ConspectusThis Account presents a new discipline, single sample molecular chronology (SSMC), which studies the relative age of an individual compound occurring in several temporal pools of a single sample in complex media. Geochemists have analytically observed for a long time that several pools of the same compound, e.g., a hydrocarbon or a pesticide, can be isolated from the same sample, e.g., a sediment or a soil, to yield a free compound pool obtained by solvent extraction and then a bound compound pool after treatment of the solid residue and further extraction. Yet the study on the significance of these pools has been limited due to the inherent lack of criteria to clearly distinguish the same compound present in various pools, and, as a consequence, the existence of these pools has been criticized as resulting from a default of extraction during analytical fractionation. Our breakthrough was to distinguish isotopically several temporal pools of a plant-derived C31 n-alkane in a soil sample containing naturally 13C-labeled carbon and then to set up a method, 13C-relative dating, to calculate the relative age of these temporal pools. We observed wide differences in the relative age of the C31 n-alkane in temporal pools of a single soil sample, ranging from -6.7 years for a soil humin-bound homologue to +25.1 years for the free homologue in the coarser soil particle-size fraction. Individual compounds can thus be used as molecular clocks to determine the relative age of temporal pools from the same sample. Moreover, our findings represented the first unambiguous proof that bound compounds are cycling slower and are somehow protected in a complex organo-mineral matrix, key information for the mechanism of carbon sequestration. SSMC could be developed in all disciplines of physical, biological, and environmental sciences manipulating complex media, to study the history of individual compounds. This chronochemistry should provide new information about the origin and transformation of individual compounds in biogeochemical systems. For example, historical information on drugs or pollutants encapsulated in temporal pools of a living organism would bring about critical new knowledge about the mechanisms of disease development. Investigations require isotope tracing using any isotope in natural or artificial abundance. Methods to separate temporal pools are suggested.