Sophora tonkinensis is a herb used in traditional Chinese medicine. Sofalcone is an oral gastrointestinal medication and a synthetic derivative of sophoradin, an isoprenyl chalcone found in S. tonkinensis. A new matrine-type alkaloid, (-)-14 beta-hydroxyoxymatrine (1), was isolated from the roots and rhizomes of Sophora tonkinensis Gapnep. (Leguminosae), together with five known matrine-type alkaloids, (-)-14 beta-hydroxymatrine (2), (+)-oxymatrine (3), (+)-matrine (4), (+)-sophoranol (5), and (-)-5 alpha-hydroxysophocarpine ( 6), as well as with two known cytisine-type alkaloids, (-)-cytisine (7) and (-)- N-methylcytisine (8). Their structures were elucidated by spectroscopic methods. Compounds 1, 5 and 7 showed potent anti-HBV activity with an inhibitory potency against HBsAg secretion of 22.6 %, 31.1 % and 33.2 %, and against HBeAg secretion of 30.4 %, 26.3 % and 27.8 %, respectively.