| Chuankezhi Injection is traditional Chinese medicine injection which composed of Morinda officinalis and Epimedium davidii.The raw material of the injection is Morinda offcinalis oligosaccharides(MOO)from M.offcinalis and flavonoids from Epimedium Extract.The injection have effects of relieving asthma and cough,antianaphylaxis,improving the function of hormones immunity and cell-mediated immune competence in pharmacological research.It uses clinically in the symptom of asthma and capillary bronchitis.The quality standard is recorded in the New Positive Standard of the National Drug Standards(volume 65).The standard includes qualitative and quantitative identification of E.davidii,but lack of the investigation of medicinal materials of M.offcinalis and MOO in the Chuankezhi Injection.For this purpose,this paper is going to evaluate the quality of original injection and its production process in the present,and find every factor affect the MOO extract quality in Chuankezhi.It carried out studies of MOO extract method of M.offcinalis,chemical stability,and concentration or dry temperature in the production process.Moreover,this research had established the production process of MOO extract in Chuankezhi injection.On this basis,we has built the standard criterion of the MOO in M.offcinalis(bulk drug),MOO(intermediate products),and Chuankezhi Injection(finished product).Details were as follows.Part one Documentary ResearchThis part had consulted related documents on M.offcinalis,Chuankezhi Injection and Chinese medicine injection fingerprint in order to illustrate the significance of using MOO as specification.Part two ExperimentChuankezhi Injection Quality AssessmentUsing high performance thin layer chromatography(HPTLC)and high performance liquid chromatography-evaporation laser scatter detector(HPLC-ELSD)to assess the quality of Chuankezhi injection and MOO extraction production technics.Results were as follows.1.Comparing MOO extract with MOO from bulk drug,the concentration of Sucrose and 1-kestose were higher than MOO from bulk drug obviously,while Nystose and 1F-Fructofuranosylnystose were lower than MOO from bulk drug.2.The MOO from bulk drug continually declined during extraction,concentration,and desiccation.It may be the reason for low content of MOO in Chuankezhi Injection.In order to investigate the factor that affects the quality of MOO,studies of extraction method,chemical stability,concentration or dry temperature to bulk drug must be carry out systematically.Using HPTLC to study the stability of MOO from M.offcinalis during extractionIn this paper,HPTLC was used to investigate the stability of MOO from M.offcinalis extracted in water,ethanol,and different pH of water.The results were as follows.1.The MOO from bulk drug was not stable when extracted in water.So it is suggested that it should not be extracted in water.2.The extraction ratio reached the highest when the percentage of ethanol was in the range of 50%to 60%.3.When the bulk drug extracted in different pH of water,it would be completely or partly hydrolyzed in the range of pH between 2 with 5 and stable when pH was above 6.So it is suggested that it should be extracted in neutral environment.Using HPTLC to study the stability of MOO from M.offcinalis during extractionIn this paper,HPTLC was used to investigate the stability of MOO from M.offcinalis extracted in water,ethanol,and different pH of water.The results were as follows.1.When the bulk drug extracted in water,the higher degree of polymerization(DP)of MOO,the easier to be hydrolyzed.It is suggested that M.offcinalis do not extract in water.2.When the bulk drug extracted in ethanol,the extraction ratio reached the highest when the percentage of ethanol was 50%.It is suggested that M.offcinalis should be extracted in 50%ethanol.3.MOO was strongly influenced by pH during extraction.When the pH is 2 to 4 in water,it would be completely or partly hydrolyzed.The chromatography started to stable when the pH above 8.The reason for it may be the low pH or phosphate buffer make the baseline unsmooth.4.Ethanol had protective effect to MOO when it extract in 50%of ethanol.It is recommended that MOO is better extracted in 50%of ethanol.Study of MOO production procedure from bulk drug of M.offcinalisI The extraction procedure of MOO from bulk drug of M.offcinalisAccording to the research results,establishing the production procedure from bulk drug of M.offcinalis,and got MOO total extracting solution.In order to study the influence of temperature in concentration of MOO extracting solution,1L solution was took to concentrate to dr.y in 50?,60?,70?,80?,90? through rotary evaporators in 0.1MPa.Using HPLC-ELSD assay to the content of Sucrose,1-kestose,Nystose,and 1F-Fructofuranosylnystose.Result showed that MOO stable at the temperature range from 50? to 90?when the pH was 6 to 7 during concentration.II The pure procedure of MOO crude extractMOO plaster had made from the established pure procedure.In order to study the influence of temperature in desiccation of MOO crude extract,0.15 gram MOO crude extract was took to dry at the temperature of 60?,70?,80?,90?,100?,110? in oven.Using HPLC-ELSD assay to the content of Sucrose,1-kestose,Nystose,and 1F-Fructofuranosylnystose.Result showed that MOO didn,t have significant change(p>0.05)except Sucrose and 1-kestose.While the dry temperature is above 100?,MOO changed to yellow.So the desiccation temperature is 60? to 80?.The stability test of MOOMOO stability test was carried out according to the stability testing of pharmaceutical preparations and guiding principles.MOO 0.15g was placed in high temperature(6O?),high humidity(RH 75%±5%)and strong light(4500± 500LX)environment 30 days.Sampling carried out in 1st,2nd,3rd,5th,7th,10th,20th,30th day,HPLC-ELSD to assayed content of Sucrose,1-kestose,Nystose,and 1F-Fructofuranosylnystose.The result showed that MOO would have change in the environment described above.1.MOO would change yellow since 7th day when stored in high temperature,and the content of 1-kestose,Nystose,and 1F-Fructofuranosylnystose would declined since the 2nd day.2.MOO would be hygroscopic and gain weight in high humidity,and Nystose,and 1F-Fructofuranosylnystose would descended.3.MOO would decline when stored in strong light.Quality standard establishment of Chuankezhi InjectionI It had established 10 bath of MOO fingerprint in bulk drug of M.offcinalis(4 to 5 year).The similarity analysis indicated the similarity between 10 bathes of bulk drug.The assay of different growing years of M.offcinalis showed that the content of Nystose(3%to 6%)in 1 to 6 years-grow bulk drugis higher than the pharmacopoeia criterion(2%).However,the total MOO is higher in 4 to 5 year-grow bulk drugs.? The fingerprint of MOO extract had few impurity,which means the purity of the MOO extract.? It had established 10 bath of Chuankezhi Injection fingerprint.The similarity analysis indicated the stability between bathes.Comparing the Chuankezhi Injection with MOO,the oligosaccharides in DP had omitted,which lead to 13 characteristic peaks indentified.? Nystose and 1F-Fructofuranosylnystose were higher in Chuankezhi Injection than original Injection,and the kind of MOO were higher,too.Conclusion? The content of the MOO in original Chuankezhi Injection was low,may be related to the extraction,concentration,and purification process in production procedure.? The stability study of MOO during extracting showed that bulk drug of III M.offcinalis should be extracted in 50%of ethanol.The improved production procedure of MOO extract was stable,and the yield of MOO was 31%.? MOO extract is better to put in cold,dry and dark environment.Standard criterion of the MOO in M.offcinalis(bulk drug),MOO(intermediate products),and Chuankezhi Injection(finished product)had been established. |
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