A novel process for 6-aminnopenicillanic acid (6-APA) production from penicillin G fermentation broth was developed, which integrated some new technologies such as three-liquid-phase extraction and solvent sublation. A new three-liquid-phase extraction system containing ethylene oxide-propylene oxide (EOPO) random copolymer was used to separate and purify penicillin G from its filtrated broth and trace organic extractant in the aqueous phase of back-extraction was removed by solvent sublation. Experimental results showed that above 90% of penicillin G could be distributed into the upper organic phase at pH below 3.0, and most of impurities and some phenylacetic acids (PAA) were partitioned into the copolymer-rich middle phase. About 70% organic solvent could be stripped by one-step solvent sublation from penicillin G aqueous solution. The purity of 6-APA produced by the above-simplified process could be above 98% and other quality parameters reached the level of the conventional process. (c) 2005 Elsevier Ltd. All rights reserved.