| Compound proscription is the main form of clinical Traditional Chinese Medicine(TCM).The efficacy superiority of TCM is mainly carried out by compound proscription.By using modern methods we can reveal chemical and efficacy substance and physiological disposition of TCM,and set up a suit of consummate scientific quality control method,so that we can promote the development of science of Chinese materia medica,modernize the TCM industry and improve the international competitiveness of TCM.In this dissertation,HPLC and HPLC-MS technology were used in the research of two compound proscription-- Compound Ku-shen Injection and Compound Shi-Wei Tablets.Compound Ku-shen Injection is consisted of Sophoro Flavescens Ait and rhizome heterosmilacis,which has exactly inhibitory action on various cancers, prominent efficacy in alleviating pain and improving patients′quality of life.In this study,a HPLC method was developed to determine the content of 4 alkaloids(matrine,sophoridine,oxymatrine and oxysopho- carpine) in compound ku-shen injection,which has definite reference meaning in quality control.HPLC-MSn technology was used to analysis chemical components in compound ku-shen injection. Prior to the HPLC/ESI-MSn analysis, the studies on the fragmentation behavior of authentic chemicals of 4 kushen alkaloids were performed to find out the fragmentation pattern of the compounds. On this basis, a total of 13 kushen alkaloids, along with two flavones were identified in compound ku-shen injection.One of the flavones has never been separated from Sophoro Flavescens Ait and rhizome heterosmilacis by now.This result would be helpful in revealing chemical and efficacy substance of compound ku-shen injection. In the research of excretion components in rabbit urine after i.v.totally 18 components were identified,including 6 original ingredient from the injection and 12 metabolites, mainly glucuronide conjugate. By comparing the constituents in urine of different time,it can be concluded that original ingredients from this injection were excreted since 2 hour,abundant in 8 hour with little metabolites,and less original ingredients with lots of metabolites after 8 hour,which indicates that most of the original ingredients has been excreted in 8 hours.Abundant metabolites can be found within 18 hour urine,nothing can be identified after 18h.Metabolites were also identified in human urine after taking compound kushen injection orally. This study would be helpful in clinic medication and further exploitation of this preparation.To quantificationally study the physiological disposition of compound ku-shen injection, a HPLC method was set up in determination of sophoridine in rabbitsˊ plasma. Sophoridine in compound kushen injection was adequately described by a two compartment model,t1/2 was 92.67min,a little longer than the result in reference,which probably resulted in different compatibility and different administration Route. Compound shi-wei tablets transformed from shi-wei powder, recorded in zheng-zhi-hui-bu—an classic medicinal literature in ancient China. The tablets consists of four crude herbs: shiwei(Pyrrosia petiolosa (Christ) Ching, ). kushen(Sophora flavescens Ait.) ,bianxu(Polygonum aviculare L.) and huangqi(Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.)Hsiao).It is used in clinic for nephritis , chronic nephritis, pyelonephritis,diuresis.and so on.First,the content of matrine,sophoridine was determined by HPLC.In order to exactly identify the flavonoids in compound shi-wei tablets,a short review of currently available mass spectrometric methodology used in the structure elucidation of flavonoids is presented.30 components were identified from compound shi-wei tablets,firstly expatiate the chemical and efficacy substance of compound shiwei tablets.In the study of Serum Pharmacochemistry,6 compounds were identified, which maybe parts of the main efficacy substances of compound shi-wei tablets.3 multi-creatinine conjugates were also found in serum.which were also identified in urine. Creatinine is one of the important index in examining kidney function. Kushen alkaloids can be conjugated with multi cretinine and excreted,which maybe affect the result of determining creatinine in judge of kidney function.In the research of excretion components in rabbit urine after Po. compound shi-wei tablets,it can be concluded that the metabolits are mainly of the same type with the metabolits of Compound Ku-shen Injection,but the excluding time are different.Through the research of content experiment and the study of Serum Pharmacochemistry,it can be concluded that sophoridine is one of the active components in compound shi-wei tablets and should be determined in quality control.This study would be helpful in both revealing the chemical substance and improving the quality control of compound shi-wei tablets.Above all, the main efficacy substance of compound ku-shen injection are kushen alkaloids, concluding oxymatrine, N-oxysophocarpine, matrine, sophoridine, sophocarpine, N-methylcytisine, 5α,9α-dihydroxymatrine, baptifoline, hydroxyoxysophocarpine, and so on; the main efficacy substance of compound shi-wei tablets are kushen alkaloids and some flavones, conclude oxymatrine, N-oxysophocarpine, matrine, sophoridine, sophocarpine, baptifoline, hydroxyoxysophocarpine, calycosin, formononetin and so on.In conclusion,this study firstly revealed efficacy substance of the two compounds,elucidated the metabolites and metabolic pathways and provided the academic evidences for further research and clinical application. |
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