| Purpose and significanceclimacteric syndrome is a syndrome which is coursed by function decline or loss of Female ovary.it can lead to the endocrine dyscrasia,low immunity,vegetative nervous disorder and Systemic multiple pathological change[1].The main clinical manifestations of climacteric syndrome are menstrual disorder,vexation,irascibility,palpitation,insomnia,feverish sensation in the palms and soles,sweating,edema.With the acceleration of population aging process,the increasing speed on the number of women who is entering climacteric is also acceleration.At present,there are about 10 million women who are entering climacteric in each year in our country.This figure will be 217 million in 2030,which will account 1/7 of total population[2].Menopausal is a physiological process which must be experienced during women’s lives.There is 25 percent women will feel uncomfortable which can effect their job and life quality[3].The prescription of Musu is an experience prescription to treat the Climacteric syndrome,which is consisted of Rehmannie glutinosa Libosch,Cornus officinalis Sieb.et Zucc,Dioscorea opposita Thunb,Lycium barbarum L,Perilla frutescens(L.)Britt and other six herbs.Musu prescription is a decoction which has a good effect to treat the Climacteric syndrome.But the decoction is inconvenient to carry,poor taste,easy moldy deterioration and the process of preparation is too difficult to control.In this study,we will base on the Traditional Chinese medicine theory and modern Chinese medicine technology to study the preparation process and quality standards of the Musu Granules,which according to the requirements of new drug registration.Methods1.Study on preparation of Musu GranulesThe compound of herbs in Musu can solute in waters.So all herbs of Musu prescription were extracted by water,the Cornus officinalis Sieb.et Zucc has obvious curative effect and the content of loganin and morroniside is high and stability,which can be the index compound,in this part,the method to test the content of loganin and morroniside by HPLC was established,which according to the 2015《Pharmacopoeia of the people,s republic of China》.The extraction times amount,times and time were determined by orthogonal test which base on the extraction rate of loganin and morroniside and paste-forming rate;the paste-forming rate is 36.24%,it is too high to make the granules,so we compared the centrifugation and alcohol sedimentation to the effect on the extraction rate of index component.The method and degree of concentrate and drying process conditions were determined by single factor investigation.The process of making Musu Granules was determined by study of molding process.2.Preliminary study on Quality Standard of Musu GranulesIn this part,Quality Standard of Musu Granules was study initially,which according to the request of the granules in 2015《Pharmacopoeia of the people’s republic of China》.Apart from ostreae concha and Cornus officinalis Sieb.et Zucc,the other nine herbs were identificated by TLC.The method to test the content of loganin and morroniside were established.Results1.Study on preparation of Musu GranulesThe preparation of Musu Granules is all herbs of Musu prescription are extracted two times,at the first time,they were extracted 60min by 10 times water and the extract is centrifugated 3000r/min for 30min.And the second time all herbs were extracted 60min by 8 times water and the extract is centrifugated 3000r/min for 30min.The second-time extract is added into the first-time extract which is concentrated,they will be taken into the vacuum drying oven when the relative density of the extract is 1.09～1.10g/mL,and they will be taken out after the content of water in the extract is under 5%.The Musu extract was grinding and passing through a No.4 sieve.The granules will be prepared with 90%ethanol and appropriate amount of dextrin.2.Preliminary study on Quality Standard of Musu GranulesIn this part,the qualitative identification method by TLC was established for five kinds of medicinal materials,which are Anemarrhena asphodeloides Bge、Cuscuta chinensis Lam、Lycium barbarum L、Ziziphus jujuba Mill.var.spinose(Bunge)Hu ex H.F.Chou and Ligustrum lucidum Ait.The method to test the content of loganin and morroniside were established.Take some Musu Granules,grinded and passed through a No.4 sieve.Took about 0.2g and added 10mL methanol,after 60min treatment with 50kHz and 300W ultrasonic treatment,passed through 0.45μm microporous membrane,the sample was prepared and used DIKMA Platisil C18(4.6mm×250mm,5μm),the condition of the determination was that acetonitrile for the mobile phase A,phosphoric acid for the mobile phase B with gradient elution.The elution conditions were as follows:0～20min 93%B,20～50min 93%→80%B;the speed was 1.0mL/min,the detection wavelength is 240nm,the temperature of the column was 35 ℃;the injection volume was 10μL。ConclusionsIn this study,preparation process and quality standards of Musu Granules were studied.A stable and feasible granules preparation process was established.Qualitative identification method by TLC was established for five kinds of medicinal materials,which are Anemarrhena asphodeloides Bge、Cuscuta chinensis Lam、Lycium barbarum L、Ziziphus jujuba Mill.var.spinose(Bunge)Hu ex H.F.Chou and Ligustrum lucidum Ait.The method of test the content of loganin and morroniside was established which was based on the《Chinese pharmacopoeia》.this study can provide basement of quality for controlling the quality of Musu Granules.Purpose and significanceCardiovascular disease is a series of diseases which is caused by heart and vascular diseases,it includes arrhythmia,hypertension,heart failure,viral myocarditis,coronary heart disease,angina and so on.With the socio-economic development and people’s lifestyles changing,the number of patients with cardiovascular diseases and the number of deaths which is coursed by cardiovascular diseases are increasing.At present,cardiovascular diseases have been one of the main reason of diseased deaths all over the world.The preparation of Compound Danshen is a famous medicine to treat the Cardiovascular disease,the effective of treatment is good.There are three herbs in this prescription,which are Salvia,Panax and Borneol,the water-soluble constituent has a poor absorption,such as the salvianolic acid B,the liposoluble constituen is hard to dissolute in vitro,such as tstanshinone;panax notoginseng saponins is unstability in acid,the mucosal permeability of Rbl and Rgl are very poor,these reasons effect the bioavailability of Compound Danshen preparation[3].Borneol has a good volatile,which can course the content of borneol reduce during the storage.Based on these problems,during the initial study,borneol β-CD inclusion was preparad to prevent the content reduce,and adhesion materials were selected and determined to prepare the Compound Danshen Adhesive Pellets,which had good spherical degree and adhesion ability,the surface area of the formulation is increased and residence time of the drug in the body is extended.Based on the previous studies,in this study,borneol and isoborneol would be the index compounds,and the dissolution in vitro and pharmacokinetics in vivo of the three kinds of Compound Danshen Pellets would be compared,at the last of this paper,the quality standard would be preliminary studied and the method to test the content and dissolution in vitro of borneol and isoborneol would be established,this study provide research basis for the Adhesive Pellets future development.Methods1.Determined the content of borneol and isoborneol of three kinds of Compound Danshen PelletsIn this part,the method to determined the content of borneol and isoborneol was established and compared the difference in content of borneol and isoborneol in the three pellets which had the same amount of feed.2.Compared the dissolution in vitro of the three kinds of Compound Danshen PelletsIn this part,the method to test the dissolution in vitro of borneol and isoborneol of the three kinds Compound Danshen Pellets was established which was based on the 2015《Pharmacopoeia of the people’s republic of China》and the dissolution curve was drawed.3.compared in vivo pharmacokinetics of the three kinds of Compound Danshen PelletsIn this part,the method to test the borneol and isoborneol in the plasma of rats by GC-FID was established.At first,10 times amount borneol(the normal administration amount is 0.6g/250g)was given to rats by gavage,we can determine the detection limit of sample and peak time of medicial concentration in blood.And then,0.6g/250g pellets were given by gavage and tested the borneol and isoborneol in the plasma of rats by GC-FID.The aim of this part was to compared the pharmacokinetics of three kinds of Compound Danshen Pellets.4.Study on content of borneol and isoborneol and the dissolution in vitro of BI-FDAP capsule.In this part,we used the pilot equipment to prepare the BI-FDAP and made the BI-FDAP to BI-FDAP capsule,because it is convenient to oral.the content of borneol and isoborneol was tested within 6 months,and the in vitro dissolution was tested in 0,6 months,we hope it can provide research basis on the quality standard of BI-FDAP capsule.Results1.Determined the content of borneol and isoborneol of three kinds of Compound Danshen PelletsIn this part,the method to test the content of borneol and isoborneol was established,the results showed that the linearity,precision,stability and repeatability were all meet the requirements.The sample recovery of BE-FDAP and FDP were meet the requirements,but the sample recovery of BI-FDAP was 95%below,it was coursed by the β-CD could adsorption the borneol.The raw material of three Pellets were the same,but the result of content test was different,the content of borneol and isoborneol in FDP was 6.15mg/g and 3.18mg/g,The content of borneol and isoborneol in BE-FDAP was 4.68mg/g and 2.54mg/g,The content of borneol and isoborneol in BI-FDAP was 6.52mg/g and 4.40mg/g,this result showed that the β-CD could prevent borneol to volatilize.2.Compared the dissolution in vitro of the three kinds of Compound Danshen PelletsIn this part,the result of the dissolution in vitro of borneol and isoborneol of the three kinds Compound Danshen Pellets showed the borneol and isoborneol in FDP and BE-FDAP can dissolute out completely in 40min in pH6.8 phosphate buffer,the cumulative dissolution rates of borneol were 103.78±7.40%、91.91±0.49%,the cumulative dissolution rates of isoborneol were 101.88±8.47%、100.96±2.29%.The borneol and isoborneol in BI-FDAP dissoluted out in 480min in pH6.8 phosphate buffer,the cumulative dissolution rates of borneol were 75.74±7.20%,the cumulative dissolution rates of isoborneol were 62.47±4.00%.the release speed of borneol in FDP and BE-FDAP were faster than the the release speed of borneol in BI-FDAP,and the cumulative dissolution rates of FDP and BE-FDAP is higher than the cumulative dissolution rates of BI-FDAP.It is showed the BI-FDAP has sustained release effect.3.Compared in vivo pharmacokinetics of the three kinds of Compound Danshen PelletsIn this part,10 times amount borneol(the normal administration amount is 0.6g/250g)was given to rats by gavage.The result showed that after administration 20min,the content of borneol and isoborneol in the the plasma of rats was 2.81μg/mL and 1.39μg/mL,then,0.6g/250g Pellets were given by gavage,the result of the test was that no detectable content was found for both borneol and isoborneol in the plasma,because the content of borneol and isoborneol was too low to be tested,and it was released slowly after the borneol was included by β-cyclodextrin.4.Study on content of borneol and isoborneol and the dissolution in vitro of BI-FDAP capsule.In this part,the content of borneol and isoborneol in BI-FDAP capsule during 2 months is stabilitable.The borneol and isoborneol in BI-FDAP capsule dissoluted out in 360min in pH6.8 phosphate buffer,the cumulative dissolution rates of borneol were 33.98 ±2.08%,the cumulative dissolution rates of isoborneol were 26.94±2.39%.The cumulative dissolution rates of BI-FDAP capsule is lower than BI-FDAP which was prepared in small experiment.Maybe,the adhesive material effected the dissolution of BI-FDAP capsule.ConclusionsIn first part of this study,a stable and good reproducibility method to test the content of borneol and isoborneol was established and compared the content of borneol and isoborneol of the three kind of Compound Danshen Pellets.Compared compared the difference in content of borneol and isoborneol in the three pellets which had the same amount of feed;in the second part,the method to test the dissolution in vitro of borneol and isoborneol of the three kinds Compound Danshen Pellets was established,the the dissolution curve was drawed;in the third part,the method to test the borneol and isoborneol in the plasma of rats by GC-FID was established.10 times amount borneol(the normal administration amount is 0.6g/250g)was given to rats by gavage,detection limit of sample and peak time of medicial concentration in blood were determined by this experiment,and then,0.6g/250g pellets were given by gavage and tested the borneol and isoborneol in the plasma of rats by GC-FID,but medicial concentration in blood was too low to be tested;in the fourth part,the content of borneol and isoborneol in BI-FDAP capsule was determined,the dissolution in vitro of borneol and isoborneol were tested after BI-FDAP capsule were prepared.The study of this research provided the basement to study the Compound Danshen Adhesive Preparations and the referece to study Modernization of traditional Chinese medicine preparation. |
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