Objective: To optimize preparation process of standard extract from Coptidis Rhizoma and establish quality control method of it. Method: L9(34) orthogonal experiment was used to optimize the extraction process of CoptidisRhizoma. The contents of berberine hydrochloride

palmatine hydrochloride and coptisine hydrochloride were detected simultaneously by HPLC. Result: The optimal condition for extraction were as follows: most coarse powder of Coptidis Rhizoma was reflux extracted with 10 times 60% ethanol for 1 h

for three times. The extraction yield was 25.47%. The contents of berberine hydrochloride

palmatine hydrochloride and coptisine hydrochloride were 23.02%

5.63% and 5.72%

respectively. The calibration curve of berberine hydrochloride was linear in the range from 8.5 to 271.2 mg ·L-1 (r>0.999 9) and the average recovery was 100.14% (RSD 1.92%). The calibration curve of palmatine hydrochloride was linear in the range from 4.2 to 133.2 mg ·L-1 (r>0.999 9) and the average recovery was 98.52% (RSD 1.65%). The calibration curve of coptisine hydrochloride was linear in the range from 3.1 to 97.2 mg ·L-1 (r>0.999 9) and the average recovery was 99.80% (RSD 1.45%). Conclusion: Reflux extraction with ethanol was effective method for preparation of standard extract form Coptidis Rhizoma. HPLC methods established can be used for quality control of the standard extract.