eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2025-03-25 22 1 367 374 10.13005/bbra/3368 54636 article Leela Karunakaran 1 Anita Rosemarie Joseph Singh 1 Department of Biotechnology, Women’s Christian College, Affiliated to the University of Madras, Chennai, Tamilnadu, India Plant parts contain diverse distribution of secondary compounds which plays a predominant role in promoting health and well-being along with vital roles in environmental and agricultural applications. In the present study, crude ethanol extracts prepared from Mangifera indica L., Piper betle L. and Lawsonia inermis L., leaves were subjected towards purification by column chromatography using hexane, chloroform, ethyl acetate and methanol in 4:3:2:1 ratio as column mobile phase. The obtained purified fractions were further resolved in thin layer chromatographic plates with hexane and ethyl acetate in 7.2:2.8 ratio as mobile phase. Best fraction from each of the three samples showing better separation was characterized by means of gas chromatographic – mass spectrometric analysis for determining the existence of valuable as well as pharmacologically active components. Totally 15 bioactive compounds were identified in each of these purified fractions wherein, 3 compounds were observed to be similar in the fractions of all three samples and includes Meso-2,3-Bis-T-Butoxysuccinonitrile; Methyl 3-Methyl-5-Oxy-2-Phenoxyhexanedithioate and 2,3,4,4-Tretrapropyl-1-(Trimethylsilyl)-1-(Trimethylsilyloxy)-1,3-Diaza-2,4-Diborabutane. Further investigation on these compounds could pave way towards development of drugs of therapeutic and industrial importance. https://www.biotech-asia.org/vol22no1/purification-and-identification-of-secondary-compounds-in-mangifera-indica-l-piper-betle-l-and-lawsonia-inermis-l-leaves-by-column-chromatography-and-gc-ms-analysis/ GC-MS - Gas Chromatography -  Mass Spectrometry; R_f – Retention Factor; TLC – Thin Layer Chromatography; UV – Ultra Violet