CAT assay question

[Harold Drabkin]

Subject: CAT assay question


E>Following the TLC of a CAT reaction, four (or a subset of four) spots might
E>be visualzed.  These correspond to unacetylated chloramphemicol, two
E>different species of mono-acetylated chloramphenicol, and di-acetylated
E>chloramphenicol.  The presence of di-acetylated chloramphenicol probably
E>indicates that the reaction has preceeded too far, and is not useful for
E>quantitation.

E>My question is:  When calculating the "percent conversion" for a particular
E>reaction, do you take into account both mono-acetylated species, or just
E>the top most form (the more abundant form)?  Is the issue actually just a
E>matter of consistency in your number crunching from assay to assay?

I always include the two monospecies together.  One of them (the faint
one) is due to (as I remember) a non-enzymatic rearrangement.  Thus, it
is valid to do so.  If the reacton gives diacetyl, it's off linear range
anyway.

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