Hayvan yemlerinde aflatoksin tayini
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Dosyalar
Tarih
2015
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Aflatoksinler, önemli tarımsal ürünlerde sentezlenebilen (kabuklular, tahıllar vb.) küf metabolitleridir. İnsan ve hayvan sağlığı için önemli sağlık etkilerine neden olabilirler. Bu çalışmada, hayvan yemlerinde aflatoksinlerin kantitatif tayini için, asetonun (% 55/45, % 70/30, % 85/15), metanolün (% 60/40,% 70/30,% 80/20), asetonitrilin (% 55/45, % 70/30, % 85/15) farklı karışımları kullanıldı. Metodun validasyonu için sertifikalı referans madde (hayvan yemi) ve spike yapılmış örnekler (hayvan yeminde toplam aflatoksinin farklı konsantrasyonları olarak; 1.3-2.6-5.2-10.4-15.6 µg/kg ) kullanılmıştır. Belirlenen HPLC şartlarında (?ex = 360 nm, ?em = 430 nm, akış hızı: 1.0 mL/dk, kolon sıcaklığı: 25 0C, enjeksiyon hacmi: 100 µL, C18 kolon, mobil faz: su/metanol/asetonitril (56/26/18) 120 mg KBr ve 350 µL 4M HNO3 içeren) örnekler analiz edildi. Aflatoksin B1, B2, G1 ve G2’ nin geri alma oranları, %70/30 metanol/su karışımında kantitatif olarak bulunmuştur. Çalışma sonunda, seksen adet hayvan yemi örneğinin beş adedinde 10-20 µg/kg ve bir adet örnekte 122.51 µg/kg aflatoksin B1 tespit edilmiştir. Yirmi dokuz adet örnekte aflatoksin B1 tespit edilememiştir. Örneklerin aflatoksin B1 ortalaması 4.78 µg/kg’ dır. Sadece 1 adet örnek (122.51 µg/kg) yasal limiti aşmıştır.
Abstract
Aflatoxins are fungal metabolite that can be synthesized in important agricultural products (nuts, cereals, etc.) For human and animal health, aflatoxins cause important health effects. In this study, different mixtures of acetone (% 55/45, % 70/30, % 85/15), methanol (% 60/40, % 70/30, % 80/20), acetonitrile (% 55/45, % 70/30, % 85/15), used for the quantitative determination of aflatoxins in animal feed. Certified reference material ( animal feed) and spiked samples (different consentrations of total aflatoxin at animal feed: 1.3-2.6-5.2-10.4-15.6 µg/kg ) are used for method validation. At optimed conditions for HPLC (?ex = 360 nm, ?em = 430 nm, flow rate: 1.0 mL/min, column temperature: 25 0C, injection volume: 100 µL, C18 column, mobil phase: water/methanol/acetonitrile (56/26/18) with 120 mg KBr and 350 µL 4M HNO3 ) samples were analysed. Recoveries of aflatoxin B1, B2, G1 and G2 were stable when samples were extracted with %70/30 methanol/water mixture. In this study, aflatoxin B1 were detected with 10- 20 µg/kg at five sample and 122.51 20 µg/kg at one samples. At tewenty nine of eighty sample aflatoxin B1 not detected. Mean aflatoxin B1 concentration of samples is 4.78 µg/kg. Only one sample (122.51 µg/kg) exceeded the legal limit of 20 µg/kg aflatoxin B1.
Abstract
Aflatoxins are fungal metabolite that can be synthesized in important agricultural products (nuts, cereals, etc.) For human and animal health, aflatoxins cause important health effects. In this study, different mixtures of acetone (% 55/45, % 70/30, % 85/15), methanol (% 60/40, % 70/30, % 80/20), acetonitrile (% 55/45, % 70/30, % 85/15), used for the quantitative determination of aflatoxins in animal feed. Certified reference material ( animal feed) and spiked samples (different consentrations of total aflatoxin at animal feed: 1.3-2.6-5.2-10.4-15.6 µg/kg ) are used for method validation. At optimed conditions for HPLC (?ex = 360 nm, ?em = 430 nm, flow rate: 1.0 mL/min, column temperature: 25 0C, injection volume: 100 µL, C18 column, mobil phase: water/methanol/acetonitrile (56/26/18) with 120 mg KBr and 350 µL 4M HNO3 ) samples were analysed. Recoveries of aflatoxin B1, B2, G1 and G2 were stable when samples were extracted with %70/30 methanol/water mixture. In this study, aflatoxin B1 were detected with 10- 20 µg/kg at five sample and 122.51 20 µg/kg at one samples. At tewenty nine of eighty sample aflatoxin B1 not detected. Mean aflatoxin B1 concentration of samples is 4.78 µg/kg. Only one sample (122.51 µg/kg) exceeded the legal limit of 20 µg/kg aflatoxin B1.
Açıklama
Doktora Tezi
Anahtar Kelimeler
Aflatoksin, HPLC, İmmunoaffiniti, Ekstraksiyon, Aflatoxin, Immunoaffinity, Extraction
