Abstract: A two-dimensional preparative liquid chromatographic method was developed for purification of constituents from water extract of Armillaria luteo-virens using reversed phase preparative liquid chromatography(RPLC)coupled with hydrophilic interaction liquid chromatography(HILIC). Seven compounds were isolated from fruit bodies of Armillaria luteo-virens, and their structures were identified on the basis of physicochemical and spectroscopic analyses. These compounds were identified as pyroglutamic acid( 1 ), uridine( 2 ), 2′-deoxyuridine( 3 ), uracil( 4 ), guanosine( 5 ), inosine( 6 ), and adenosine( 7 ). All the compounds were purified and identified from fruit bodies of Armillaria luteo-virens for the first time.