Isolated fractions from Tragopogon dubius showed enhanced antiproliferative activity against colon cancer cell line (HCT-116), GCHRMS and OHRLCMS profiling

Tragopogon dubius is traditionally consumed as a vegetable and employed in folk medicine for the treatment of inflammatory diseases, skin conditions, and cutaneous disorders. Little is known about the pharmacological investigations associated with its vegetative part and roots, with many of its in-depth biological attributes and active phytochemicals remaining unexplored. This study investigates the phytochemical composition, antioxidant activity, and anticancer properties of crude hexane extract (TrDH) from leaves and stems of Tragopogon dubius and isolated fractions. Partition coefficient crystallization and Preparative thin-layer chromatography were utilized to separate the crude extract into isolated Cr-MTH, Cr-CHL, TrDH-1, TrDH-2, and TrDH-3 fractions from T. dubius to evaluate comparative antioxidant and anticancer properties. Gas chromatography High Resolution Mass Spectrometry (GCHRMS) and Orbitrap High Resolution Liquid Chromatography Mass Spectroscopy OHRLCMS analysis revealed a variety of bioactive compounds, including terpenes, flavonoids, diterpenes, fatty acids, alkaloids, and phenolic acids. Antioxidant activity was quantitatively assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and superoxide radical scavenging assays, with TrDH-3 demonstrating strong activity, exhibiting IC50 values ranging as 71.64 μg/mL, 68.79 μg/mL, and 106.37 μg/mL, respectively. The anticancer potential was evaluated against the Colon cancer cell line (HCT-116), revealing that TrDH-3 significantly inhibited cell proliferation, with a GI50 (50% Growth Inhibitory concentration) value as low as 38.87 μg/mL. Fluorescence microscopy indicated that TrDH-3 induces apoptosis, as evidenced by alterations in nuclear morphology and loss of mitochondrial membrane potential. These findings demonstrate that T. dubius hexane extract and its fractions, particularly the TrDH-3 fraction, possess significant antioxidant and anticancer properties. This study highlights the potential of T. dubius as a valuable source of natural bioactive compounds with promising health benefits.