–11]. On the other hand, there are other mycotoxins, such as enniatins, beauvaricin or ergot alkaloids, which have not been classified nor legislated up to now. The starting point of the monitoring of mycotoxins began to be focused on legislated mycotoxins [12,13], but step-by-step the range was also extended to emerging mycotoxins. In fact, several recent works have been focused only on these new and emerging mycotoxins [14–16]. Contents lists available at SciVerse ScienceDirect journal homepage: www.elsevier.com/locate/talanta Talanta 0039-9140/$ - see front matter & 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.talanta.2012.07.010 n Correspondence author. Tel.: þ34 96 3543091; fax: þ34 96 3544954. E-mail address: josep.rubert@uv.es (J. Rubert). Talanta 99 (2012) 712–719