| Chinese herbal formula is the specific performance of syndrome identifying treatment variation and holism concept. It is a complex system with the characteristics of complex chemical composition and many factors interfere with. Chinese herbal formula cures disease through concerted application of monarch, minister, assistant and guide,according to the disease pathogenesis, absorbed through the appropriate route after administration and cure diseases by overall effect.So the study of the chinese herbal formula should base on the overall concept.The basic research is one of the core issues of modernization of Chinese medicine, it may essentially clarify the role of the material foundation of the formula and provide the basis for optimizing the prescription, making scientific preparation process and improving the quality of preparation.Qing-Luo-Fang is a representative herbal formula used to treat Rheumatoid Arthritis. To reveal the material basis, variation of the drug in the body and improve the quality of Qing-Luo-Fang. In this thesis, sophisticated analytical techniques such as HPLC-DAD, HPLC-DAD-MS/MS were used to study and clarify the material foundation. This thesis studied the pharmacodynamic chemical of normal rats and pathological rats in Qing-Luo-Fang by the serum pharmacochemistry method. According to the results of pharmacodynamic chemical in vivo, a HPLC-DAD method for the quantitative determination of 8 main effective substances(included 5 compounds detected in serum) was established. The research ideas are as follows: The first part:summary of the related literatureDocuments about the clinical applications and modern researches of Qing-Luo-Fang and brief introduction of Sophora flavescens Ait., Sinomenium acutum, Phellodendron amurense Rupr. were summed up and analyzed, and the major constituents of Qing-Luo-Fang were concluded in the preface.56 references were cited. The second part:qualitative analysis of the main constituents of Qing-Luo-FangThe HPLC-DAD-MS/MS analysis method was established and the main constituents in Qing-Luo-Fang were identified by coupling technique HPLC-DAD-MS/MS firstly.14 alkaloids including magnoflorine, oxymatrine, sinomenine, matrine, berberine and 2 unknown constituents in Qing-Luo-Fang were elucidated. The type of 14 main alkaloids included:aza-cyclohexane alkaloids (matrine), aza-cyclohexane alkaloids (sparteine), high-benzyl tetrahydroisoquinoline alkaloids (tetrahydroprotoberberines), aporphine (aporphine quaternary ammonium alkaloids),tetrahydroisoquinoline alkaloids (prpaporphine). the main constituents in Qing-Luo-Fang were identified and the results lay a good foundation for further serum pharmacochemistry study and quantitative analysis study of the major substances in Qing-Luo-Fang.The third part:preliminary study of serum pharmacochemistry in normal and model ratsOn the basis of the established analysis method of HPLC-DAD-MS/MS, by animal model-making,optimizing the pretreatment conditon of plasma and the chromatographic method, the analysis method of serum pharmacochemistry for Qing-Luo-Fang was established. From a comprehensive analysis of the HPLC-DAD-MS/MS chromatography of Qing-Luo-Fang, plasma containing drug, blank plasma,13 chromatogram peaks were observed in normal rat's plasma. Among them,7 original compounds from Qing-Luo-Fang and three type of metabolites were identified.14 chromatogram peaks were observed in model rat's plasma. Among them,7 original compounds from Qing-Luo-Fang and three type of metabolites were identified.The commom original compounds from Qing-Luo-Fang of nomal and model rats were magnoflorine, lupanine, matrine, sinomenine, sophocarpine, berberine and an unknown constituent. The metabolites of normal rats and model rats contain three types:oxidation of hydroxyl metabolites, glucuronide products and an unknown type metabolite. Most of these identified compouds were reported to have related effect on Rheumatoid Arthritis disease. At the last part of the chapter, the difference of the constituents absorbed into blood between normal rats and model rats was analysted and elucidated. There is some distinction about the absorbing and metabolizing ability of normal rats and model rats. The investigation of the absorbed components in nomal and model rat plasma after oral administration of Qing-Luo-Fang in principle clarified its real effective substances in vivo.The forth part:quantitative determination of eight main constituents of Qing-Luo-FangAccording to the result above, a HPLC-DAD method for the quantitative determination of 8 main effective substances (included 5 compounds detected in serum) was established. Through simultaneous quantitative determination of major components in Qing-Luo-Fang, not only elevate the quality control standards of Qing-Luo-Fang but also make the clinic application of Qing-Luo-Fang more effective and reliable.Through the identification of the main constituents of Qing-Luo-Fang, serum pharmacochemistry study of nomal and model rats after oral administration, and quantitative determination of eight main constituents of Qing-Luo-Fang by using coupling technique such as HPLC-DAD-MS/MS and HPLC-DAD, the real effective substances were clarified. It provided references for further studies of pharmacokinetics and active mechanism research. The method was simple, accurate, quick and with a good reproducibility. This study achieved identifidation of all main constituents in Qing-Luo-Fang and in plasma of nomal and model rat, determination of main effective substances. It may lay the root for further study of quality control fingerprint and serum chemistry, and it can provide basis for clinical use and further development and utilization of Qing-Luo-Fang and made contribution for valuable research of modernization of Chinese medicine.
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