Injection Naomai Clear

Naomaiqing sterile powder for injection is compound preparation which composes of salvia miltiorrhiza,Panax notoginseng and borneol,mainly treat acute cerebral infarction and sequels. The formular originated from the People's Republic of China Pharmacopoeia 2005 version of Compound Danshen Tablet.On the basis of it,second development was carried out,with the help of macroresin refining and purification processes,preparation of the danshen intermediate. Combination of freeze-drying technology,studies including freeze-drying on forming process, stability of borneol during freeze-drying process,injection of quality standards and safety were carried out,successfully developed naomaiqing sterile powder for injection,in accordance with technical requirement of TCM injection research,improving the preparation of safety, effectiveness and stability.Through the plasma concentration,seven components pharmacokinetics studies were conducted in the in rats,and compared between single and compound preparation,conducted a much more comprehensive preliminary study,laid the foundation for clinical application.At first,TLC was selected for the qualitative analysis of index components,including danshen water-soluble components(SAB,RA,DSS,PA),sanqi mainly saponins(R₁,Rg₁,Rb₁) and bomeol.HPLC was selected for the quantitative analysis of index components,including SAB,RA,R₁,Rg₁,Re,Rb₁,Rd.GC was selected for the quantitative analysis of borneol and was used for organic residue inspection of danshen and sanqi intermediats.Established analysis method was accurate for the qualitative and quantitative analysis of the preparation.Adopt macroresin purification technology,danshen extract was purified.To select salvianolic acid B as index,using single-factor study,the final process was determined as follow: adjust danshen extract solution to pH 2(salvianolic acid B 4.06 mg·mL^-1),according to the ratio of salvianolic acid B and macroresin 26 mg·g^-1at the velocity of 1 mL·min^-1,adorption 30～40min, and then eluted with pH 2 water 5BV,20%pH 2 ethanol 4BV,respectively,eluted with 40% ethanol 5BV,concentrated and freeze-drying,danshen intermediate was obtained.With three batchs of danshen intermediates,the transfer rate of SAB were above 80%and the content of SAB were above 60%,make the active ingredient in the product much more reproducible,laied foundation for the futher study of the preparation,benefit to the stability of the preparation. Through studies of the type and amount of solubilizing agent to borneol,lyoprotectent, activated carbon and pH,selected the appearance and the content of borneol of freeze-dried sample as index,conducted formulation screening.The final formulation was determined as follow:danshen intermediate 45rag,PNS 15mg,borneol 6rag,HP-β-CD 108mg,Vc 2.5mg, mannitol 200mg.The eutectic point was -2.6℃.The freeze-drying process was as follow: pre-freezing temperature:ultra-low temperature refrigerator(-70℃)pre-freeze 3h,-35℃,0.5 h; 1st drying temperature:-10℃,12 h;2nd drying temperature:20℃,2 h.According to above formulation and preparation,five batches of samples were prepared,and in accordance with established analysis method for quality research.Results show that each bottle containing salvianolic acid B,rosemary acid were above 27.0rag,1.7mg,respectively,account for above 64% in danshen intermediate;notoginsenoside R₁,ginsenoside Rg₁,Re,Rb₁ and Rd were above 1.2mg,5.1mg,0.6mg,4.3mg,1.0mg,respectively,account for above 80%in PNS;borneol was above 5.8mg.Meanwhile,the studies of products safety(irritant,haemolytic,allergic)and acute toxicity were carried out.The results showed that in the formulation and preparation conditions, the active ingredients of finished products were stable,reproducible,have no allergic reaction on the guinea pigs,no rabbit red blood cells in vitro hemolytic,no obvious irritability on rabbit ear vein,no local irritation on intramuscular administration of the rabbits.Application of plasma concentration method to pharmacokinetic studies,compared single with compound preparation in rats,using UPLC-MS determination plasma,used DAS2.0 non-compartment model calculate plasma concentration.AUC_(0-∞)and t_1/2of salvianolic acid B, rosemary acid,notoginsenoside R₁,ginsenoside Rg₁,Re,Rb₁ and Rd in single preparation(danshen intermediate and PNS)were 5240±3212,381.1±138.2,2557±863, 5108±1291,461.54±141.3,362583±94805,69725±11614μg·h·L^-1,2.574±1.44,0.594±0.08, 0.68±0.33,0.494±0.17,0.384±0.10,15.154±3.18,16.73±4.97 h;corresponding datas in compound preparation were 57324±2622,551.94±186.2,25504±2402,5116±4369,591.64±549.6, 275417±33062,584694±12035μg·h·L^-1,4.034±1.10,0.81±0.24,0.714±0.41,0.904±0.56,0.394±0.11, 14.934±2.05,25.51±10.78 h.The results showed that pharmacokinetic parameters of single and compound preparation no significant difference(P＞0.05),it was initially implied that other components has no effect on SAB,RA,R₁,Rg₁,Re,Rb₁,Rd pharmacokinetics.It was laid the foundation for the compound preparation of pharmacokinetics study,and possessed the guided significance in clinical safety application.