The Pharmaceutical Research Of Wubie Granules

Premature Ovarian Failure(POF) is a disease caused by some factor,it characteristiced with amenorrhea,infertility,estrogen anhormonia and high GTH in those women befor the age of 40.It is a state that ovarian gonadal hormone hypofrontality or disability decreases the negative feedback to hypothalamic-pituitary axis,then leads the increase of GTH and the decrease of estrogen.The main feature of POF is disabilities of ovary,reproduction and endocrine.There are no effective therapies for POF recently,Western medicine have some serious side-effects,few of them can improve or even recover ovarian functions by hormone replacement therapy or immunodepression,There are still lack of desirable therapies for most patients.Traditional Chinese Medicine have its unique advantages in therapies for POF.The prescription of Wubie granules is a proved effective recipe that has been used for many years in the treatment for POF by the famous doctor of Traditional Chinese Medicine——Xia Gui-cheng.The recipe is made up of Radix Polygoni Multiflori Preparata,Rhizoma Atractylodis Macrocephalae,Radix Dipsaci,Fructus Lycii,Poria,Carapax Trionycis and Testis et Penis Canitis.This project is to study related literature,preparation technology,quality standard and stability of Wubie granules according to the demands of new preparations for Traditional Chinese Medicine.Literature Section:Each medicine in the recipe has been summarized,including development in source,characteristics,chemical constituent,qualitation and quantitative analysis and pharmacologic actions.Preparation Section:1.According to the theory of Traditional Chinese Medicine and the physicochemical property. of the active ingredient, preparation process has been designed and evaluated by pharmacodynamics indexes; 2.The optimum extraction process of Testis et Penis Canitis evaluated by pharmacodynamics indexes were as follows:extracting 2 times, using 10-fold of 95% alcohol amount for 1 h each time; 3.For further research, the optimum parameters of the supercritical CO2 fluid extraction of Testis et Penis Canitis and chemical constituents of extraction have been studied by GC-MS:the extraction pressure was 25MPa and temperature was at 65℃,1mL/g of 95% alcohol as entrainer, extracted for 2 h and the extractive Rate was 27.4%;44 types of fatty acids were identified from the extraction. Meanwhile,3 types of Testis et Penis Canitis extraction have been evaluated by pharmacodynamics and sex hormone was determined. The results are:the alcohol extraction and the SFE- CO2 extraction are superior to the water extraction. The concentration of TESTO and E2 in the 3 types of extraction are proximate, the SFE- CO2 extraction is highest,and the alcohol extraction followed; 4. Orthogonal design was used to optimize extracting process of Fructus Lycii,Poria and Carapax Trionycis,using the content of polysaccharide and nitrogen as indexes,and the results are: extracting 3 times, using 8-fold of water amount for 60 minutes each time; 5. Single factor and orthogonal design were used to optimize extracting process of Radix Polygoni Multiflori Preparata,Rhizoma Atractylodis Macrocephalae and Radix Dipsaci,The content of tetrahydroxystilbene-glucoside and asperosaponin VI were indexes. and the results are:Radix Polygoni Multiflori Preparata is shattered to coarse powder,then extracting 3 times with the others, using 8-fold of water amount for 60 minutes each time;Then orthogonal design was used to optimize the alcohol precipitation process of the water extraction,The results are:the relative density of the extraction is 1.15(determined under 60℃), adding alcohol to the extraction and making it to 60% alcohol solution, then letting it stand for 12 hours at room temperature; 6. Concentration and drying process were studied, The results are:the extraction is concentrated with 70-75℃in temperature,and the vacuum is 0.05-0.08 MPa;the temperature is 70℃,and the vacuum is 0.08-0.10MPa while drying; 7. Preparation process for the granules has been optimized; 8.Pilot scale experiments have been carried out using the studied preparation process.Quality Standard Section:1. According to the Chinese Pharmacopoeia (edition 2010) general principles for granules,the pilot scale products have been checked; 2. The identification methods of the medicines in the preparation have been established using TLC; 3. The simultaneous Determination method of tetrahydroxystilbene-glucoside and asperosaponin VI has been established using HPLC; 4. The method of finger print for the preparation has been established using HPLC.Stability Section:The accelerated test of the preparation has been carried out for 3 months,and the results show that it is stable in the test period.