Flavonoids profile for the medicinally important plant Euphorbia neriifolia (Sehund) was established by using high performance thin layer chromatography (HPTLC). For this direct and sequential soxhlet extraction were performed and then fractions were concentrated and subjected to HPTLC. The derivatization of developed plates was done by exposing to ammonia and iodine vapours. The plates were photo-documented at UV 366 nm and daylight using photo-documentation chamber. In the current investigation the benzene extract of E. neriifolia leaves illustrated the presence of 5 different types of flavonoids with Rf values ranging from 0.06 to 0.96. The chloroform extract of E. neriifolia demonstrated the presence of 3 to 9 different types of flavonoids with Rf values ranging from 0.03 to 0.96. The ethyl-acetate extract of E. neriifolia demonstrated the presence of 6 to 12 different types of flavonoids with Rf values ranging from 0.2 to 0.97 and ethanol, aqueous and hydro-ethanol extracts of E. neriifolia demonstrated the presence of 5-12 different types of flavonoids with different Rf values ranging from 0.032 to 0.95. Maximum number of flavonoids were observed in ethylacetate and aqueous extract (12) followed by minimum in benzene and chloroform (3-7) extracts of leaves of E. neriifolia. In the present study we observed various flavonoid profile of the E. neriifolia using HPTLC.