Studies On Preparation Process,Quality Standard And Pharmacodynamics Of Erhuang Granules

Objectives:The original prescription of Erhuang Granule,which derived from the old Chinese medicine doctor's experience prescription,is composed of Astragalus membranaceus,Trigonella,and foenum-graecum etc.Its efficacy is to support the kidney and tonify qi,activate blood and clear away turbidness.It was always used for the treatment of chronic renal failure?CRF?,improving renal function and alleviating the clinical symptom.We made decoction of Erhuang as granules,and studied its preparation process,quality standard,preliminary stability,pharmacodynamics and acute toxicity test systematically.The purpose of this topic is to finish preclincal studies and to provide a basis for further clinical trials according to the requirements of TCM hospital preparations.Methods:1.Preparation Process:Taking the water quantity,boiling frequency,and times as the investigating factors by L₉?3⁴?table of orthogonal test.Taking the dried cream yield,contents of astragaloside and trigonelline as indexes.These data were analyzed by comprehensive evaluation of multiple mark to optimize extraction Process.Types and dosage of the excipients were selected by investigated hygroscopicity,granulation,fluidity and bulk density to optimize the best formation technics of Erhuang granules.2.Quality Standards and Preliminary Stability:Qualitative identification of Astra galus membranaceus,Rheum palmatum and Smilax glabra in Erhuang granules were carried out by thin-layer chromatography?TLC?;The content of astragalos ide was determined by HPLC-ELSD;Under mon-section preparation general i nformation of the Chinese Pharmacopoeia?2015?,we checked the traits identifyc ation,microbial limits,moisture content,particle size,melting property and con tent uniformity of Erhuang granules through all item regulations of granule.W e investigate the preliminary stability about three types of Erhuang granules by classical stress test,accelerated test.The traits,identification,microbial limits,moisture content,particle size,melting property and content uniformity of Erhu ang granules wheather or not change,wich can provide the foundation for the s tability and the technological feasibility of granule,which can also optimaze th e preparation storage condition.3.Study of pharmacodynamics:Rats model of chronic renal failure was induced.HE stain was used to observe the pathological changes of tissues of kidney.Expressions of E-cadherin,?-catenin,FN and LN were observed by the metho d of immunohistochemical analysis.the maximum administration dosage?MAD?and acute oxity were determined on mice by oral gavage,and provide science fundation for the clinical administration and dosage.Results:1.The routes for preparing of Erhuang granules were:medical material were coarsed powder,and then boiled 1 hours with 10 times amount of water,boiled 3 times.The water decoctum was filtered,incorporated,centrifuged and concentrated with reducing pressure until dense is 1.1¹.2 by vacuum drying?80??.Then it is dried and crushed to powder and screened through mesh size of 80;The suit proportion of adjuvant were 2 powder and 1 dextrin,then mixed well.80 percent ethylalcohol was maked as wetting agent to make for soft wood and pellet,dring under 80 centidegree for 3 hours,mending and split charging.2.Establishment of quality standard and preliminary stability of Erhuang granules.Astragalus membraneceus,Rheum palmatum and Smilax glabra in Erhuang granules were identified respectively by TLC,The TLC spots were clear and specific in good repeatability without inference from negative sample.The contents of astragaloside was determined by HPLC,Linear ranges,precision,stability,repeatability,recoveries of astragaloside was well and linear ranges of astragaloside was 1.260²0.160?g?r=0.999 6?,with average recoveries of 96.82%?RSD 0.91%?respectively.Under the preliminary stability study process,we inspected respectively granules'trait,moisture,microbial limit,qualitation appreciation,and assay etc by classical stress test and accelerated test.The results were not marked change and every index consistented with specify.It was stability on the pot life.3.To make rat modle through adenine,expressions of?-catenin,FN and LN in rat renal tissue are elevated.but the expressions of E-cadherin were significant improved with treatment of Erhuang extract.In the experiment study of acute toxicity,Erhuang Granules is of low acute toxicity to mice,and the maximum administration dosage is16.86 g?kg^-1/d on mice,which is equivalent to 84.3 times of the clinical dose.Conclusion:The optimized preparation process is reasonable,stable,and feasible for industrial production;The method of control quality is simple,accurate,specific and can be used for the quality control of Erhuang granules.Results of rat chronical renal failure function impairement modle test show that the effection of improving chronic renal failure fuction of Erhuang Keli is related with index reversals.The acute toxicity experiment show that the clinical prescribed dose of the medicine is in a low toxicity..