Kinetic Isotope Effects of RuBisCO carbon, oxygen, and/or hydrogen, though currently only a significant isotope effect is seen for carbon isotope substitution. Isotopes are atoms that have the same number of protons but varying numbers of neutrons. "Lighter" isotopes (like the stable Carbon-12 isotope) have a smaller overall mass, and "heavier" isotopes (like the stable Carbon-13 isotope or radioactive Carbon-14 isotope) have a larger overall mass. Stable isotope geochemistry is concerned with how varying chemical and physical processes preferentially enrich or deplete stable isotopes. Enzymes like RuBisCO cause isotopic fractionation because molecules containing lighter isotopes have higher zero-point energies (ZPE), the lowest possible quantum energy
Reference materials for stable isotope analysis is almost always the isotopologue with no isotopic substitutions. This isCO for carbon dioxide and CH for methane. Standard isotopic reference materials are still required in clumped isotope analysis for measuring the bulk δ values of a sample, which are used to calculate the expected stochastic distribution and subsequently to infer clumped isotope temperatures. However, the clumped isotope composition of most samples are altered in the mass spectrometer during ionization, meaning that post-measurement data correction requires having measured materials of known clumped isotope composition. At a given temperature equilibrium thermodynamics predicts the distribution of isotopes among possible isotopologues, and these