| Herb couple (mixture of two kinds of herbs) is the basic composition unit of Chinese herbal formulas and has special clinical significance. Although herb couple is much simpler than complicated formulas in composition, it still retains the basic therapeutic features. It is the special form of complicated formulas.The mode of composing, status and function of herb couple are comprehensively discussed. The recent progress of physical and chemical changes during combination of Chinese traditional medicines is reviewed. Constituents in herbs are so complex that it is difficult to obtain a definitely conclusion on chemical reactions occurred during combination. To simplify investigation procedure, a new idea of studying combination of herbs was proposed: different constituents or similar compounds were extracted from each herb and then combined. Material basis of coptis-evodia, veratrum-paeonia and veratrum-ginseng herb couples were studied. The research work described in this dissertation includes extraction, purification, analysis and combination. (1) coptis-evodia herb coupleProcess of extracting total flavonoids from Fructus Evodiae[Juss] Benth was studied by spectrophotometric method. The optimal procedure of extracting total flavonoids was reflux for 4 hours with 50% ethanol as solvent. The yield of the total flavonoids was 33.6mg/g. The total flavonoids were identified by color reactions to be composed mainly of flavone and flavonol. The extract of Fructus Evodia was purified by solvent extraction, column chromatography and recrystallization. Hyperin and isorhamnetin-3-O-galactoside were identified by mass spectrometry (MS), !H nuclear magnetic resonance (]HNMR) and I3C nuclear magnetic resonance (13CNMR). A RP-HPLC method for simultaneous determination of hyperin and isorhamnetin-3-0-galactoside in Evodia rutaecarpa [Juss.] Benth was established. A Diamonsil-C18 column (5m 250mmx4.6mm i.d.) was used and a mixture of methanol-0.5%phosphoric acid (50:50) was used as mobile phase at a flow rate of 1 .OmL/min. The column temperature was 30 and the UV detection wavelengthIIIwas 360nm.Purification process of total alkaloids of Rhizoma Coptidis by using macro-porous resin chromatography was studied. It was found that D-101 was more effective for purifying total alkaloids compared with other macro-porous resins. By chromatography of D0101, the content of total alkaloid was enhanced to 83.7% from 37.7% with a recovery of 71.8%.The change of chemical constituents during combination of Rhizoma Coptidis Alkaloids with Fructus Evodiae Flavonoids was studied. Different proportions of Rhizoma Coptidis alkaloids and Fructus Evodiae flavonoids were combined and heated for 4h at 70. The solutions were analyzed by RP-HPLC and the fingerprints were compared. The results showed that only little changes of the relative peak area of different chemical constituents were observed and no new peaks were detected after combination.Rhizoma Coptidis and Fructus Evodiae were decocted separately, and then the combination of the two extracts was studied by mixing the extracts. By this way, the effect of one constituent on the solubility of another constituent and the absorption by the residue during decocting together was avoided. Rhizoma Coptidis and Fructus Evodiae were decocted separately with water. Different proportions of Rhizoma Coptidis aqueous extract and Fructus Evodiae aqueous extract were combined and heated for Ih at 100 The samples were analyzed by RP-HPLC and the fingerprints were compared. The results showed that chromatographic peaks were additive and no new peaks were detected. No obvious change of the relative peak area of main chemical constituents of Fructus Evodiae was observed. However, the relative peak area of main chemical constituents of Rhizoma Coptidis decreased linearly with the content of Fructus Evodiae aqueous extract. The solubility of main alkaloids in Rhizoma Coptidis decreased under the presence of constituents from Fructus Evodiae. (2) veratrum-paeonia herb coupleThe process conditions of isol...
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