Komparacija ELISA i TLC/HPLC metoda za određivanje ZEA i OTA u žitaricama i krmi

Scientific paper - Original scientific paper

• Title (english): COMPARISON BETWEEN ELISA AND TLC/HPLC METHODS FOR DETERMINATION OF ZEARALENON AND OCHRATOXIN A IN FOOD AND FEED
• Author: Maja Šegvić Klarić
• Author: Stjepan Pepeljnjak
• Author: Zdenka Cvetnić
• Author: Ivan Kosalec
• Author's institution: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of microbiology)
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract (croatian)

Fusarium i Penicillium vrste plijesni javljaju se kao najučestaliji kontaminanti žitarica u području umjerene klime. Kao posljedica se mikotoksini zearalenon (ZEA) i okratoksin A (OTA) često nalaze u takvim supstratima u većim ili manjim koncentracijama ovisno o načinu pospremanja usjeva, skladištenju i mikroklimatskim uvjetima. Cilj rada je bio odrediti pojavu ZEA i OTA tijekom 2007. godine u nasumično uzimanim uzorcima žitarica i krmiva (N=37) u individualnim domaćinstvima na području endemske nefropatije (EN) u Hrvatskoj. Za određivanje ZEA i OTA korišten je kompetativni direktni ELISA test (Veratox_) prema preporukama proizvođača Neogen Europe Ltd. te modificirana metoda visokoučinkovite tekućinske kromatografije (HPLC) prema Frisvad i Thrane (1987) i tankoslojna kromatografija (TLC). Usporedbom metoda određene su granice detekcije i kvantifikacije te korelacije sadržaja ispitivanih mikotoksina. ZEA je dokazan u 91,9% uzoraka u rasponu od 12,5 µg/kg do 1182 µg/kg (srednja koncentracija 318,3 µg/kg), dok je OTA nađen u 16,2% uzoraka s rasponom od 2,5 µg/kg do 31,7 µg/kg (srednja koncentracija 9,8 µg/kg). Kokontaminacija je dokazana u 13,5 % uzoraka. Utvrđene vrijednosti ZEA i OTA u hrani i krmivu su ispod najvećih dopuštenih količina prema Pravilniku (Narodne novine br. 118. iz 2007.) te odgovaraju i preporukama Europske komisije (EC/576/2006) i EC/1881/2006). Usporedbom metoda, utvrđeno je da najnižu granicu detekcije ima ELISA, a slijedi ju HPLC. Izmjerene koncentracije mikotoksina statistički značajno koreliraju između ELISA i HPLC metode. Prednost ELISA metode je u jednostavnosti izvedbe (pogotovo za veliki broj uzoraka), osjetljivosti, ekonomskoj isplativosti te ekološkoj prihvatljivosti. Prednost HPLC metode je u analizi više mikotoksina u jednom uzorku.

Abstract (english)

Fusarium and Penicillium are the most common mould species that contaminate cereals in the mild climatic zone. Therefore, zearalenone (ZEA) and ochratoxin A (OTA) are frequently detected in these substrates in different concentrations depending on storage and microclimatic conditions. The purpose of this study was to determine the frequency and concentrations of ZEA and OTA in cereal and feed samples randomly collected in individual households of endemic nephropathy region (EN) in Croatia. Direct competitive ELISA test kit was used for ZEA and OTA determination according to manufacturer instructions. Results of ELISA method as well as limits of detection (LOD) and quantification were compared with high performance liquid chromatography (HPLC) and thin layer chromatography (TLC) methods. Correlation between results was also analysed. ZEA was detected in 91.9% of samples ranging between 12.5 and 1168µg/kg (mean 318.3 µg/kg), while OTA was recovered from 16,2% of samples in concentrations ranging from 2.5 µg/kg to 31.7 µg/kg
(mean 9.8 µg/kg). ZEA and OTA simultaneously occurred in 13.5 % of samples. Detected values of mycotoxins were under the permissible levels according to Croatian law (Narodne novine No. 118/2007 and European Commission (EC/576/2006 and EC/1881/2006). The lowest LOD was obtained by ELISA followed by HPLC. Mycotoxin concentrations detected by ELISA significantly correlate with results obtained by HPLC. ELISA provide an attractive and promising method for OTA and ZEA detection in food due to high specificity, sensitivity, simplicity (analysis of high number of samples) and it is ecologically more acceptable than chromatographic methods. Advantage of HPLC method is multiple mycotoxin detection and quantification in one sample.

• Keywords (croatian): ELISA i TLC/HPLC metode; zearalenon; okratoksin
• Keywords (english): ELISA i TLC/HPLC methods; zearalenon; ochratoxin
• Language: croatian
• Publication type: Scientific paper - Original scientific paper
• Publication status: Published
• Peer review: Peer review - domestic
• Publication version: Published version
• Journal title: Krmiva : Časopis o hranidbi životinja, proizvodnji i tehnologiji krme
• Numbering: vol. 50, no. 5, pp. 235-244
• p-ISSN: 0023-4850
• e-ISSN: 1848-901X
• URN:NBN: urn:nbn:hr:163:458223
• Publication: 2008-12-22
• Document URL: https://hrcak.srce.hr/32670
• Type of resource: Text
• Access conditions: Open access
• Created on: 2018-02-27 12:16:11