The present invention relates to a method of inactivating enveloped viruses in a viral preparation predominantly containing non-enveloped viruses by the action of a solvent at a temperature of between -5 C. and +50 C. and at a pH of between about 5 and 9. Its subject is also a method of preparing a viral preparation comprising such a method of inactivation. The invention also relates to a viral preparation obtained according to the method of the invention. Finally, it relates to a host cell and a composition comprising such a viral preparation as well as their uses for therapeutic or prophylactic purposes.