Abstract: Tri-(4-methylbenzoate) cellulose chiral stationary phase (MCTB) was prepared. The separation of enantiomers of 2-(4-hydroxyphenoxy) propionate had been investigated by reversed-phase high-performance liquid chromatography on the chiral stationary phase. The results showed that the separation of enantiomers of methyl, ethyl and butyl esters of 2-(4-hydroxyphenoxy) propionic acid could be achieved on the chiral stationary phase (MCTB) using methanol and water(volume ratio is 75:25) as mobile phase. Their separation factors were 1.38, 1.49, 0.98, respectively. At the same time,It was found that the size of the ester group obviously affected the separation efficiency of their enantiomers on the MCTB chiral stationary phase. The separation of the ethyl ester was most satisfactory.