Five kilograms of tobacco hornworm eggs (48 to 64 hours old) afforded 26.5 milligrams of a new crystalline insect molting hormone identified as 26-hydroxyecdysone. The three known insect ecdysones-alpha-ecdysone, 20-hydroxyecdysone, and 20,26-dihydroxyecdysone-were also present but in much smaller quantities. The new hormone is the predominant molting hormone in the hornworm during this stage of embryonic development. These results takent in context with the current knowledge of the chemistry and biochemistry of the molting hormones during postembryonic development in the hornworm indicate quantitative and qualitative differences in the biosynthetic-mnetabolic pathways as well as in the ecdysones in different developmental stages of this insect.