| Abstract | Aflatoksini, kao visoko toksični, mutageni, teratogeni i karcinogeni spojevi zahtijevaju veliku pozornost. Koncentracije aflatoksina u sirovom i obrađenom mlijeku variraju ovisno o geografskom području, klimatskim uvjetima, vlazi i temperaturi. Najčešći aflatoksin u mlijeku je aflatoksin M1 (AFM1) koji je hepatokarcinogeni metabolit dobiven hidroksilacijom aflatoksina B1. U RH najveća dopuštena količina AFM1 u mlijeku i mliječnim proizvodima iznosi 0,05 μg L-1. Za detekciju se koristi ELISA metoda koja zbog svoje jednostavnosti, prihvatljive cijene analize i dostupnosti već dugo čini sastavni dio većih laboratorija unutar različitih institucija na polju medicine, veterine i farmacije. U ovom radu je za određivanje AFM1 u mlijeku validirana metoda LC-MS/MS, koja visoko kotira na polju preciznosti, osjetljivosti, ponovljivosti te linearnosti. Međutim visoka sofisticiranost uređaja, investicijski troškovi te potreban visoko educiran kadar za uspješan rad s uređajem čini ovu metodu manje dostupnom manjim laboratorijima. U postupku validacije su opisani parametri koji utječu na analizu, poput linearnosti, granice detekcije, točnosti, preciznosti i robusnosti. |
| Abstract (english) | Aflatoxins, as highly toxic, mutagenic, teratogenic and carcinogenic compounds, demand big attention.The concentrations of aflatoxins in raw and processed milk vary depending on the geographical area, climate conditions, moisture and temperature. The hepatocarcinogenic metabolite obtained by hidroxylation of aflatoxin B1, aflatoxin M1, is the most common aflatoxin in milk. The highest allowed concentration of AFM1 in milk and dairy products in the Republic of Croatia is 0,05 μg L-1. The method used for detection of AFM1, ELISA method, has been a integral part of laboratories of various institutes in the field of medicine, Animal Health and pharmacy due to its simplicity, acceptable analysis cost and availability. In this thesis, the method validated for the purpose of detection of AFM1 in milk, was the LC-MS/MS method, which quotes highly in the field of precision, sensitivity, repeatability and linearity. However, the high sofistication of the equipment, investment costs and the highly educated staff needed for successful operating with the equipment, makes this method less available for the smaller laboratories. The parametres like linearity, detection limit, accuracy, precision and robustness, which effect the analysis, were described in the validation process. |
