Acetoin is a common food flavor additive. This volatile compound widely exists in nature. Some microorganisms, higher plants, insects, and higher animals have the ability to synthesize acetoin using different enzymes and pathways under certain circumstances. As a very active molecule, acetoin acts as a precursor of dozens of compounds. Therefore, acetoin and its derivatives are frequently detected in the component analysis of a variety of foods using gas chromatography-mass spectrometry. Because of the increasing importance of these compounds, this paper reviews the origins and natural existence of these substances, physiological roles, the biological synthesis pathways, nonenzymatic spontaneous reactions, and the common determination methods in foods. This work is the first review on dietary natural acetoin.