Discovery of the F2-isoprostanes, a group of prostaglandin F2-like compounds biosynthesized from arachidonic acid nonenzymatically, has uncovered a new and novel facet of free radical biology. Some of these compounds are bioactive and thus may mediate adverse effects associated with oxidant stress. F2-Isoprostanes have also been shown to be reliable biomarkers of lipid peroxidation. Factors influencing their formation and metabolism have been studied to some extent, although much remains to be determined. The purpose of this review is to summarize our current knowledge of conditions that modulate endogenous generation of these compounds. Isoprostanes have a wide daily variation in secretion in humans. Although normal levels can be defined, these compounds are found in increased concentrations in various pathophysiological states, including ischemia-reperfusion injury, atherosclerosis, and diabetes, and in experimental conditions of oxidative stress and inflammation. Alterations in isoprostane biosynthesis, secretion, and excretion in normal physiology and in pathophysiological states are due to the various types of endogenous and exogenous regulatory mechanisms that control the availability of precursors required for isoprostane synthesis, such as dietary and tissue arachidonic acid content, oxygen concentration, and the generation of various free radical species. Selected aspects of issues related to isoprostane formation and metabolism in vivo will be examined herein.