Mozhgan Soltani - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Kazem Parivar - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Javad Baharara - Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
Mohammad Amin Kerachian - Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
Javad Asili - Department of Pharmacognosy, School of Pharmacy, Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

Background: Holothuroids (sea cucumbers) are members of the phylum echinodermata, which produce saponins. Saponins exhibit a wide spectrum of pharmacological and biological activities. In this study, we isolated the crude saponins from the body wall of the dominant Iranian species of sea cucumber, Holothuria leucospilota (H. leucospilota). The purpose of this study was to confirm the presence of saponins in the Persian Gulf H. leucospilota and study the hemolytic and cytotoxic activities of these compounds. Methods: The body wall of sea cucumber was dried and powdered and the crude saponins were isolated using various solvents. The crude saponins were further purified by column chromatography using HP-۲۰ resin. The foam test, Thin Layer Chromatography (TLC), hemolytic assay, and Fourier Transform Infrared Spectroscopy (FTIR) confirmed the presence of saponins. Cytotoxicity was analyzed using a ۳-(۴, ۵-dimethylthiazol-۲-yl)-۲, ۵-diphenyltetrazolium bromide (MTT) assay on A۵۴۹ cells, a human lung cancer cell line. Results: The foam test, hemolytic assay, and TLC supported the presence of saponin compounds in the ۸۰% ethanol fraction of H. leucospilota. The infrared (IR) spectrum of the extract showed hydroxyl (-OH), alkyl (C-H), ether (C-O) and ester (–C=O) absorption characteristic of teriterpenoid saponins. The C-O-C absorption indicated glycoside linkages to the sapogenins. The crude saponin extracted from sea cucumber was cytotoxic to A۵۴۹ cells. Conclusion: The ۸۰% ethanol fraction of saponin isolated from H. leucospilota exhibited hemolytic activity and offers promise as an anti-cancer candidate.

Cytotoxicity assay, Hemolytic assay, Holothuria leucospilota, Saponin, Sea cucumber