Daun srikaya (Annona squamosa L., Annonaceae) digunakan dalam pengobatan tradisional sebagai penurun kadar gula darah. Simplisia yang berasal dari dua lokasi tumbuh dibandingkan kandungan kimianya. Penapisan fitokimia kedua simplisia, karakteristik simplisia, dan pola kromatogram ekstrak menunjukkan hasil yang mirip. Simplisia diekstraksi secara refluks menggunakan pelarut dengan kepolaran meningkat, yaitu n-heksana, etil asetat, dan metanol. Ekstrak n-heksana salah satu simplisia disaponifikasi, difraksinasi secara kromatografi cair vakum, dilanjutkan dengan kromatografi kolom. Fraksi dipisahkan menggunakan kromatografi lapis tipis preparatif dan isolat dikarakterisasi menggunakan spektrofotometri ultraviolet-sinar tampak dan spektrofotometri inframerah serta penampak bercak Liebermann-Burchard. Isolat merupakan triterpenoid dengan gugus fungsi O"“H, CH, dan C=C serta tidak memiliki ikatan rangkap terkonjugasi.Kata Kunci: Annona squamosa L., daun Srikaya, triterpenoid Sugar apple (Annona squamosa L., Annonaceae) leaf has been traditionally used to treat high blood glucose level. Crude drugs obtained from plants growing at two different locations were studied to evaluate whether there were differences in their phytochemical constituents. Phytochemical screening of those two crude drugs, the quality, and extract chromatographic pattern showed similar results. Crude drugs were extracted by reflux using n-hexane, ethyl acetate, and methanol. N-hexane extract from one of the crude drugs was saponified, followed by fractionation using vacuum liquid chromatography and column chromatography. The fractions were separated using preparative thin layer chromatography and one isolate was characterized using Liebermann-Burchard reagent, ultraviolet-visible and infrared spectrophotometry. The isolate was a triterpenoid with O-H, C-H, and C=C groups and did not have any conjugated double bond.Keywords: Annona squamosa L., sugar apple leaf, triterpenoi