New Isocyanate-Specific Albumin Adducts of 4,4′-Methylenediphenyl Diisocyanate (MDI) in Rats

Abstract

4,4'-Methylenediphenyl diisocyanate (MDI) is the most important of the isocyanates used as intermediates in the chemical industry. Among the main types of damage after exposure to low levels of MDI are lung sensitization and asthma. Albumin adducts of MDI might be involved in the etiology of sensitization reactions. It is, therefore, necessary to have sensitive and specific methods for monitoring the isocyanate exposure of workers. To date, urinary metabolites or protein adducts have been used as biomarkers in workers exposed to MDI. However, with these methods it is not possible to determine whether the biomarkers result from exposure to MDI or to the parent aromatic amine 4,4'-methylenedianiline (MDA). This work presents a procedure for the determination of isocyanate-specific albumin adducts. In a long-term experiment, designed to determine the carcinogenic and toxic effects of MDI, rats were exposed chronically for 3 months, to 0.0 (control), 0.26, 0.70, and 2.06 mg MDI/m(3) as aerosols. Albumin was isolated from plasma, digested with Pronase E, and analyzed by LC-MS/MS. MDI formed adducts with lysine: N(6)-[({4-[4-aminobenzyl]phenyl}amino)carbonyl]lysine (MDI-Lys) and N(6)-[({4-[4-(acetylamino)benzyl]phenyl}amino)carbonyl] lysine (AcMDI-Lys). For the quantitation of the adducts in vivo, isotope dilution mass spectrometry was used to measure the adducts in 2 mg of albumin. The adducts found in vivo (MDI-Lys and AcMDI-Lys) and the corresponding isotope labeled compounds (MDI-[(13)C(6)(15)N(2)]Lys and Ac[(2)H(4)]MDI-Lys) were synthesized and used for quantitation. The MDI-Lys levels increased from 0-24.8 pmol/mg albumin, and the AcMDI-Lys levels increased from 0-1.85 pmol/mg albumin. The mean ratio of MDI-Lys/AcMDI-Lys for each dose level was greater than >20. The albumin adducts correlate with other biomarkers measured in the same rats in the past: urinary metabolites and hemoglobin adducts released after mild base hydrolysis. This method will enable one to measure isocyanate-specific albumin adducts in workers. This new biomonitoring procedure will allow for the assessment of suspected exposure sources and may contribute to the identification of individuals who are particularly vulnerable for developing bronchial asthma and other respiratory diseases after exposure to isocyanates. In addition, it will help to improve the production of antigens for the analysis of antibodies in exposed workers.