Inhibitory effects of 130 medicinal plants on hyaluronidase activity were analyzed. The medicinal plants are clinically used as herbal medicines for korean traditional prescriptions. Six out of the 130 herbal medicines exhibited more than 50% of inhibition on hyaluronidase activity by their total methanol extracts with 5mg/ml as a final concentration. The active total methanol extracts were prepared from cortex of Acantbopanax gracilistylus, lignum of Caesalpinia sappan, radix of Glycyrrhiza uralensis, radicis cortex of Morus alba, herba of Prunella vulgaris, and radix of Sanguisorba officinalis. These active total methanol extracts were sequentially fractionated with dichloromethane, ethyl acetate, n-butanol, and then water. Among the solvent-fractionated extracts, the butanol fractions of Acantbopanax gracilistylus and Glycyrrhiza uralensis with 1 mg/ml as the final concentration exhibited more than 50% of inhibition on hyaluronidase activity, and the other fractions with the same concentration did less than 20% of inhibition.