~ 283 ~ Journal of Pharmacognosy and Phytochemistry 2022; 11(4): 283-294 E-ISSN: 2278-4136 P-ISSN: 2349-8234 www.phytojournal.com JPP 2022; 11(4): 283-294 Received: 23-05-2022 Accepted: 29-06-2022 Lali Lingfa Department of Genetics, Osmania University, Telangana, India Baloji Gugulothu Department of Genetics, Osmania University, Telangana, India Sandhya Jagtap Department of Genetics, Osmania University, Telangana, India Srinivas Ankanagari Department of Genetics, Osmania University, Telangana, India Corresponding Author: Srinivas Ankanagari Department of Genetics, Osmania University, Telangana, India HPTLC fingerprinting reveals leaf and roots phytochemical variability in developmental stages of Withania somnifera Lali Lingfa, Baloji Gugulothu, Sandhya Jagtap and Srinivas Ankanagari DOI: https://doi.org/10.22271/phyto.2022.v11.i4d.14473 Abstract Background: Withania somnifera has been the foundation of numerous contemporary medicines today for treatment of various diseases. However, phytochemical variability profile is lacking in the root and leaf organs. Aim: This study was aimed to elucidate the phytochemical profile in seedling, vegetative and reproductive stages of leaf and roots in W. somnifera using High performance thin layer chromatography (HPTLC) fingerprinting. Methods: The HPTLC with dual wavelength UV (254/366 nm) was used for the analysis of phytochemical variation in leaf and root methanolic extracts of W. somnifera. Results: HPTLC chromatograms in 366 nm, 254 nm, and normal day light, seedling, vegetative and reproductive stages leaf and root exhibited bands variability. At 450 nm leaf HPTLC fingerprints in seedling stage showed five peaks with Rf values in the range of 0.42 to 0.90, vegetative stage with nine peaks of Rf values in range of 0.10 to 0.89, and reproductive stage with six peaks of Rf values in range of 0.30 to 0.92. In root showed six peaks with Rf values in range of 0.00 to 0.82 in seedling stage, ten peaks with Rf values in range of 0.00 to 0.96 in vegetative stage, seven peaks with Rf values in range of 0.01 to 0.91 in reproductive stage. In roots, higher peak areas were found at the reproductive stage (3.33%) compared to vegetative stage (1.65%). Conclusion: The comparative HPTLC fingerprints of leaves and roots in W. somnifera presented phytochemical variability in leaf and root that varied with developmental stages. Keywords: HPTLC fingerprinting, Withania somnifera, leaf, root, phytochemicals, herbal medicine 1. Introduction Herbal medicines have been actively utilized in the health care system of Indian history since immemorial time. But its growth as an industry is still not explored to its full potential (Ekor, 2014; Petrovska, 2012; Pandey et al., 2013) [1, 2, 3] . However, rates of its production have been increased noticeably in the past few years (Jang et al., 2017) [4] . Hence, there is a need to transmute the herbal medicine into a vibrant scientifically authenticated and evidence based industrial products, based on deep understanding of traditional medicine and available research (Mohiuddin, 2019) [5] . In fact, it is important to use methods for rapid and precise identification and estimation of specific stage of medicinal plants and the essential phytoconstituents, as the qualitative and quantitative objectives to assess the genuineness and crucial quality (Balekundri and Mannur, 2020; Kira nmai, 2016) [6, 7] . The estimation and evaluation of herbal medicinal plants, its phytochemicals through an accurate method would offer to find the true essential phytochemicals in herbs used for therapeutic purposes and provide a precise foundation of herbal medicine effectiveness (Pan et al., 2013; Jasemi et al., 2020) [8, 9] . The identification and estimation can be achieved through various analytical techniques including HPTLC (Attimarad et al., 2011; Patel et al., 2011) [10, 11] . Such analytical techniques will ascertain the presence of essential phytochemical constituents and quantify their concentrations (Sonia et al., 2017) [12] . HPTLC offers many advantages over other various analytical methods of chromatographic techniques. It can present the results in an image, autonomous, reliable, easier to handle, many samples for parallel analysis, low in experimental expenses, rapid presentation of the results, and detection sensitivity of phytoconstituents in picogram (pg) and nanogram (ng) concentrations (Kharat et al., 2017; Nikolin et al., 2004; Bairy, 2015) [13, 14, 15] .