Methods of separating the stereoisomers of secondary di- and tri-alcoholic compounds comprise reacting a racemic mixture of the secondary alcohols with an acyl anhydride or an ester in the presence of an enzyme which stereoselectively acylates the (R) site of the alcohol to form the (R) site acyl ester of the alcohol and separating the (S,S) or (S,S,S) secondary alcohol, the (R,S) or (R,S,S) monoacyl ester of the alcohol, the (R,R) or (R,R,S) diacyl ester and/or the (R,R,R) triacyl ester of the alcohol from one another and from the reaction medium. Additional methods of separating the stereoisomers of a secondary di- or tri-alcohol comprise reacting a stereochemical mixture of acyl esters of the di- or tri-alcohol with an enzyme which selectively deacylates the (R) or (S) site of the secondary alcohol to produce (R) or (S) site deacylated isomers of the secondary alcohol and separating the free alcohol, the monoacyl ester, the diacyl ester, and the triacyl ester of the alcohol from one another and the reaction medium.