The Standard Research On The Quality Standard Of Phertima Dispensing Granules & Earthworm Gel Preparation

Objective: In order to control the quality of Phertima dispensing granule and assure its clinical results, System study on quality standard of Phertima material and its dispensing granule was carried out according to the theories of Traditional Chinese Medicine and"the Technical Requirement of Research on the Quality Standard of Chinese Medicine Granule"and the validity of hypoxanthine and succinic acid as the active components was testified, In addition, earthworm gel preparation was researched, and its therapeutic efficacy of scald was evaluated.Methods: This research includes quality standard of Phertima and its dispensing granule, preparation technology, pharmacodynamics evaluation of hypoxanthine and succinic acid, stability, earthworm gel preparation, pharmacodynamic action of scald. In the quality research of the raw materials, thin layer chromatography (TLC) was used to identify amino acids, succinic acid and extractive of trichloromethane. High performace liquid chromatography (HPLC) method used for determing the content of hypoxanthine using chromatographic column of Inertsil ODS-EP, mobile phase of H2O-CH3OH-C4H8O (100﹕0.5﹕0.5), flow rate of 1.0mL/min and detection wavelength of 254 nm. Another component, succinic acid, was determined by High Performance Capillary Electrophoresis (HPCE). The succinic acid could be separated and detected by buffer (0.36 mmol/L Tris-1.25 mmol/L H3BO3-0.5% 4-ethylene -5-amino-0.025 mmol/L CTAB adjusted to pH = 11.45 ) at -10kV of constant voltage. Other determination indexes, for example, dopant, moisture content, extractive assaying, also were determined.The dispensing granules were preparated according to traditional decoction, removing impurity by centrifuge, drying by spray and granulation and the extraction technics of active components were optimized by orthogonal design using the content of hypoxanthine as the valuation index. Same methods including TLC, HPLC which were used in the raw materials were applied to control quality of Phertima dispensing granule, other indexes were ascertained according to requirement of granules in appendix of Pharmacopoeia and preliminary experiments on stability were carried out.Asthmatic model of mice induced by histamine and acetylcholine and cough model of mice induced by ammonia water, were used to evaluate the pharmacodynamics of extract, dispensing granules, hypoxanthine, succinic acid and so on.Earthworm gel is composed of earthworm extract, carbomer, glycerine, propanediol and so on and its scald pharmacodynamics evaluated by scald area, vary of hydroxyproline and pathology slice.Results: In the quality research of the raw materials, TLC in China Pharmacopeia (version 2005) was improved, 2-aminopropionic acid and succinic acid, were added as the marker substance. The content range of hypoxanthine in different herbal Pheretima aspergillum were 0.54 mg/g～1.25 mg/g and 1.6 mg/bag~3.3 mg/bag in is dispensing granule by HPLC. The content of succinic acid was 0.8 mg/g in herbal Pheretima aspergillum (region:GuangXi).The optimum extract processing of active components was: 30 gram Phertima was weighed and soaked 1hour, then bored 1h and extracted with water third, used 12 volumes of water, afterward, filtrated, concentrated, dried and granulated. Dispensing granules were taken stability examination in the laboratory for three months including the outward appearance, the discrimination, the inspection, the assaying and the hygieology and all above were conformed to the requirement.Hypoxanthine and succinic acid have effects on cough-remedy, asthma-smooth. It can be concluded that the two ingredients as the standard components are reasonable and reliable. In the scald-remedy experiment, the data indicated that both earthworm syrup gel and earthworm non-syrup gel have therapeutics effects.Conclusions: The all methods to control the quality both the raw materials and the granules were standard and scientific and hypoxanthine and succinic acid as the standard components was reasonable. This thesis provided scientific and reliable basis for controlling the quality of Phertima dispensing granules. In addition, the earthworm gel preparation in agglutinating and the vertification of pharmacodynamics will be established an experimental foundation in Phertima comprehend research and development.