β-carotene is a carotenoid pigment that converts to vitamin A in human body and plays crucial role in visual functions, reproductive performance and immune system. Jute leaves contain β-carotene but amounts in different varieties of jute leaves are not known. The study was undertaken to estimate the β-carotene content in fresh jute leaves of the cultivated jute species Corchorus olitorius L. and Corchorus capsularis L. by spectrophotometry and thin layer chromatography. β- carotene content varies in leaves of different varieties of two species. Leaves of some C. olitorius variety showed higher amount of β-carotene than the leaves of C. capsularis variety. Some varieties of C. olitorius showed similarity with some varieties of C. capsularis. Variety Robi-1 of C .olitorius showed the highest content of β-carotene 39.93mg/100g. O-72, O- 4, O-3820, O-9897 and OM-1 of olitorius variety showed higher β-carotene than all the varieties of C. capsularis. JRO- 524 and MG-1 of C. olitorius variety is comparable to CVL-1, D-154, BJC-2197 and deshi pat shak-1 variety of C. capsularis regarding β-carotene content. Lowest β-carotene was estimated 12.55mg/100g in O-795 of olitorius variety and 12.32mg/100g in BJC-83 and 11.34mg/100g in BJC-2142 of capsularis variety. Thin layer chromatography system silica gel on TLC Al foils as stationary phase and hexane as mobile phase performs good separation and identification of β- carotene in jute leaves.