Validation and implementation of validated methods for quantification of aflatoxin in nuts

Cilj rada bio je prikazati kvalitetu ispitivanja uzoraka na prisutnost aflatoksina u orašastim polodovima kroz parametre validacije, te dokazati efikasnost ELISA tehnike kao pouzdane metode u kvantificiranju velikog broja uzorka. Ispitani uzorci (orašasti plodovi) analizirani su na komercijalno dostupnom kitu RomerLabsa ("Total Aflatoxin Assay") u rasponu koncentracija od 1 do 20 µg/kg koji sadrži sve potrebne reagense i standarde za analizu aflatoksina. Rezultati su bili statistički obrađeni kroz srednju vrijednost, standardnu i relativno standardnu devijaciju. Za određivanje koncentracije aflatoksina u orašastim plodovima ELISA testom provjeravali su se slijedeći validacijski parametri: lineranost, preciznost, iskorištenje, stabilnost mjernih otopina, granica detekcije i granica kvantifikacije, a sve u cilju osiguranja kvalitete dobivenih rezultata. U slučaju kontaminiranih uzorka ili pozitivnih rezultata uzorci se dodatno kontroliraju potvrdnom tehnikom tekućinske kromatografije vezanog sustava sa dvostrukom masenom spektrofotometrijom LC-MS/MS.Provedbom validacije metode za utvrđivanje aflatoksina u orašastim plodovima ELISA tehnikom stavila se osnova za kontinuirano praćenje aflatoksina u analiziranoj vrsti hrane. Laboratorij ima mogućnosti da na brzi i pouzdan način utvrdi da li se radi o kontaminiranoj hrani što je neophodno kada se radi o uvozu takve hrane u RH.The purpose of this final paper was to show the quality of test samples for the presence of aflatoxins in tree nuts trought validation parameters and prove of effectiveness of ELISA tehnique as reliable method of quantifying a larage number of samples.Tested samples (nuts) were analyzed on a commercially available kit RomerLabsa ("Total Aflatoxin Assay") in the concentration range 1-20 mg / kg, which contains all necessary reagents and standards for analysis of aflatoxin. The results were statistically processed through mean value, standard and relative standard deviation. To determine the concentration of aflatoxin in tree nuts ELISA test to verify the following validation parameters are: lineranost, accuracy, recovery, limits of detection and quantification, all in order to ensure quality results. In the case of contaminated samples or positive results of the samples are further confirmated with technique of liquid chromatography system connected with the double mass spectrometry LC-MS/MS. The implementation of validation for the determination of aflatoxins in tree nuts, ELISA technique made the basis for continuous monitoring of aflatoxin in this type of food. Laboratory has the capabilities to the rapid and reliable way to determine whether a contaminated food which is essential when it comes to imports of such food in Croatia