DL-2-methyl-1-butanol was resoluted by porcine pancreatic lipase(PPL)with tributyrin served as substrate and organic phase.Tributyrin,2-methylbutyl butyrate and dibutyrin in the sample from the reaction process were separated by TLC.Their respective spots were obtained by using developing solution of hexane:ether:ethyl-acetate:methanol(10:2:1:1 in vol.)and placing spreaded gel panel in iodine steam.The spots were analyzed by a TLC analyzing software designed by Visual C++programming language,with satisfactory accuracy.It shows that monitoring the reaction by TLC is realizable.