AIM:To identify the optimal extraction of anti-asthma active protein from fresh Earthworm(Eisenia foetida).METHODS:Extraction time,extraction solvent and pH were selected as influencing factors.90 quinea pigs were randomly divided into 9 groups(control,montelukast,low temp,room temp,acidic buffer extract,alkaline buffer extract,aqueous extract,neutral buffer extract,powder earthworm group).Anti-asthma effciency was compared among groups.extraction time,saturation of ammonia sulfate buffer volume were selected as factors of orthogonal design.RESULTS:The optimal extraction for anti-asthma protein was at 4 ?C,adding 2 fold volume of pH 6.8 buffer phosphate,homogenating,standing and extracting for 3 hours,taking the supernataut and centifuging,salting out with 80% ammonia sulfate,suspecting with 20 mmol/L,pH6.8 buffer phosphate.So Earthworm protein was gained from the clear supernatant.CONCLUSION:The extraction technique may keep its original activity.