| ObjectiveLiangyuetan is a resinous woody cane from Dalbergia hent.hamii Prain.It mainly produced in China’ s Hainan,Guangdong,Guangxi and other regions,Southeast Asia and other places are also distributed.It has the effects of promoting blood circulation and dredging the channels and collaterals,and can be used to treat bruises,pains in muscles and hones,irregular menstruat ion,and dysmenorrhea,etc.Hainan Li people and other folks used Liangyuetan to maketlea or soak in wine.Orally,it was used to prevent and treat cardiovascular and gastrointestinal diseases.Topical application was used as a hemostatic,anti-inflammatory,and myogenic drug for wounds such as kni fe wounds,burns,and snake bites.Inaddition,due to its aromatic smell,Liangyuetan is commonly used to incense,remove mosquitoes,purify the air,and fight against plague.It is often used to make precious spices and crafts in the fragrance industry in China.Modern research shows that Liangyuetan has various pharmacologicogical activities such as antibacterial,anti-inflammatory and antioxidant.As a commonly used traditional Chinese medicine in southern China,Liangyuetan has a long history of dual-use medicine.However,there is currently little research on Liangyuetan,and the Chinese Pharmacopoeia and local standards are not included.Therefore,in order to reasonably develop and utilize the medicinal resources and ensure the safety of clinical medicine,this subject takes Liangyuetan as the research object,adopts tradi tional and modern scientific and technological means to examine the original plant morphology,t.raits,microscopic and thin layer chromatography items and extracts for systematic research.This article also explores its volatile oil and total flavonoid content,volatile oil chemical composition,total flavonoid extraction process and antibacterial effect of volatileoil.Finally,,draft quality standards for Liangyuetan were prepared,and quality standards for Liangyuetan were init ially established.Methods1.Study on pharmacognosy identificationThe morphological characteristics of the original plants of Liangyuetan were studied by the original plant identification method.The characterist ics of Liangyuetan were summarized by the traditional identification methods such as eye view,hand touch,nose smell and mouth taste.The microstructure,powder and dissociated tissue were studied by the microscopic ideatification method.A TLC method for the identification of Liangyuetan was established by investigating the extraction and expansi on condition.2.Inspection items and extractionAccording to the method specified in the general principles of the fourth part of Chinese Pharmacopoeia(2015 Edilion),the content of water,total ash,water-soluble and alcohol-soluble ext racts of 30 batches of Liangyuetan from different habitats were determined.According to the test results,the content limi ts of water,total ash and extract of Liangyuetan were established.3.Determination of contentThe volatile oil of Liangyuetan from different areas was extracted by steam distil lat ion and its content was determined.With rutin as the control,the total flavone content of Liangyuetan from different areas was determined by UV spectrophotometry.4.GC-MS analysis of volatile oilsGC-MS method was used to analyze the chemical consti tuents of the essential oil in Liangyuetan from different habitats.5.Study on the extraction process of total flavonoids from LiangyuetanOn the basis of single factor test results,response surface method was used to optimize the extraction process of total flavonoids from Liangyuetan.6.Antibacterial effect of volaitile oil from LiangyuetanFilter paper diffusion method was used to screen the antibacterial activity of the essential oil of Liangyuetan,and the minimum inhibitory concentration of volatile oil from Liangyuetan to Candida albicans was determined by double di lution method.Results1.Study on pharmacognosy identificationThe main identification characteristics of the plant morphology,trait microscopy and TLC of Liangyuetan were summarized.(1)The distinguishing features of the original plants are as follows:pinnate compound leaves,2-3 pairs of leaflets,nearly leathery,ovate or elliptic,blunt and emarginate apex,wedge-shaped base,glabrous above and slightly puberulent below;inflorescences axillary,calyx campanulate,corol la white;pods thinly leathery;kidney shaped seeds.(2)The characteristics of trait identification are irregular cyl indrical or long pieces.The surface wood grain is thick and obvious,showing light brown,red brown or brown black.Small holes in the duct can be seen on the section,with floral,honey and coconut fragrance various special aromas.(3)The microscopic identification features are as follows:1-4 rows of wood ray cells in the transverse section,2-3 rows more,scattered vessels,multiple single holes,occasionally 2-11 double tube holes in the radial section;non overlapping wood ray cells in the tangential longitudinal section,1-4 rows in the width,2-3 rows more,5 sparse rows,and 5-35 cells in the height.Wood ray cells in the radial longi tudinal section are heterogeneous,arranged horizontally in strips,and overlapped in the axial parenchyma tissue.Wood fiber,vessel,ray,parenchyma cell and calcium oxalate square crystal can be seen in the powder.In the dissociated tissue,there were wood fiber cells,vessel cells,wood parenchyma cells and wood ray cells.(4)A TLC system for the determinination of Liangyuetan was established.Toluene:ethyl acetate:acetic acid(9:3:0.02)was used as the developing system,Liangyuetan reference materials as the control,10%sulfuric acid ethanol solution as the color developing agent.The results showed that after the sample was developed,the separation was good and the spots were clear.2.Inspection items and extractionIn the 30 hatches of the Liangyuetan samples,the moisture determination range was 7.03%to 12.98%,with an average value of 9.30%.The total ash content ranges from 1.26%to 6.33%,with an average value of 3.29%.The range of water-soluble extractives is from 2.80%to 7.58%,the average value is 5.10%.The content of alcohol-soluble extractives ranged from 28.50%to 48.56%,with an average of 39.39%.According to the measurement results and the actual production situation,the inspection items and the limils of extracts of Liangyuetan were drawn up.Moi sture shall not exceed 13.0%,total ash shall not exceed 7.0%,water-soluble extract shall not be loss than 3.0%,and alchol-soluble extract shall not be less than 28.0%.3.Determination of contentThe content of volatile oil in 30 batches of Liangyuetan medicinal materials is from 0.28%to 2.37%.Based on the measurement results,theyield of the volatile oil of Liangyuetan is preliminari ly formulat ed to be not less than 0.30%(mL/g).The total flavonoids content is 1.18%～6.25%.4.GC-MS analysis of volatile oilsA total of 109 compounds were identified in the 30 batches of volatile oil from Liangyuetan medicinal materials.The oil mainly contained sesquiterpenes,alkenes,alcohols,ethers,esters and other compounds.The main ingredient is elemicin,followed by Methyleugenol,and the sum of the two contents accounts for more than 60%of the total ingredient content.In addition,it still contains trace components such as 1,2-dimethoxy-4-N-propeny lbenzene,linalool,whiskey lactone,isolimarin,cis-β-cubene and other trace ingredients.5.Study on the extraction process of total flavonoids from LiangyuetanThe best extraction conditions of total flavonoids from Liangyuetan were temperature 50 ℃,extraction time 60 min,ethanol vol ume fraction 60%,liquid to material ratio 50:1.The extraction rate of total flavonoids from Liangyuetan was 4.51%.6.Antibacterial effect of volatile oil from LiangyuetanThe volatile oil of Liangyuetan has bacteriostatic effect on Staphylococcus aureus,methicillin-resistant Staphylococcus aureus and Candida albicans in different degrees.Its minimum bacteriostat ic concentration against Candida albicans is more than 12.50 mg/mL.ConclusionThis topic systomatically researched the quali ty standards of Liangyuetan,and drafted the draft quality standards of Liangyuetan.The plant morphology,trai ts and microscopic identification characteristics of Liangyuetan plants established are accurate and obvious.The identification of thin layer chromatography is simple,the separation effect is good,and the characteristic is strong.The proposed moisture,total ash and leach content Iimits are reasonable.The determination method of volatile oil and total flavonoids is easy to operate,the results are accurate and the reproducibility is good.The optimal process of total flavonoid extraction is stable and feasible,and the operat ion is simple.Therefore,the research results of this paper can provide a theoretical basis for quality control and in-depth research of Liangyuetan.At the same time,it can provide a reference for the rational development and utilization of private medical resources. |
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