| Aims.Valeriana jatamansi Jones is the root and rhizome of the plant Valeriana jatamansi Jones of Valeriana Linn of Valerianaceae.The earliest study by author shows that the dichloromethane extractive(DEVJ)from the root and rhizome of Valeriana jatamansi Jones have an effect of antiproliferative on a panel of human cancer cellsTo further illustrate the substance foundations of anti-tumor effect of Valeriana jatamansi Jones,this work aimed to separate and purify an active fraction from dichloromethane extractive by Silica gel column chromatograph(CC)according to anticancer experiment in vitro.Then we analyze the the active fraction by the methods of TLC andUPLC-ESI-MS,aimed to illustrate its chemical component and the chemical structure typeSystematically study the chemical components contained in purified active fraction by MPLC、HPLC、PTLC and other modern chromatographic technique,and to analyze chemical structures of the compounds by 1HNMR、13CNMR、HR-ESI-MS、ESI-MS and other modern spectroscopic methodsMethods.1.The percolation method was used to extract the medicinal materials of Valeriana jatamansi Jones,then further extract the extraction by dichloromethane,and acquired further extract of the medicial material2.The method of silica gel column chromatograph(CC)was used to isoloate and purify the crude extract,acquired several active fractions3.The methods of TLC and UPLC-EST-MS were used to analyze chemical compounds of active fractions4.Using MTT assay to evaluate the anti-tumor effect in vitro5.The methods mainly including MTLC preparative thin layer chromatography(PTLC),preparative high performance liquid chromatography(HPLC),and other modern chromatographic technologies were used to isolate and purify compounds from active fractions6.Using modern spectroscopic methods including 1HNMR、13CNMR、HR-ESI-MS to analyze chemical structures of the compounds isolated and purified from active fractions7.Microculoure tetrazolium(MTT)assay was used to measure the antiproliferative effect of compounds on several humanized tumor cells,especially to investigate the dose-response relationship of the senstive tumor strainsResults.1.Three fractions named DEVJA、DEVJB、DEVJC have been separated from the crude extract(DEVJ)by the method of sillica gel column chromatograph(CC)2.The results of TLC assay showed that iridoids esters were the main constitute of DEVJB3.The method of UPLC-ESI-MS with a gradient elution was used to analyzed DEVJB,acetonitrile to water:acetonitrile was used as eluant.The result showed that there are several iridoid esters contained in DEVJB4.Anti-tumor experiment of DEVJB had been studied in vitro.The results showed that DEVJB had significant inhibitory effect on a panel of human cancer cells in vitro,such as SGC-7901、MKN-45、NCI-H1975、BXPC-3、HT-29.IC50,respectively were:5.94±0.13,1.39±0.07,2.61±0.10,1.71±0.06,,4.43±0.12μg/ml5.After separation,we acquired 5 compounds from DEVJB,and identified their chemical structure.There are 4 iridoid esters and 1 sesquiterpene lactone compound.The four iridoid esters are:Valtrate(1);Desoxidodrvaltrate(2);T-acetoxyl-Nardostachin(3);11-ethoxyl-pathchouli aleohol(4).The sesquiterpene lactone is 8-acetoxyl-pathchouli aleohol.They are all known compounds,8-acetoxyl-pathchouli aleohol was isolated from this plant for the first time6.The antitumor effect of the four iridoid esters had been studied in vitro.The results showed that the iridoid esters had significant inhibitory effect on the human cancer cell of MDA-MB-231.The IC50 was:19.02±0.25、23.49±0.36、21.03±0.18、49.71±0.41 μmol/L.8-acetoxyl-pathchouli alcohol have no antiproliferative effect on MDA-MB-231Conclusions.For the extraction and separation of Valeriana jatamansi Jones,we acquied crude dichlormethane extracts.DEVJB is an active fraction isolated and purified from the crude dichlormethane extracts by the method of sillica gel column chromatograph(CC).Our studies indicated DEVJB had significant inhibitory effect on five panels of human cancer cells in vitro.The main chemical compound of DEVJB are iridoid esters.We have separated 4 iridoids and 1 sesquiterpene lactone from it,and identify their chemical strucures.The four iridoids also have significant anti-tumor effect in vitroAll results in this study suggests that the primary efficacy material basis of DEVJB are iridoids,they all have antitumor effects.It has bright application prospect in finding leading antitumor drugs in iridoids.DEVJB has potential antitumor application values to tumor treatment. |
