Component Analysis And Activity Detection Of SLPE On Antioxidation And Antitumour Activity

Firstly, the experimental material which is Suillus luteus fruit bodies were extracted by Petroleum ether（PE）.After that, the PE extracts of Suillus luteus（SLPE） were purified by Silica gel column chromatography and Sephadex LH-20 gel column chromatography with different elution requirement repeatedly. At the same time, using different detection methods to analysis whether the solution fractions was a single component. Secondly, the structure of single components were identified by nuclear magnetic resonance spectroscopy（NMR）and mass spectrometry（MS） methods. And then, the antioxidant and antitumor activity were determined by using 1,1-diphenyl-2-picrylhydrazyl （DPPH） radical-scavenging capacity and MTT methods separtely.The results were as follows:1、Using different detection methods to analysis whether the solution fraction was pure when the PE extracts of Suillus luteus （SLPE） were purified by Silica gel column chromatography and Sephadex LH-20 gel column chromatography repeatedly. After this, Preliminary appraisal for five single components, named ns41、ns101、ns221、ns5 and ns7 separately.2、The structures of five single components were identified by NMR and MS methods. The results were :the ns41 was oleic acid; the ns101 was linolie acid; the ns5 was ergosterol; the ns7 was 3,6 - dihydroxy - （（2E, 6E, 10E） -3,7,11,15 - hexamethyl sixteen carbon -2,6,10,14 - tetraene） ethyl benzene, in addition, it was isomeride of suillin; but the ns221 was not pure that was analyzed by NMR, it may be a mixture of two kinds of aliphatic acid.3、Acting Vc as the positive control, four single components and some preliminary separation components were analyzed for their antioxidant activities through DPPH free radical-scavenging method, the result showed that all of the single component exhibited DPPH radical-scavenging activity in different degrees with a dose-dependence manner. And their antioxidant capacity in order was ns7>ns5>ns101>ns41. The single component ns7 had the strongest antioxidant activity against DPPH radical than any other all single components And preliminary separation components, its DPPH free radicals scavenging capacity had already reached 93.68% at the lowest concentration 0.5mg/mL, and also, the IC₅₀ value of single component ns7 against DPPH free radicals was 0.0044 mg/ml, which was significantly higher than that of positive control Vc（0.016 mg/mL）. This proved that ns7 has the very strong antioxidant activity. The IC₅₀ value of single component ns5 was 1.0348 mg/mL, and its antioxidant activity was also very strong. Preliminary separation components also had DPPH free radical-scavenging activity in different degrees on a dose -dependence manner, their order was ns0>ns6>ns1>>ns4-2>ns4>ns3>ns2>ns221.4、Antitumour activity of four single components and preliminary separation components were detected by MTT antitumour assay. The experimental research had suggested that single component ns7 had the highest antitumour activity with a IC₅₀ value of 7.47μg/mL in vitro, then was the single component ns5 and its IC₅₀ value was 108.54μg/mL, antitumour activity of single component ns41 and ns101 were relatively weak. The result had proved that single component ns7 could significant suppression of the proliferation of Human gastric carcinoma cell lines （BGC-823 tumor cells） in a dose-dependence manner. The preliminary separation components also had antitumour activity, among them the strongest of proliferation inhibition was ns3 with an IC₅₀ value of 95.77μg/mL, then was ns42、ns4、ns6, and their IC₅₀ value was 268.65μg/mL、296.62μg/mL、430.66μg/mL, respectively. Antitumour activity of other components were very weak with an IC₅₀ value of over 800mg/mL.The above test had indicated that all of the four single component that purified from Petroleum ether extracts of Suillus luteus（SLPE） had antioxidant and antitumour activity in different degrees, among them the ns7 revealed the strongest activity both antioxidant activity and antitumour activity. Single component ns7 belongs to polyisoprene phenol class compounds, which was an new structure compound and was first isolated from the mushroom Suillus luteus, in addition, its antioxidant and antitumour activity was not researched both at home and abroad.