An improved method for the extraction of mutagens from human urine and cooked meat using blue rayon.
A reproducible method has been developed and validated that allows the detection of mutagenic material in human urine following the intake of a meal containing pan-fried beef patties. The mutagens are extracted from the urine with blue rayon and eluted with methanol/ammonia (100:1). Using 14C-2-amino-3-methylimidazo[4,5-f]quinoline (14C-IQ) as a tracer, the extraction efficiency of heterocyclic amines was consistently found to exceed 90%. It was observed that the critical factor in attaining high extraction efficiencies was the concentration of blue rayon, which depended on the nature of the matrix. Constituents of human urine were demonstrated to compete with IQ for the binding sites on the blue rayon. For the determination of the mutagenic activity in the extracted urine, the O-acetylase over-expressing Salmonella typhimurium strain YG1024 was utilised, and the Ames test procedure was optimised with respect to S9 concentration and incorporation of a pre-incubation step. Using the developed procedure, a marked rise in mutagenic activity, determined both in the absence and presence of an activation system, was evident in human urine following the intake of a beef patty meal. Normal mutagenicity levels were restored within 24 h. Finally, a method is described, once again employing blue rayon, for the extraction of heterocyclic amines from cooked beef patties with a 92% efficiency, as exemplified by IQ.
Yoxall
Victoria
V
Molecular Toxicology Group, School of Biomedical and Molecular Sciences, University of Surrey, Guildford, Surrey GU2 7XH, UK.
Wilson
Judith
J
Ioannides
Costas
C
D000818:Animals; D001667:Binding, Competitive; D002250:Carbon Radioisotopes / Q000378:metabolism; D002417:Cattle; D002482:Cellulose / Q000378:metabolism; D003296:Cooking; D006571:Heterocyclic Compounds / Q000032:analysis; D006801:Humans; D008460:Meat / Q000032:analysis*; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000032:analysis*; D011804:Quinolines / Q000378:metabolism; D012486:Salmonella typhimurium; D014556:Urine / Q000737:chemistry*
D002250:Carbon Radioisotopes; D006571:Heterocyclic Compounds; D009153:Mutagens; D011804:Quinolines; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; D002482:Cellulose; C012024:rayon, purified