ISOLATION AND CHARACTERIZATION OF THE STRONGEST CYTOTOXICITY AGENT FROM MARRUBIUM ASTRACANICUM

Abstract

The pharmacological appraisement of constituents from plants is an established technique for the identification of lead compounds which can leads to the development of novel and secure medicinal agents. We selected Marrubium astracanicum (Lamiaceae) to explore for its bioactive constituents. The objective of this study was to isolate potential cytotoxicity agent from M. astracanicum crude methanol extracts from the flowering shoots, using a bioassay-guided strategy. For this purpose, we used brine shrimp lethality test (BST) that is attractive method and very useful tool for the isolation of bioactive compounds from plant extracts because it is very simple, reliable and low toxin amounts are sufficient to perform the test in the microwell scale and also is confirmed by the National Cancer Institute (NCI). The crude extract has the most effect with LC50 performed by column chromatography with n-hexane, ethyl acetate and methanol solvents, respectively. Ethyl acetate fraction with LC50 most active component. By using thin layer chromatography (TLC), HPLC-UV analytical method, gradient elution of column chromatography with n-hexane and ethyl acetate (and subjected to sequential fractionation) and BST, ultimately the most effective compound was isolated and purified. This compound displays LC50 values of 4 f equal to 0.21 in chloroform:methanol (150:1, v/v). The separation efficiency of compound B from total extracts calculated 0.99 percent. The compound B with the scientific name ' Bis(1-pentyl) disulfide' completely identified by using spectroscopy techniques of 13C-NMR, 1H-NMR, HCOSY, IR and UV. The present study reveals two results; it is clear that the isolation and purification of plant extracts provides bioactive components that not only eliminate ineffective or even harmful substances, but also increases their effectiveness. On the other hand, we are facing with plant that shows great cytotoxicity. These two factors are critical from the perspective of biological activity and medicinal properties, especially in the field of anticancer [1,2]