The Study On Quality Control And Stability For The Innovative Demethylzeylasteral

Demethylzeylasteral was a monomer isolated from the traditional Chinese herbal medicine, tripterygium wilfordii Hook F. and lots of effects, immunosuppressive anti-inflammatory, anti-caner and antifertility was examined. Innovative drug demethylzeylasteral was extracted by the Pharmaceutical Department of Zhongshan Hospital affiliated Fudan University following a simple route, and was planned to apply for state class new medicine according to FDA requests.By elemental analysis, UV, IR, NMR (13C,1H) and mass spectrometry for structural confirmation of demethylzeylasteral, reliable raw material for the study on quality standards and stability was provided. The article was divided into the following two parts:the first part was titled the study on quality control and stability of the innovative drug demethylzeylasteral, the second part was titled determination of simultaneous determination, homocysteine, cysteine and cysteinylgycine by HPLC with fluorescence detector following2-cyano-6-aminobenzothiazole derivation.Part1:The study on quality control and stability of the innovative drug demethylzeylasteral1. The quality control of demethylzeylasteralThe quality standard of demethylzeylasteral was investigated according to Chinese Pharmacopoeia and technical specifications of the drug quality standards. Based on experimental results, such as the study of Sulfainib characterization, the detection of related substances, residual organic solvent and assay of demethylzeylasteral, this thesis established metods of indentify demethylzeylasteral by chemical reaction, UV and infrared spectrometry. The related substances were detected by HPLC. The method of detecting methanol, acetone, n-hexane, ethyl acetate was established by using GC and the assay of demethylzeylasteral was established by HPLC.2. The stability study of demethylzeylasteralStability of demethylzeylasteral was conducted according to the guiding principles of drug stability test, including impact factors test, accelerated test and long-term test. The sample characteristics, the main component content and related substances of stability samples were analyzed. It was found that demethylzeylasteral was sensitive to light, temperature and humidity. It should not be stored under the condition of high temperature, humility and strong light.The accelerated test indicated that demethylzeylasteral was stable at40℃, humidity of75%for6months. The six-months-long-term test indicated that demethylzeylasteral was stable at6℃, humidity of60%, too. The long-term test is still carrying.For statistical analysis on the validity, if main component content of demethylzeylasteral becomes98.5%, the retention time is21months. So the demethylzeylasteral is valid for18months.According to the stability study, demethylzeylasteral was suggested to be stored under the condition of low temperature, dark and dry.3. The draft quality standards of demethylzeylasteralThe draft quality standard of demethylzeylasteral was formulated according to the first two chapters of the research content. This chapter provides norms and basis for quality control of demethylzeylasteral.Part2:Simultaneously determination homocysteine, cysteine and cysteinylgycine by HPLC with fluorescence detector following2-cyano-6-aminobenzothiazole derivationA simple, sensitive, mild high performance liquid chromatography method with fluorescence detector was developed for simultaneous detection and quantitation of homocysteine (Hey), cysteine (Cys) and cysteinylgycine (Cys-Gly) in human plasma. The above amino acids were derivatized with a large excess of2-cyano-6-aminobenzothiazole (CBT), a thiols and primary amines-specific reagent, to an instantaneous and complete derivatization, prevented the form of disulfides with use of tris (2-carboxyethyl) phosphine hydrochloride (TECP). The sample was precipitated with acetonitrile-water60:40before being injected the chromatograph. The detection limits of Cys, Hey and Cys-Gly were2,5and1.5nM, respectively. Furthermore, the interference with glutathione (GSH) and metal ions was negligible.