Study On Constituents Absorbed Into Blood After Oral Administration Of Shao-yao Gan-cao Tang

Safety, effectiveness and quality control are the basic properties of modern medicine. A drug could be recognized by modern society, these three properties are indispensable. To develop Traditional Chinese Medicine into a modern medicine, we must solve the issue of material foundation of efficacy of Traditional Chinese Medicine. Traditional Chinese Medicine and the human body are two separately complex system, the interactions between two complex systems rely on chemical information to exchange. Studying the chemical information flow between the two complex systems is the entry points to resolve the discipline between the two complex systems, without interaction we could not study the material basis of efficacy of Traditional Chinese Medicine, so we study the transitional components absorbed into blood after oral administration. The thesis used Shao-yao Gan-cao Tang(SGT) as a drug tool to establish analysis method in vivo, isolate and identify components absorbed into blood. Preparing for the material basis of efficacy of Shao-yao Gan-cao Tang, using the target cells or target enzyme affinity binding to blood constituents, the ultimate goal is to build the method of research on material basis of efficacy of Traditional Chinese Medicine in vivo.1. Adscription of transitional constituents absorbed into bloodBy studying the chemical components of Shao-yao Gan-cao Tang in vivo and vitro, we established the HPLC fingerprint of Shao-yao Gan-cao Tang. On this basis, using the established methods, analyzing and comparing HPLC fingerprint spectrum between Shao-yao Gan-cao Tang and serum samples, according to the retention time of each component we determined the prototype transitional components absorbed into the blood after oral administration of Shao-yao Gan-cao Tang and their metabolites. By comparing with the HPLC fingerprint spectrum of Shao-yao Gan-cao Tang(SGT), Paeonia lactiflora extractions(S), Glycyrrhiza uralensis extractions(G), the control sample of single herb with rats serum samples of Paeonia lactiflora, Glycyrrhiza uralensis, Shao-yao Gan-cao Tang, according to the retention time of each component we determined the prototype components absorbed into the blood from source of medicine. The results show that there are 32 transitional components(SGT1-SGT32) absorbed into blood in serum samples after oral administration of Shao-yao Gan-cao Tang, including 17 components of the original parties absorbed into blood directly,15 components of the metabolites. After oral administration of Paeonia lactiflora decoction, there are 8 components(Sl-S8) absorbed into blood in serum samples, after oral administration of Glycyrrhiza uralensis decoction, there are 22 transitional components(G1-G22) absorbed into blood in serum samples. Transitional compounds SGT1, SGT2, SGT6, SGT7, SGT14, SGT16, SGT20, SGT26 absorbed into blood originated from the constituents S1-S8 absorbed into blood of Paeonia lactiflora, components SGT1, SGT2, SGT6, SGT7, SGT14 are directly original parties absorbed into blood after oral administration of Paeonia lactiflora decoction. Transitional compounds SGT3, SGT4, SGT5, SGT8, SGT9, SGT10, SGT11, SGT13, SGT15, SGT17, SGT18, SGT19, SGT21, SGT22, SGT24, SGT25, SGT27, SGT28, SGT29, SGT30, SGT31, SGT32 absorbed into blood originated from the constituents G1-G22, SGT3, SGT4, SGT8, SGT9, SGT10, SGT11, SGT15, SGT17, SGT18, SGT21, SGT25, SGT29 are directly original parties absorbed into blood after oral administration of Glycyrrhiza uralensis decoction. Compound SGT12 and SGT23 are the newly emerging substances absorbed into blood in rats serum after oral administration of Shao-yao Gan-cao Tang. Some plasma concentration of serum samples were obviously increased, such as SGT6, SGT7, SGT10, SGT11, SGT14, SGT15, SGT16, SGT17, SGT18, SGT24, SGT25, SGT26, SGT27, SGT28, SGT29, SGT32. The bioavailability were obviously enhanced after combination of Paeonia lactiflora and Glycyrrhiza uralensis, it shows synergism between Paeonia lactiflora and Glycyrrhiza uralensis. The results obtained the same conclusions compared with other methods, it certified that serum HPLC fingerprint can also be used for the compatibility of Chinese herbal formula.2. Isolation and identification of transitional constituents absorbed into bloodBy the guidance of serum HPLC fingerprint spectrum of Shao-yao Gan-cao Tang, combining traditional techniques with modern preparative chromatography separation methods(silica gel column chromatography, ODS, MDS, Sephadex LH-20, macroporous resin D101 and preparation phase), to analyze, isolate and identify the prototype components absorbed into the rats blood from Paeonia lactiflora and Glycyrrhiza uralensis.Classified Paeonia lactiflora aqueous extract by D101 macroporous resin, from 10% ethanol eluate,30% ethanol eluate and the distilled water eluate we isolated five monomer compounds, by comparing HPLC fingerprint spectrum of serum sample after oral administration of Shao-yao Gan-cao Tang with the control serum samples, we confirmed that transitional components absorbed into blood after oral administration of Shao-yao Gan-cao Tang are SGT1, SGT2, SGT6, SGT7, SGT14 from Paeonia lactiflora. Combination with ESI-MS,1H-NMR,13C-NMR data we determined that the prototype components absorbed into rats blood from Paeonia lactiflora, SGT1 was gallic acid, SGT2 was isopaeoniflorin, SGT6 was albiflorin, SGT7 was paeoniflorin, and SGT14 was isobenzoyl-paeoniflorin. Classified Glycyrrhiza uralensis aqueous extract by D101 macroporous resin, from 10% ethanol eluate,30% ethanol eluate and 50% ethanol eluate we isolated eight compounds, by comparing HPLC fingerprint spectrum of serum sample after oral administration of Shao-yao Gan-cao Tang with the control serum samples, we confirmed that transitional components absorbed into blood after oral administration of Shao-yao Gan-cao Tang are SGT8, SGT10, SGT11, SGT15, SGT17, SGT18, SGT21, SGT29 from Glycyrrhiza uralensis. Combination with HPLC, ESI-MS, 1H-NMR,13C-NMR data we determined the prototype components absorbed into rats blood from Glycyrrhiza uralensis, SGT8 was isoviolanthin, SGT10 was 4'-O-[β-D-apiofuranosyl (1→2)-β-D-glucopyranosyl] liquiritigenin, or liquiritigenin-apiosyl (1→2) glucoside, SGT11 was liquiritin, SGT15 was licuraside or 4-0-[β-D-apiofuranosyl (1→2)-β-D-glucopyranosyl] isoliquiritigenin, SGT17 was isoononin, SGT18 was isoliquiritin, SGT21 was liquiritigenin, SGT29 was Glycyrrhizic acid.Our studies showed that the prototype constituents absorbed into rats blood after oral administration of Shao-yao Gan-cao Tang were similar to the composition that reported material basis of efficacy of Shao-yao Gan-cao Tang, such as paeoniflorin, glycyrrhetinic acid, it indicated that constituents absorbed into the blood between many ingredients may be the active components. It is feasible to study the material basis of the efficacy of Traditional Chinese Medicine from the constituents absorbed into the blood. If we can use the target cells or target enzyme combining affinity constituents absorbed into blood, it will further confirm the material basis of efficacy of Chinese Medicine compounds, on the basis, establishing the quality control methods of material basis of efficacy of Traditional Chinese Medicine associated with the clinical activity, providing the new ideas and methods for the study of material basis of efficacy of Traditional Chinese Medicine and its compounds, providing the reference of ideas and methodology for the research modernization of Traditional Chinese Medicine.