Research On The Chemical Constituents And Quality Control Of Herba Sedi (Sedum Sarmentosum Bunge)

1.Objective: Chuipencao (Herba Sedi) originates from the fresh or dry herb of Sedum sarmentosum Bunge, which exists extensively, being widely used as a folk TCM for kinds of hepatitis and other diseases. Its formal name is Gouyabanzhi, recorded by the book "Compendium of Materia medica". People consider it cool and sweet, then apply it to clearing heat, detoxifying, excreting dampness. In addition, in clinic , it can cure suelling, gynogenic infection, ulceration of skin, jaundice due to damp-heat, difficult dysuria and less urine as well. So, Herba Sedi is the main component of Chuipencao powder preparation, which is to be taken after being infused in boiling water. However, it's reported that its glycosides is not suitable to be as a contrast because of its instability when meeting hotness, acid and alkaline compounds. On the basis of the research on active constituents, the purpose of our study is to find out a contrast to be as a norm controlling the quality.2. Method: Our studies included six parts as follows: (1) Herbal verification and pharmacognosy study of Herba Sedi. (2) Study on chemical constituents. Extract with 95% ethanol 3 times, recycle ethanol, and after being suspended with water, extract with chloroform, ethyl acetate and n-butanol, isolate chemical constituents from these extractions with silica gel column, polymide column, Sephadex LH-20 column, and C18 column, and lastly identify them with UV, IR, 1HNMR, 13CNMR. Using the method of activity track and the result of activity sifting, we made a system isolation study on the chemical constituents of Herba Sedi. (3) Discussion about pharmacological activity measure. According to the activities of the herb, effects of the compounds on the transformation and proliferation of T,B lymphocytes of mice spleen, and both on the ALT content in the culture media of liver cell injured by CCl4 were studied in vitro and in-vitro. All these work is to define the effective compound. (4) Contrast research. Define the contrast on the basis of the specialization of constituents and the result of sifting. Also, do research on the quality and stability of contrasts. (5) Research on the methods of qualitative analysis and quantitative analysis with TLC and HPLC. (6) Preferably choosing HPLC, make a discussion on the fingerprint graph of Herba Sedi.3. Result: By means of those works, we have made a progress on quality control of Herba Sedi, and 31 compounds were isolated from chloroform extract, ethyl acetate extract and n-butanol extract, 20 of which were defined by spectrum analysis and chemical analysis:tricin-7-O-β-D-glucoside(Ⅰ),Luteolin(Ⅱ),Isorhamnetin (Ⅲ),Isoliquiritigenin (Ⅳ),δ-Amyrione (Ⅴ),3β,6β-Stigmast-4,22-dien-3,6-diol(Ⅵ),quercetin (Ⅶ),β-Sitosterol (Ⅷ),Daucosterol (Ⅸ),pentadecyl mercaptan (Ⅹ), 4-hydroxy-3,5-dimethoxybenzoic acid (Ⅺ),3,4,5-trimethoxybenzoic acid (Ⅻ),dioctadecyl sulfide (ⅩⅢ),palmic acid (ⅩⅣ),benzoicac acid methyl ester (ⅩⅤ),3-epi-Amyrin(ⅩⅥ),δ-Amyrin(ⅩⅦ),beta-D-galactopyranosyl-(3-1′)-beta-D-glucopyranosyl-(4′-1)-beta-D-Allose ⅩⅧ ,KNO3 ( compound A ) and NaNO3 ( compound B ). Among them, compound Ⅵ,Ⅶ,Ⅹ,Ⅻ,ⅩⅢ,ⅩⅣ,ⅩⅤandⅩⅧ were discovered in this herb for the first time, and other 9 compounds are still being identified. Furthermore, in-vitro activity experiment showed that compound Ⅰ can obviously restrain the transformation and proliferation of T ,B lymphocytes cell in mice spleen, injured by CCl4. Also, the ethanol extract of Herba Sedi had certain effect, and so does ethyl acetate extract, n-butanol extract, and water extract. Depending on the experiment of index constituents, tricin-7-O-β-D-glucoside possesses its specialization and superior immunity-controlled effect, so that it could be an index. Besides, δ-Amyrin has the same effect as tricin-7-O-β-D-glucoside,so it may become another index. Study and optimize the making process of tricin-7-O-β-D-glucoside, and believe that its productivity would be higher when extracted by 60% ethanol, direc...