Browsing by Subject "MPLC"

Rhazya stricta Decne. is a small evergreen shrub belonging to the Apocynaceae family. The plant grows in South Asia and the Middle East, and in these areas it is used in traditional medicine. All parts of the plant are used in different preparations for a variety of purposes such as infections, bowel diseases, itching and diabetes. R. stricta synthesizes about a hundred different alkaloids, of which only a fraction has been studied closer. Some of the analyzed alkaloids have showed some interesting pharmacological properties such as antibacterial and cytotoxic properties. Because it is often both economically and ecologically unsustainable to cultivate or to collect large amounts of medicinal plants from nature, cell cultures have been developed from plants. The properties and synthesized substances of the cell cultures can be analysed and modified in laboratories. In the experimental part of this work, a system was developed for alkaloid extraction, fractionation and isolation from dried cell material from cultured R. stricta hairy root-cells. The goal was to develop a functioning system that eventually enables identification of the alkaloids synthesized by the cultured cells under given conditions. Alkaloids were extracted from 26 g of dried and ground cell mass. The fractionation of the alkaloids was performed with medium pressure liquid chromatography (MPLC) and the fractions were analyzed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The alkaloids were purified by horizontal TLC and preparative HPLC. Ion-pair chromatography was used for analyzing the extract, fractions and purified alkaloids. Five components from two fractions were eventually isolated. One of the components was tentatively identified as vincanine, but further analyzes have to be performed to identify all components reliably. In total, hairy root-cells seem to synthesize approximately 20 alkaloids with variable polarity.