title: Hemoglobin adducts and urine metabolites of 4,4'-methylenedianiline after 4,4'-methylenediphenyl diisocyanate exposure of rats.
abstract: 4,4'-Methylenediphenyl diisocyanate (MDI) is a very important component in the production of polyurethane. In a long-term experiment, designed to determine the carcinogenic and toxic effects of MDI, rats were exposed chronically for 3 and 12 months, to 0.0 (control), 0.26, 0.70 and 2.06 mg MDI/m3 as aerosols. Hemoglobin adducts and urine metabolites of MDI were determined at the different doses in order to develop methods to biomonitor workers exposed to MDI and to assess a risk resulting from such exposure. Hemoglobin adducts and urine metabolites of 4,4'-methylenedianiline (MDA) were found in all rats, including controls. MDA and N-acetyl-MDA (AcMDA) were quantified by GC-MS after derivatization with heptafluorobutyric anhydride. The dose-response relationships for hemoglobin adducts and urine metabolites were non-linear over this dose range. In urine, free AcMDA and MDA were found after base extraction. The amount of MDA present in urine and to a lesser extent the AcMDA found in urine correlate well with the corresponding amount determined as hemoglobin adducts for all dose groups. In order to release MDA from possible conjugates of MDA and AcMDA, urine was treated under strong acidic conditions. Following this procedure higher MDA levels were found than the sum of MDA and AcMDA from mild base hydrolysis. Similar results were obtained with the rats exposed for 3 and 12 months, indicating that a steady state had been reached by 3 months. In order to perform further investigations of the bronchoalveolar lavage fluid one group of animals was given a 1 week recovery period before sacrifice. Hemoglobin adducts from these animals showed a decrease of approximately 40% for all dose groups. According to the lifetime of rat erythrocytes the levels of hemoglobin adducts should have decreased by only 22%. This suggests that the erythrocytes with modified hemoglobin have a shorter lifespan. In order to exclude the possibility that hemoglobin adducts may have resulted from ingestion of hydrolyzed MDI via licking of the fur, a single dose experiment with rats exposed through the nose only or with the whole body was carried out. The only difference observed between these two exposure regimes was that the hemoglobin adduct levels of AcMDA after nose only exposure were significantly higher than after total body exposure. The presence of AcMDA in urine and as a hemoglobin adduct indicates that MDA was bioavailable after MDI exposure. The presence of MDA may contribute significantly to the carciongenic potential of MDI, since MDA has been shown to be carcinogenic in animals.
authors.affiliation: Institut für Pharmakologie und Toxikologie, Universität Würzburg, Germany.
authors.forename: O
authors.initials: O
authors.lastname: Sepai
authors.forename: D
authors.initials: D
authors.lastname: Schütze
authors.forename: U
authors.initials: U
authors.lastname: Heinrich
authors.forename: H G
authors.initials: HG
authors.lastname: Hoymann
authors.forename: D
authors.initials: D
authors.lastname: Henschler
authors.forename: G
authors.initials: G
authors.lastname: Sabbioni
mesh_terms: D000083:Acetanilides / Q000097:blood / Q000652:urine; D000280:Administration, Inhalation; D000814:Aniline Compounds / Q000097:blood* / Q000652:urine*; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D005260:Female; D006454:Hemoglobins / Q000378:metabolism; D017953:Isocyanates / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D001559:Benzhydryl Compounds
chemical_list: D000083:Acetanilides; D000814:Aniline Compounds; D006454:Hemoglobins; D017953:Isocyanates; C050075:N-acetyl-4,4'-diaminodiphenylmethane; C005969:4,4'-diphenylmethane diisocyanate; D001559:Benzhydryl Compounds; C009505:4,4'-diaminodiphenylmethane