Studies On The Optimization Of Prepar Ation Process And The Improvement Of Quality Standards For Heparolysate Injection

Heparolysate injection and Heparolysate for injection can promote proteinsynthesis and reduce the protein decomposition,Promote the proliferation andregeneration of normal liver cells.Having protective effect on liver injury induced bycarbon tetrachloride.Reducing glutamine-pyruvic transaminase,promote tissue’srecovery.Mainly used for auxiliary treatment of chronic hepatitis and cirrhosis of theliver.This topic in the original foundation of quality standard of Heparolysate inje-ction.The research is add to improve the quality standard of Heparolysate injectio-n.According to test results of pHarmacopoeia standards and improving standards,to optimize the process of one manufacturer.The purpose is to increase the safety and efficacy of Heparolysate injection.Using HPLC to confirm the chromatogram of polypeptide; Using UV-spectropHotometric method to confirm the sodium bisulfite contents of Heparolysate inje-ctions and definite the limes;Using cation exchanging chromatograpHic separation and post column derivatization to detect the contents of about free AA and hydr-olysis AA of Heparolysate injections,and to determine the limits of hydrolysisAA minus free AA; The biological safety test method specified in the pHarmacopo-eia were detected on the safety of Heparolysate injections;Using MTT,CCK-8met-hods to detect the activity of polypeptide injection;Using Gel ChromatograpHy to confirm the molecular weight distribution.According to test results of pHarmacopoeia standards and improving standard-s,on the basis of manufacture’s process,using polypeptide yield and activity asan index, to optimize the extraction process.Single factor test include enzyme boilin-g time, solution before the hydrolysis time, temperature, pH and trypsin concentr-ation effects on peptide yield.On the basis of single factor test,the orthogonaltest was adopted to optimize the peptide enzymatic hydrolysis conditions.To determi-ne the optimal extraction process is:before enzyme boiling15min,hydrolysis time3H, temperature45℃,pH8.5,enzyme concentration6.0g/kg.pHarmacological experiments indicate: according to the optimization of extrac-tion process,not only the content of polypeptide but also the peptide activity inc-reased significantly.Based on the research of this standard, there are obviously i-mprovement in the two aspects of Heparolysate injection’s safety and efficacy.Ac-hieving the purpose and requirements of this research.