Clostridium botulinum type C strain Stockholm (C-St) produced two types of progenitor toxins (M and L). The purified L toxin consisted of neurotoxin (NT), nontoxic-nonhemagglutinin (NTNHA) and hemagglutinin (HA) components which could be separated into four subcomponents, HA-33, HA-17, HA-22-23 and HA-55. On the other hand, purified M toxin lacked HA components. We isolated all components from the progenitor L toxin by following chromatographic methods. The L toxin was separated into an NT and NTNHA/HAs complex via Mono Q anion exchange column chromatography. Using gel filtration with a TSK-gel HW-55S column in the presence of 6M guanidine hydrochloride, the NTNHA/HAs complex derived from the L toxin could be separated into three fractions, NTNHA single component, HA-55/33 mixture and HA-22-23/17 mixture. Then, the HA-55/33 mixture was separated into each component using Mono S cation exchange column chromatography in the presence of 8M urea. The HA-22-23/17 mixture was also separated from each other with Superose 12 gel filtration column chromatography in the presence of 8M urea. The separated NT, NTNHA and HA subcomponents were homogenous on SDS-PAGE, and were identified to be C-St progenitor toxin gene products based on their N-terminal amino acid sequence analysis. This study presents for the first time an attempt to isolate components from the progenitor toxin. Although reassociation of the isolated components was also attempted via mixing of NTNHA, HA-55, HA-33, HA-22-23 and HA-17, HA activity of the mixture was lower than that of parent NTNHA/HAs complex.