Abstract: CBZ-phenylalanine was reacted with 6-OH of β -cyclodextrin,then the derivative was bonded to silica to produce a new chiral stationary phase (CSP) for high performance liquid chromatography(HPLC). The derivative of β -cyclodextrin and the CSP were characterized by Molisch color reaction, IR and XPS.Chromatographic performance of the bonded silica CSP was evaluated using benzene and naphthalene as testing materials. The m -, o -, p-isomers of nitrophenol and aminophenol were separated well on the column of the CSP. Enantiomers of clenbuterol hydrochloride and 5,6-O-isopropyl-3-thio-hexanoic acid-1,4-lactone were investigated and tolerably separated on the column.