Methylene diphenyl diisocyanate (MDI) and toluene diisocyanate (TDI) are high production volume chemicals used for the manufacture of polyurethanes. For both substances, the most relevant adverse health effects after overexposure in the workplace are isocyanate-induced asthma, lung function decrement and, to a much lesser extent, skin effects. Over the last two decades many articles have addressed the reactivity of MDI and TDI in biological media and the associated biochemistry, which increased the understanding of their biochemical and physiological behavior. In this review, these new insights with respect to similarities and differences concerning the adsorption, distribution, metabolism, and excretion (ADME) of these two diisocyanates and the implications on their toxicities are summarized. Both TDI and MDI show very similar behavior in reactivity to biological macromolecules, distribution, metabolism, and excretion. Evidence suggests that the isocyanate (NCO) group is scavenged at the portal-of-entry and is not systemically available in unbound reactive form. This explains the lack of other than portal-of-entry toxicity observed in repeated-dose inhalation tests.