Abstract: OBJECTIVE Toxicity model of zebrafish was used to evaluate safety of 26 kinds of common orthopedic herbal medicine. METHODS Zebrafish embryo at 1 day post fertilization (dpf) were exposed with various concentration of water decoction; the death number of the embryos or larvals was counted from 1dpf to 6 dpf; embryonic micro-morphology of zebrafish (3dpf) was observed and pictures were taken that compared with 0.4% DMSO; LC₅₀ value at 6dpf was calculated by SPSS16.0. RESULTS The results indicated that 13 kinds of water decoction of herbal medicines (Dipsaci Radix， Anemarrhena Rhizoma， Olibanum and Myrrha， Psoraleae Fructus， Angelicae Pubescentis Radix， Achyranthis Bidentatae Radix， Polygoni multiflori Radix， Cnidii Fructus， Aucklandiae Radix， Eupolyphaga Steleophaga， Epimedii Folium and Acanthopanacis Cortex) can cause obvious organ toxicity to juvenile zebrafish， such as yolk sac swelling， deformation， black， pericardial edema and bleeding， and their LC₅₀ values were low (16.9~689.7 μg/mL)， while the other 13 herbal medicines are relatively safe (Pyrolae Herba， Homalomena Rhizoma， Sappan Lignum， Morindae Officinalis Radix， Testudinis Carapax Et Plastrum， Drynariae Rhizoma， Salviae miltiorrhizae Radix Et Rhizoma， Cibotii Rhizoma， Taxilli Herba， spatholobi CauLis， Rehmanniae Radix Praeparata， Ligustri Lucidi Fructus and Eucommiae Cortex)， no fish organ toxicity was observed， and their concentration caused zebrafish death was usually at high levels (above 2 000 μg/mL). CONCLUSION Zebrafish model was successfully used for screening toxicity of 26 common orthopedic herbal medicine， which may provide useful information for reasonable application of them. Zebrafish model has advantages of simplicity， high efficiency and real-time observation， which made large scale toxicity screening of herbal medicine in vivo style possible.