Isolation, Purification and Identification of Cocin and Kaempferol, Powerful Antioxidants in Crocus sativus L. Flowers.

Crocus sativus L. is an autumn-flowering geophyte extensively grown and cultivated in countries such as Italy, Spain, Greece, Turkey, Iran and India. Commercially used saffron is dried stigma which has been used for some of its physiologic properties for centuries. Recently, attention has been paid to the identification of new sources of safe natural antioxidants from Crocus sativus flowers that are normally wasted. The antioxidant activities are mainly attributed to their carotenoid and flavonoid compounds, like Crocin and Kaempferol. These bioactive compounds have important biological activities; cancer and prevention and treatment of heart, vascular disease. Crocin is a diester formed from the disaccharide gentiobiose and the dicarboxylic acid crocetin and is known to inhibit proliferation and nucleic acid synthesis. It induces apoptosis in the human tongue squamous cell carcinoma cell line Tca8113 and hepatocarcinoma. Kaempferol is a natural flavonoid. Numerous reports have shown that Kaempferol and/or Kaempferol glycosides induce cell death in a variety of cancer cells from different tissues. Many of these reports have shown that it is effective in reducing vascular endothelial growth factor expression in ovarian and colon cancer cells. Extraction of these antioxidants was carried out using methanol and ethanol in different concentrations for each compound. Isolation and purification, in comparison with authentic standards, was achieved by Flash Column Chromatography and Thin Layer Chromatography (TLC). Crocin and Kaempferol were chemically characterised by infrared (IR), mass spectroscopy (MS) and nuclear magnetic resonance (1H & 13C NMR). The in-vitro cytotoxicity towards cancer cell lines will also be investigated. The presence of these important groups of medicinal natural products in saffron’s flowers that were considered waste, now paves the way of new biological effects.