Readily available l-menthyl phthalate has been shown to be an effective derivatizing agent for determination of the enantiomeric purity of alkyl- and aryl-substituted 1,1,1-trifluoromethyl-2-alkanols using HPLC and GC. It has been shown that a previously described protocol for one-step enzymatic kinetic resolution results in the formation of the desired 1,1,1-trifluoromethyl-2-alkanols with 96–98% ee. Enrichment of the (R)-isomer of trifluoromethyl alkanols by repetition of the enzymatic hydrolysis procedure was found to increase the ee up to 99.9%. Furthermore, excessive conversion of the corresponding esters during enzymatic hydrolysis allowed enantiomerically pure (S)-1,1,1-trifluoromethyl-2-alkanols to be obtained.