Study On Quality Standard Of The Stem Of Melicope Ptelefolia

This paper is divided into six parts,including the quantity analysis and the quality study of the stem of Melicope ptelefolia,also,including the preliminary draft on the quality standards.ObjectiveThe Melicope ptelefolia,at present,is still broadly used in many of the Chinese patent medicine,for instance,the“san-jiu-wei-tai,san-jiu-gan-mao-ling”,the others have using the plant for a long time.But the differences of the places of origin source and the packing time maybe influence the clinical effect deeply.For the time being,there are only a few studies reporting on the quality of the Melicope ptelefolia at home and abroad,and maybe including the management mechanism in the pharmaceutical market is not consolidated.Therefore,it is quite necessary to study the quality so that the departments of quality assurance and manufacture factories can have an effective quality control method.Based on the present research on the chemical component,we need to establish the quality study of the Melicope ptelefolia to perfect the research and hope to darft the quality standards on the stem of Melicope ptelefolia.Method1.The study on the identificationThe method of microscopical identification was used to analyze the characteristics of the powder of the Melicope ptelefolia stem,and the TLC also was used as the control method for the qualitative identification with the alkaloids as the reference substance2.The quality inspection of moisture?ash content and extractumUsingthe drying method in the general principles of 0832 recorded in the 2015 China pharmacopoeia to measure the content of moisture about 16 samples;using the method in the general principles of 2302 to test the content of ash;the method recorded in the general principles of 2201 is used to measure the content of extractum.The results can be supported to establish an effective quality control method.3.The research on the chemical composition contentsTo establish a quantitative method,optimizing the chromatographic conditions and conducting the methodology,we use the method of HPLC to measure the contents of evolitrine?dictamnine?kokusaginine and 4-methoxy-2?1H?-quinolinone in the 16 batches of the stem of Melicope ptelefolia.4.Study on the contents of total alkaloidsTwo methods are used to study the contents of total alkaloids of 16 batches,the back-titration and the UV-spectrophotometry which have been recorded in the 2015 China pharmacopoeia.And the methodology was also examined and compared.The results can be used to estimate the optimal harvest time and the place of the Melicope ptelefolia.ResultsThis experiment was studied comprehensively by the qualitative identification and the quantitative analysis to evaluate the quality of the stem of the Melicope ptelefolia,all the experiments could provide us the results to analyze the data to draft the quality standards premilinarily.It was found that the method of microscopical identification for powder and the method of TLC for the qualitative identify,the specificity was enhanced compared with the single reference substance as contrast method.We used the drying method in the general principles of 0832 recorded by the 2015 China pharmacopoeia to measure the content of moisture,it couldn't be more than 10%.The method was used which has been recorded by the general principles of 2302 to to test the content of ash,it suggested that the content of ash couldn't go beyond the 5%.Both water-soluble extraction and alcohol-soluble extraction were tested with the method of the general principles of 2201.The determination of water-soluble extraction and alcohol-soluble extraction used by hot-soak method couldn't be less than 3%,the contents used by cold-soak method should higher than2%.The HPLC method was performed on the Kromasil C₁₈8 column?250mm×4.6mm,5?m?with the column temperature 35?,at the flow rate of 1.0 mL·min^-1.The absorption wavelength was 210nm.We used the methanol-water as the liquid phase and the gradient elution was performed and the sample size was 10?L.We had tested the contents of four compounds of alkaloids,including the evolitrine?dictamnine?kokusaginine and the 4-methoxy-2?1H?-quinolinone.The gradient elution time was 120 minutes.In order that we can make a comparative analysis,we just considered the samples of S8-S16.The content of evolitrine is between 0.146‰and 0.282‰,the range of the content is a relatively large change.The content of the dictamnine is 0.095‰to 0.211‰,the variation range of the content is obvious.However,the content of kokusaginine in the16 samples of the stem of Melicope ptelefolia is the lowest,between 0.083‰and 0.122‰.The fourth reference substance,4-methoxy-2?1H?-quinolinone,the content is relatively stable,the range of the content is between 0.120‰and 0.169‰.It turned out that there was a large gap in the 16batches of the stem of Melicope ptelefolia,but the results of samples 8 to 16 were similar.Two methods were used to measure the content of total alkaloids of the 16samples,the back-titration and UV-spectrophotometry,and the methodology was alsoexamined.According to the experiment data,and making sure the stability of sample source,we suggest that the optimal harvest time and place is the fourth quarter of Qing-yuan,and the better method to study the contents of total alkaloids is titration.According to the requirements of the 2015 China pharmacopoeia,combining with the results,we could draft the quality standards of the stem of Melicope ptelefolia.ConclusionThe basic quality standard researches of the stem of the Melicope ptelefolia were established comprehensively.The method is accurate and sample,the repeatability and the stability are good,which the mentioned above could provide an experiment parameters for the study on the stem of the Melicope ptelefolia.