Research On The Physiological Active Substances From Styela Clava

Nowadays, research on the physiological active substances from the marine tunicates is in a high speed progress. But it hardly can be seen on papers that some tunicate extracts which have high effective anticancer potential are obtained from Styela clava'?, tunica and body or other kinds of substances. And the research work carried on Styela Cla\>a in homeland is rarely reported. Our country especially coastal areas in Shandong province abounds tunicates, But comparing with the overseas, the resources greatly disturb the cultivation and sailing, not mention they are utilized. Therefore we should pay great attention to them. In this paper, we made a basic research on the physiological active substance from Styela clava in following aspects stated below. I . Research on liposoluble constituentFirst, Styela clava's tunica and its body were separated carefully so that interactions among some substances are prevented. In this paper, we focus mainly on the chemical compound from the tunica of Styela clava. According to the frequently used methods of extraction and isolation of natural products from organic body, the freeze-dried ascidian tunica was exhaustively extracted with 100% chloroform then the combined extracts were filtered and the solvent was separately concentrated under vacuum to yield a dark-brown gum. This material was loaded onto a silica gel column and the column was eluted with chloroform several times until the fraction of interest was collected. Finally we obtained some light yellow green ointment liquid F I .Second, we used high performance liquid chromatography (HPLC), thin-layer chromatography(TLC), gel chromatography (GC), and protein and nucleic acid analytic apparatus to evaluate F I ' s purification. All data indicated that F I was a pure substance. Third, using modern analytic techniques such as nucleic magnetic resonance(NMR), infrared spectroscopy (IR), ultraviolet visible absorption spectra (UV) and mass spectrometry(MS), we analyzed the basic chemical structure of F I and nominated it phthalic acid di (3-methyl) nanyl ester (PDNE). We estimated primarily it was a novel natural compound.Fourth, we investigated the PDNE's anticancer activity by culturing cancer cells such asB37 mammary cancer cell and BEL7402 liver cancer cell through MTT method. From the data of the experiment we found that PDNE was able to inhibit the activity of BEL7402 liver cancer cell significantly under the concentration 40 u g/ml and this effect was concentration-dependent. But the effect of this compound on B37 mammary cancer cell with the same concentration was very mild. We believe that research on physiological substances in this field in our country will lead to a rapid progress through further investigation about this compound. II. Research on water-soluble constituentWe also did some research on the water-soluble physiological active constituent from the tunica of Styela clava. The method we used to extract and purify the water-soluble one was somewhat different from that we used in liposoluble PDNE. As for the water-soluble constituent, analysis about purification and structure on it was still not carried out, and only some basic research on its anticancer activity was finished. Data from the experiment indicate that the green extract HQLF could inhibit the activity of B37 mammary cancer cell.We believe that through further research, the work in this field in our country will make a rapid progress and a branded-new prospect will come out.