constants from reacting the lightest and the heaviest hydrogen analogues with 1H2 were then used to calculate the k0.11/k4.1 kinetic isotope effect, in which there is a 36.4 fold difference in isotopic masses. For this reaction, isotopic substitution happens to produce an inverse kinetic isotope effect, and the authors report a kinetic isotope effect as low as 1.74 x 10−4, which is the smallest kinetic isotope effect ever reported. The kinetic isotope effect leads to a specific distribution of deuterium isotopes in natural products, depending on the route they were synthesized in nature. By NMR spectroscopy, it is therefore easy to detect whether the alcohol in wine was fermented from glucose, or from illicitly added saccharose. Another reaction mechanisms that was elucidated using the kinetic isotope effect is the halogenation of toluene: In this particular "intramolecular KIE" study, a benzylic hydrogen undergoes radical substitution by bromine using N-bromosuccinimide as the brominating agent. It was found that PhCH3 brominates 4.86x faster than PhCD3. A large KIE of 5.56 is associated with the reaction of ketones with bromine and sodium hydroxide. In this reaction the rate-limiting step is formation of the enolate by deprotonation of the ketone. In this study the KIE is calculated from the reaction rate constants for regular 2,4-dimethyl-3-pentanone and its deuterated isomer by optical density measurements. In asymmetric catalysis, there are rare instances in which a kinetic isotope effect manifests as a significant difference in the enantioselectivity observed for a deuterated substrate compared to a non-deuterated one. One example was reported by Toste and coworkers, in which a deuterated substrate produced an enantioselectivity of 83% ee, compared to 93% ee for the undeuterated substrate. The effect was taken to corroborate additional inter- and intramolecular competition KIE data that suggested cleavage of the C-H/D bond in the enantiodetermining step. See also Crossover experiment (chemistry) Equilibrium constant#Effect of isotopic substitution Isotope effect on lipid peroxidation Kinetic Isotope Effects of RuBisCO (ribulose-1,5-bisphosphate carboxylase oxygenase) Magnetic isotope effect Reaction mechanism Transient kinetic isotope fractionation Urey-Bigeleisen-Mayer equation References Further reading Organic chemistry Physical organic chemistry Chemical kinetics Reaction mechanisms