The leaves extracts of natural and commercial used tobacco sample tested on UV, IR spectrum and phytochemical screening. The colour of the natural tobacco extract was dark green, it’s shape was mucilaginous and has pH 8.35, while the colour of the commercial tobacco extract was dark brown, it’s shape was mucilaginous and has pH = 7.6. Furthermore, percentage yield of the raw extract of tobacco leaves plant and the raw extract of commercial tobacco were 12.723 and 7.341%, respectively. While percentage yield of alkaloids for tobacco leaves and alkaloids for commercial, tobacco was 6.32 and 4.13% correspondingly. The results of the qualitative detection tests revealed the presence of alkaloids in natural tobacco and commercial tobacco extracts. The UV spectrum of nicotine from Tubacam nicotina corresponds to the wavelength of the maximum absorption peak (λ Max of the compound) with the radiation spectrum of the standard compound also. Although the UV spectrum of nicotine form, commercial tobacco corresponds to the wavelength of the maximum absorption peak (λ Max of the compound) with the radiation spectrum of the standard compound. Furthermore, the absorption of nicotine in the infrared region, revealed to be identical to the IR absorption of the Nicotine standard, at 2917 cm where the vibration frequencies of the most critically active groups in the compound.