Delphinium is a plant rich in alkaloids belonging to the Diterpenoid Alkaloids that contain medicinal properties. The aim and importance of this study was to separate and purify the compound using classical and modern chromatography. In addition to use modern spectroscopic studies to determine the structure of the new alkaloid produced from this type of plant located in Syria. The new C20-Diterpenoid Alkaloid (Atisine skeleton) Delphitisine was isolated from the aerial parts of Delphinium peregrinum L. var. eriocarpum Boiss. (collected from Amm Harten village between Homs and Tartous cities at the end of July, and identified by different chromatographic methods). It is colourless resin 31mgr. The structure was established mainly by IR, EI-Ms, 1D NMR spectroscopy: (1H NMR, 13C NMR, DEPT-135) and 2D NMR spectroscopy: (DQF-COSY, HMQC). © 2020. All Rights Reserved.