Abstract: To explore the applicability and rationality of zebrafish model for the evaluation of in vivo anti-osteoporosis activity of micro-amount of components from traditional Chinese medicine, zebrafish larvae osteoporosis model induced by 25 μmol/L prednisolone was used to test the anti-osteoporosis activity of asperosaponins V and VI. Zebrafish bones were stained with alizarin red, and quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. The results indicated that 25 μmol/L prednisolone could make zebrafish bone lose significantly, and that compared with the model group, asperosaponins V and VI and etidronate disodium could prevent such loss induced by prednisolone. The results suggested that zebrafish model could be used in evaluating anti-steoporosis activity of micro-amount of asperosaponins V and VI, which has advantages of being in vivo, micro plate, simple and highly efficient.