Background: Roots of Solanum nigrum Linn. (Solanaceae), commonly known as black night shade (Kakamachi), is traditionally used in the treatment of worms and abdominal pain. Until date no scientific evaluation has been reported on its roots. Aim: The present study deals with the microscopical, histochemical, physicochemical, fluorescence analysis, preliminary phytochemical and chromatographic study of roots of S. nigrum L. Materials and Methods: Thin sections of fresh root were used for the microscopical and histochemical evaluation. Root powder was used for physicochemical and fluorescence analysis. For preliminary phytochemical study water, methanol and ether soluble extractives were used. Chromatographic study was carried out on methanol soluble extractive. Results: Diagnostic features of the roots are presence of prismatic crystals, border pitted vessels, and tannin content. Physico‑chemical analysis of root powder shows ash value 3.99% w/w, 13.04% w/w and 11.85% w/w of water and alcohol soluble extractive values respectively. Phytochemical investigations of root shows presence of alkaloids, tannins and saponins along with other phytoconstituents in different extracts. In chromatographic study, alcoholic extract of root shows 10, 7 and 8 number of spots at 256 nm, 366 nm wavelengths and after spray of vanillin sulphuric acid, respectively. Conclusion: The information generated in this study will be helpful for the proper identification and authentication of roots of this herb. Key words: Kakamachi, pharmacognosy, phytochemistry, Solanun nigrum Linn