Objective To investigate the water soluble chemical constituents in the root of Valeriana officinalis Linn. var. latiofolia Miq. and to identify their structures. Methods The water soluble chemical constituents were isolated and purified by macroporous resin Diaion HP-20, silica gel and Sephadex LH-20; the chemical structures of the components were determined by MS, 1D NMR and 2D NMR spectral analysis. Results and conclusion Thirteen constituents were isolated and identified from the root of Valeriana officinalis Linn. var. latiofolia, including Ganxinoside A (1), (-)-angelicoidenol-2-O-β-D-glucopyranoside (2), (-)-angelicoidenol-2-O-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (3), 6-hydroxy-7-(hydroxymethyl)-4-methylenehexahydrocyclopenta[c]pyran-1 (3H)-one (4), kanokoside A (5), prinsepiol-4-O-β-D-glucopyranoside (6), (+)-pinoresinol-4-O-β-D-glucopyranoside (7), sinenoside Ⅰ (8), lanicepsides A (9), (+)-1-hydroxypinoresinol-1-O-β-D-glucoside (10), (+)-pinoresinol-4, 4'-O-bisglucopyranoside (11), (+)-cycloolivil 6-O-β-D-glucopyranoside (12), and coniferin (13). All the 13 compounds are isolated from the root of Valeriana officinalis Linn. var. latiofolia Miq. for the first time.