The ability of three anti-juvenile hormone agents (AJHAs) to prevent the juvenile hormone dependent phase of larval-pupal transformation has been investigated. Neck-ligated post-feeding last instar larvae of Spodoptera mauritia were treated topically with different doses of a juvenile hormone analog (JHA) or the AJHAs, tetra-hydro-4-fluoromethyl-4-hydroxy-2H-pyran-2-one (FMev), ethyl-4-[2-(tert-butyl carbonyloxy)butoxy] benzoate (ETB) and ethyl-[E]-3-methyl-2-dodecanoate (EMD). Untreated ligated larvae or those treated with acetone survived for 12 ± 1 days without showing any sign of pupal cuticle secretion. On the other hand treatments of ligated larvae with different doses of JHA induced pupation. Among the three AJHAs tested treatments of ETB promoted pupation. Treatments of ligated larvae with lower doses of EMD induced the formation of larval-pupal intermediates whereas those treated with higher dose moulted into either pupae or larval-pupal intermediates. FMev-treated larvae exhibited a complete inhibition of moulting and metamorphosis. In order to study whether AJHAs would prevent the JHA induced pupation, neck-ligated larvae were treated simultaneously with different doses of AJHAs and 1 μg JHA. Co-application of JHA with different doses of EMD induced pupation in majority of the ligated larvae and thus appears to a certain extent to counteract the effects of treatments of same doses of EMD alone. Co-application of JHA with different doses of ETB elicited pupation in all the surviving ligated larvae. To a large extent co-application of JHA with different doses of FMev failed to induce pupation. The significance of the findings is discussed in the context of the mode of action of these AJHAs.