The Research On Technics Rationality Evaluation Methods Of New Preparation Of Classic Xiaoyao Prescription

1.Objective:Using SFE-CO₂,separation of macroporous adsorption resin and other new technologies,new preparations were prepared.By comparing the chemical constituents and pharmacological effects and spleen correlation spectral efficiency,three preparation for Xiaoyao preparation traditional decoction,pills,new formulation,we build the chemical constituents and biological effects of the whole process of fingerprint characteristics,with reasonable evaluation method for unfettered,modern Chinese native medicine preparation for classic prescription rationality assessment technique provides a new idea.2.Methods We use SFE-CO₂,separation of macroporous adsorption resin and other new technologies,by GC-MS determine fat-soluble components extraction technology,use the orthogonal experiment design optimization method of water-soluble ingredients,separation and purification unfettered new preparation conditions;the best Based on new products,the traditional one xiaoyao pill three preparation,traditional thin layer of identification,the content and the fingerprint,establish quality control method,Using stomach "syndrome" animal models and spleen "disease" animal model,through the gastrointestinal function and neurotransmitters indexes,such as observation,compared to unfettered square three preparation stomach model nerve - endocrine - and the effect of gastric bowel function,Using fingerprint technology,three kinds of preparation for unfettered square internal and external mutual chemical composition,the application of grey relational fingerprint characteristics, research spectrum analysis method,this paper discusses the effect of correlation material foundation,and the effects of modern Chinese native medicine preparation unfettered square evaluation of extraction and separation technology, based on the rationality of the route with overall biological effects of chemical composition fingerprint characteristics of rationality assessment method.3.Results The optimum parameters of unfettered dongfang preparations for (1):the SFE - CO₂ extraction temperature,pressure 30Mpa about 50℃, analytical stress,temperature 8Mpa about 40℃,Entrainment preparation anhydrous alcohol consumption for 0.1 ml/g,Circular extractors 2h,The CO₂ flow 25～30L/h.(2) of macroporous adsorption resin purification:adopt D-101 resin,will adjust for 5～solution concentration 10mg/ml,column 2～3BV velocity/h.2BV distilled water,with the velocity,with 70%alcohol 20%ethanol for elution preparation,adjust pH in dilute aqua ammonia to 8.0～9.0.In 2BV/h, collect 4BV velocity elution eluent.(3) the low temperature dynamic extract: extraction,first and second 12 times the amount of water extraction,the second with 2h 10 times of water extracted 1.5h.By orthogonal experiment on the decoction of alcohol process parameters optimization,namely the decoction to concentrate the relative density of about 60 degrees Celsius(1.10).(4) flocculation clarification,tuckahoe,angelica,limit the decoction to concentrate out:water(1:1),add 5%to 10%of chitosan(amount),shake at 40℃,water heating temperature 15min,after 3000rpm centrifugal(placed 2h,min-1,15 min),take supernatant.We establish new formulation unfettered 6 kinds of TLC identification,content of measurement methods of ferulic acid and paeoniflorin,and HPLC fingerprint.With the state pharmacopoeia commission issued "medicine fingerprint similarity calculation software",evaluation results are similarity in more than 0.9.Results show that the new preparation quality is stable and controllable.Establishing three HPLC fingerprint in and outside the body,including new preparation 19 10 characteristic peaks,pills,21 peaks and decoction 15 peaks, 12 peaks for three common preparations in vitro fingerprint.Plasma HPLC fingerprint includes health animal fingerprint and stomach model animals fingerprint,three kinds of new drug of animal model preparation has 11 characteristic peaks,pill has 13 peaks,decoction has 13 peaks,including 7 peaks for three preparation,Healthy animals' new preparation has 8 characteristic peaks,pill has 10,decoction has 13,5 peaks for three preparation. The preparation of three kinds of the chemical constituents were analyzed, including new preparation has 9 characteristic peaks,pills has 9,decoction has 7, including 3 peaks for three preparation.Results show that the new preparation unfettered effective composition content,reveal the three kinds of preparation of material base.Soothing liver and strengthening spleen test results show that the three kinds of preparation can improve the rat weight and plasma monoamine neural transmitter NE,5-HT concentration,make gastric tissue cells MTL gastrointestinal hormones,SS lower expression expression,Mice intestine motional rate reduced spleen,improve the thymus and spleen index,etc.The efficacy of the new is be equivalent to the decoction,and superior to such dose of pills.In vitro correlation spectral efficiency analysis,results of macroporous adsorption resin of chemical composition is t_R 27.803,29.726 peaks are highly associated to soothing liver and strengthening spleen function,t_R 46.438 peak of SFE-CO₂ extraction of chemical composition is highly associated to soothing liver and strengthening spleen function,which provides the index selection process.In vivo and in vitro correlation of three preparation in vitro correlation spectral efficiency in accordance with the size of the order is the new preparation peaks＞pills peaks＞decoction peaks. 4.Conclusions The rationality evaluation method of preparation technology based on the chemical constituents and biological effects of the whole process of fingerprint characteristics show that the technics of new preparation is reasonable,the quality is controllable,and the drug effect is reliable,which provides modern TCM preparation technology research a new idea and method.