eng Oriental Scientific Publishing Company Biomedical and Pharmacology Journal 0974-6242 2023-03-21 16 1 275 288 10.13005/bpj/2609 47530 article Partha Saha 1 Dipshikha Sharma 2 Suvakanta Dash 3 Kumar Saurav Dey 4 Samir Kumar Sil 1 Cancer Biology and Cell Physiology Lab., Department of Human Physiology, Tripura University, Suryamaninagar-799022, Tripura, India. Assam Science and Technology University, Guwahati-781013, Assam, India. Regional Institute of Pharmaceutical Science and Technology, Agartala-799005, Tripura, India. Guwahati Biotech Park, Guwahati-781039, Assam, India. Parkia javanica (Lamk.) Merr. is an ethnomedicinal leguminous plant species from northeastern India with a long history of medicinal use among various tribes of this region to treat cholera, dysentery, stomach aches, diarrhea and food poisoning, having antibacterial, wound-healing, anticancer and anti-inflammatory properties. Therefore, in this current study, the methanolic bark extract was carried out and fractionated by using flash chromatography, examined the cytotoxicity of the respective fractions on colon cancer cell lines, and evaluated the major phytochemical compounds present in the fractions using Gas Chromatography-Mass Spectrometry (GC-MS) chemical profiling. Chemical profiling of the fractions by GC-MS revealed in fraction-1 and -2, 2,4-Di-tert-butylphenol was the major compound (50.740% in fraction-1, 21.277% in fraction-2, and 7.859% in fraction-3) having reported anticancer activity. The gradation of the presence of this compound in the fractions was corroborated by the gradation of anti-colon cancer activity of the respective fractions on both the colon carcinoma cell lines. However, the presence of D-Allose in a substantial amount (20.870%) in only fraction-3 could not increase the anticancer activity of fraction-3 over the other two fractions. An in vitro cytotoxic assay guided evaluation of three flash chromatographic fractions (fraction-1, -2, and -3) of methanolic extract of Parkia javanica bark showed significant anticancer properties on two human colon carcinoma cell lines (HCT116 and SW480). The order of efficacy of the fractions was fraction-1> fraction-2 > fraction-3. In a time and dose-dependent experiment, fraction-1, being the most active one, showed an IC₅₀ value of 16.25 µgml^-1 (24 hrs), 9.94 µgml^-1 (48 hrs), and 9.38 µgml^-1 (72 hrs) on HCT116 and 35 µgml^-1 (24 hrs), 20.14 µgml^-1 (48 hrs), and 19.71 µgml^-1 (72 hrs) on the SW480 cell line. Parkia javanica bark extract is bestowed with the potential of anti-colon cancer property and upon chemical profiling of different chromatographic fractions of the extract, 2,4-Di-tert-butylphenol has been identified as the primary anticancer component of the extract. https://biomedpharmajournal.org/vol16no1/identi%ef%ac%81cation-of-24-di-tert-butylphenol-24-dtbp-as-the-major-contributor-of-anti-colon-cancer-activity-of-active-chromatographic-fraction-of-parkia-javanica-lamk-merr-bark-extract/ Anti-Colon cancer  Bark extract  2 4-Di-tert-butylphenol  Flash chromatography  GC-MS  Parkia javanica