Abstract. It is expected that information on the source, reaction
pathway, and reaction kinetics of an organic compound can be obtained from
its position-specific isotope compositions or intramolecular isotope
distribution (Intra-ID). To retrieve the information, we could use its
predicted equilibrium Intra-ID as a reference for understanding the observed
Intra-IDs. Historically, observed, apparently close-to-equilibrium carbon
Intra-ID has prompted an open debate on the nature of biosystems and
specifically the pervasiveness of reversible biochemical reactions. Much of
the debate remains unresolved, and the discussion has not clearly
distinguished between two states of equilibrium: (1) the equilibrium among the
corresponding bond-breaking and bond-forming positions in reactant and product and
(2) the equilibrium among all carbon positions within a compound. For an
organic molecule with multiple carbon positions, equilibrium carbon Intra-ID
can be attained only when a specific reaction is in equilibrium and the
sources of each position are also in equilibrium with each other. An
observed Intra-ID provides limited information on if the sources and pathways
are both unconstrained. Here, we elaborate on this insight using examples of
the observed Intra-IDs of hydroxyl-bearing minerals, N2O, and acetic
acid. Research effort aiming to calibrate position-specific equilibrium
and kinetic isotope fractionation factors for defined processes will help to
interpret observed Intra-IDs of a compound accurately and fully.