Differential metabolites identified by untargeted metabolomics in midgut samples of Helicoverpa armigera larvae injected with dsβGRP4 and dsGFP.

Midguts were harvested from dsGFP and dsβGRP4 injected larvae at 6th 24 h, with five biological replicates for each group. Midgut metabolites were extracted, vortexed, and subjected to protein precipitation.

Following centrifugation, supernatants were dried under nitrogen gas, re solubilized, and re centrifuged for purification. Metabolite profiling was implemented via ultra performance liquid chromatography coupled to a Q Exactive mass spectrometer (Thermo Fisher Scientific, MA, USA), using an ACQUITY HSS T3 column (Waters, Milford, USA). Metabolite annotation and downstream data processing were performed against the Human Metabolome Database.