Validation of a multimycotoxin LC-MS/MS method

Zadatak ovog rada bio je validirati metodu za istovremenu analizu devet mikotoksina korištenjem LC-MS/MS-a. Validacijom ove metode ispitivana je sposobnost metode za određivanje trihotecenskih mikotoksina u uzorcima pšenice. Spektrometar masa je podešen za rad u pozitivnom i negativnom ionizacijskom modu, te je izvršena optimizacija ostalih parametara (napon za lokaliziranje i razbijanje, ulazni napon, kolizijska energija kao i izlazni potencijal kolizijske ćelije). Prije analize obavljena je kalibracija otopinama standarda, te je dokazana prihvatljiva linearnost metode. Uzorak pšenice podvrgnut je ekstrakciji i pročišćavanju na SPE koloni, prije analize na LC-MS/MS-u. Osim linearnosti, ispitivani validacijski parametri su bili repetabilnost, reproducibilnost, limit detekcije (LOD), limit kvantifikacije (LOQ) i iskorištenje. Obzirom na navedene parametre, metoda je zadovoljila zakonsku regulativu, osim vrijednosti iskorištenja za nivalenol, fuzarenon X, T-2 i HT-2 toksin, koje je bilo ispod propisanog minimuma.The aim of this work was to validate a multimycotoxin LC-MS/MS method for simultaneous analysis of nine mycotoxins. The method’s ability to measure trichothechene mycotoxins in wheat was estimated. The mass spectrometer was operated in both positive and negative ionization mode. Other parameters, including declustering potential, focusing potential, entrance potential, collision energy and collision cell exit potential, were optimized as well. Calibration was performed using standard solutions standard and acceptable linearity was obtained. Wheat sample extracts were subjected to SPE column clean-up prior to LC-MS/MS analysis. In addition to linearity, tested validation parameters were also repetability, reproducibility, detection limit (LOD), quantitation limit (LOQ) and recovery. According to the results, this method met regulatory demands, except for nivalenol, fusarenon X, T-2 and HT-2. Recoveries of these toxins were below acceptable ranges