C-3Ha are also relatively small, frequently necessitating the use of special techniques. Substitution by heavier isotopes can also give rise to a isotope effects, but they are frequently too small to be of great value in enzyme mechanistic work.

One of the main applications of secondary isotope effects is to distinguish Sn1 and Sn2 reactions. The a-kinetic isotope effect for Sn1 reactions is typically significantly larger than that observed in SN2-type reactions. Such behavior reflects changes in the geometry of the carbon atom undergoing substitution:

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