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More specifically, the present invention relates to a process for preparing L-carnitine or acetyl L-carnitine hydrochloride, comprising sequentially synthesizing racemic 4-chloro-3-hydroxybutyronitrile of Formula 2 and racemic 4-chloro-3-hydroxy butyric acid alkyl ester of Formula 3 under specific reaction conditions, using racemic epichlorohydrin of Formula 1 as a starting material, preparing (R)-4-chloro-3-hydroxy butyric acid alkyl ester of Formula 4 from stereoselective hydrolysis of the racemic 4-chloro- 3 -hydroxy butyric acid alkyl ester in an aqueous solution using an enzyme, and preparing L-carnitine of Formula 6 or acetyl L-carnitine hydrochloride of Formula 7 from the (R)-4-chloro-3-hydroxy Carnitine exists as two distinct isomers, i.e