Experimental Studies On The Quality Control Of Miao Medicine-Prionobelum Qianensis Chen

Prionobelum qianensis Chen,is the dried whole insect of Zephroniidae,Prionobelum.Most of them perched in moist places on hillsides,under dead branches and saproot leaves or stones,which was mainly in Guizhou,Yunnan and Sichuan provinces.P.qianensis is medicinal whole insect,with the effect of clearing away heat and detoxifying,dispelling wind and clearing collaterals,dispersing stasis and relieving pain,mainly used to treat rheumatism and arthralgia,vein clonus,injury and swelling of fracture and so on.It is recorded in Chinses Herbal Medicine-Miao,Guizhou Miao nationality Medicine Research and Development,etc.Its clinical application about inflammation and tumor is extensive and the curative effecs is very remarkable in the folk,especially in Guizhou Miao nationality areas.However,the research on the quality standard,chemical composition,pharmacodynamics and substance basis of P.qianensis is relatively weak both here and abroad and the lack of systematic quality control standard system have seriously restricted the scientific system development and extensive use of P.qianensis.In order to make up for the above problems,the volatile components,main amino acid composition and content determination,and quality standard of P.qianensis were studied in this paper.The main results are as follows:1.The analysis of volatile components and health risk assessment of P.qianensisIn this part,the volatile components were analyzed by GC-MS.The results showed that the volatile components mainly contained alcohols,aldehydes,pyrazines,sulfides,benzothiazoles,amines and pyrrole compounds.Their contents were 64.09%,15.61%,10.47%,4.45%,3.78%,1.50%and 0.10%。The principal component analysis and cluster analysis of the main volatile components of P.qianensis,the results showed that they were classified into 7 principal components.The characteristic values were 3.987,3.580,3.232,3.188,3.142,2.267 and 1.433;The cluster analysis showed that the 10 batches of P.qianensis were mainly divided into two groups:S₁（Huaxi,Baihuahu,Wengan,Anshun,Qingyan,Longli）and S2（Bijie-qianxi,Tongzi,Hongfenghu,Bijie-hezhang）.The grey correlation analysis between the volatile components of P.qianensis and the environmental factors,including the average temperature,altitude,the content of precipitation,O₃,NO₂,SO₂,CO and inhalable particles of the collecting sites of P.qianensis.The results showed that environmental factors had an important effect on the species and content of volatile components of P.qianensis.The safety of volatile compounds of P.qianensis has been preliminarily confirmed through the data of the National Food Safety Standard-Food Additive Standard（FEMA-GRAS）and the authoritative organizations such as International Organization of the Flavor Industry IOFI-Global Reference List of Chemically Defined Substances.The health risk evaluation of benzaldehyde,the most important volatile component in the volatile components of P.qianensis,was carried out by the health risk assessment model of the US Environmental Protection Agency.The results showed that the risk entropy of benzaldehyde was 1.646×10^-6 mg·kg^-1·d^-1,which was much lower than the limit of benzaldehyde0.100 mg·kg^-1·d^-1.We can confirm the safety of volatile components to the human body.2.Determination and Analysis of 15 kinds of Amino acids in P.qianensisIn this part,the Venusil AA amino acid column was used to determine the content of 15kinds of amino acids in P.qianensis.The optimum hydrolysis and derivatization conditions were determined by single factor and orthogonal test:hydrolysis time 80 min,derivative reagent dosage 3 mL,derivatization temperature 30℃,and derivative time is 30 min.A method for the determination of 15 kinds of amino acids in P.qianensis was established and the method was used to determine the amino acids in 11 different places in Guizhou Province.The total amino acid content of the total amino acids varied from 114.81 mg·g^-1 to145.11 mg·g^-1.The contents of amino acids in 11 batches of P.qianensis were analyzed by principal component analysis and cluster analysis.The results showed that the significant or extremely significant correlations between the other amino acids except tyrosine,methionine and cystine.Principal component analysis showed that three characteristic values greater than 1,were 9.552,2.170 and 1.194.Their cumulative variance contribution rates were 63.680%,78.147%and 86.107%,respectively.The results of cluster analysis showed that 11 batches of P.qianensis were divided into two categories:S1:Hongfenghu,Qianxi,Baihuahu,Anshun,Qingyan,Hezhang,Huaxi and Tongzi and S₂:Longli,Huishui and Maige.3.Experimental studies on the quality control of P.qianensisAccording to the general rules of Chinese Pharmacopoeia 2015 edition,the methods of quality standard research were used to study the water content,ash content,extract,heavy metal content and pesticide residue of P.qianensis;using phenylalanine and alanine as the reference substances,the TLC identification of P.qianensis showed that the color spots were clear,no tail and the degree of separation was good.To establish a HPLC method for the determination of glycine,alanine and proline,which can be used for the quality control of P.qianensis,and to investigate its methodology and to draw up a draft quality standard for medicinal materials,string to include it in the new edition of“Guizhou Province traditional Chinese medicinal quality standards”and lying a solid foundation for the scientific development and extensive utilization of the materials of P.qianensis Chen.