Cr-IV 1 PLA₂ (D49) was purified from Calloselasma rhodostoma venom after two chromatographic steps, molecular exclusion on Superdex G-75 and reverse phase HPLC on m-Bondapack C-18 (0.8 X 30 cm) (Waters Corp.). Cr-IV1 it has evidenced a local myotoxic activity “In vivo”. A new PLA₂ Cr-IV1 induced a conspicuous local myotoxic effect when injected by the in route, but no increase in CK plasma levels occur after iv injection. Time-course analysis showed a maximum increase in CK 2 hour after injection, and returned to normal by 24 h. Experiments were carried out in triplicate. The results show that the Cr-IV1 PLA₂ produce local myotoxicity non systemic in smooth muscle of mouse in different ways. The Cr-IV1 in the 10 mg/100 µl dose is unable to produce local myotoxicity. These results show that the enzymatic activity could be related to the “in vivo” local myotoxic effect in smooth muscle of mouse. Our results probably showed the binding to many cell types in tissues and blood. Therefore Cr-IV1 is not distributed to muscle tissue after intravenous injection, and non increments in serum CK activity in these conditions are reduced when compared with serum CK activity after intramuscular administration (Gutiérrez and Ownby, 2003; Kini 2003). For the edematogenic activity it was prepared a stock solution of 1mg/mL of which were prepared doses from Cr-IV1 of 0.5; 2.5; 5.0 and 10 mg/ml. This solutions were inoculated via subplantar to groups of four mice from 18 to 20 g weight. The fraction Cr-IV1 show edematogenic activity, dependent dose and time-course, decreased after 24 hours. Our result show a local effect, characteristic of the viperides venom.