The fact that glutathione (GSH) plays many roles in biological protective mechanisms and critical physiological functions has been recognized for decades. Conjugates, disulfides, and other glutathione-derived products also have been studied as biomarkers of the chemical natures or specific identities of key metabolites of toxic agents and such studies have been crucial in the delineation of the nature of the interactions of proximal toxicants with target biomolecules. Despite the extensive evidence implicating the depletion and/or oxidation of glutathione in a wide variety of human and experimental toxicities, critical examination of such studies frequently reveals that injury is not simply related to glutathione status. GSH is compartmentalized at several levels and this compartmentation appears to exert considerable influence on the relationships between glutathione depletion or oxidation and the onset of injury. Although compartmentation is usually viewed from the perspective of different intracellular pools, the significance of extracellular glutathione in functionally important pools is gaining recognition. As the factors affecting the interactions of intracellular pools with extracellular pools are delineated, studies in humans can be designed and interpreted with greater precision and utility.