| Object:Based on the theory of traditional Chinese medicine,thepharmacological identification method was used to compare the·traits of thetangerine peel contained in the Pharmacopoeia,and the thin layer identification method of four tangerine peels was established by thin layer identification.The method for determining the content of the effective components of tangerine peel and the combination of pharmacological effects were established.Experiments,as well as the comparison of four kinds of dried tangerine peel oil by GC-MS and the combination of pharmacological experiments,the comparative study of four kinds of tangerine peels in the pharmacopoeia,in order to provide reference for the search for effective substitutes of dried tangerine peel and to develop clinical application and quality standards of tangerine peel.Provide scientific evidence.Method:1.Using the method of biopharmaceutical identification,the microscopic identification and trait identification of the four varieties of dried tangerine peel contained in the Chinese Pharmacopoeia were carried out,and the traits and microscopic differences of the tangerine peels of the four varieties were compared.2.Through the thin layer identification methods of citrus genus such as tangerine peel,clam shell and orange-red,contained in the Pharmacopoeia,the above three medicinal materials were thinly identified,and the thin layer of three Chinese medicinal materials was carried out by polyamide film chromatography.Check and compare the two methods.3.Establish HPLC method for determination of the main active components in dried tangerine peel,and combine the normal small intestine propulsion and cough expectorant experiments to compare and compare the differences of the four tangerine peels contained in the pharmacopoeia.4.Using modern extraction analysis technology,extract the volatile oil from the four tangerine peels contained in the Pharmacopoeia,and analyze the differences of the components in the volatile oil between different tangerine peels by GC-MS analysis.5.Taking the volatile oil from the four dried tangerine peels contained in the Pharmacopoeia as the test sample,the effect on the isolated small intestinal smooth muscle of rabbits was to find the active ingredients consistent with the indications of the tangerine peel function,and to explore the dose-response curve of rabbit isolated small intestinal smooth muscle.Relationship,and preliminary discussion of its pharmacodynamic mechanism.Restult:1.In the aspect of trait identification,it can be distinguished by the thickness of the peel,the difference of the odor and the size and intensity of the oil chamber of the fluoroscopy,while other features also play a certain auxiliary role in the identification of the tangerine peel varieties;In terms,it can be distinguished by the morphology and quantity of hesperidin crystals and calcium oxalate crystals.2,according to the 2015 edition of the"People's Republic of China Pharmacopoeia"Part IV:thin-layer chromatography?General Rule 0502?test,the thin layer method contained in the Pharmacopoeia can not be the hesperidin,neohesperidin and naringin in the three herbs Completely distinguished,polyamide film chromatography can simultaneously identify hesperidin,neohesperidin and naringin in three Chinese herbal medicines.3.The method for determining the content of the main active ingredients of dried tangerine peel was established,and the content of hesperidin,nobiletin,heptamethoxyflavonoid and hesperidin in the four varieties of dried tangerine peel contained in the pharmacopoeia was determined.Combined with normal mouse small intestine propulsion and cough sputum test,Dahongpao,Chazhigan,Wenzhou mandarin and Futang decoction have promoted the small intestine propulsion in mice,and all have certain cough and expectorant effects.Among them,the content of the above four components in the tea citrus and Dahongpao is similar,but the promoting effect of the small intestine of the tea citrus is the most significant and the cough sputum effect is the best.According to the experimental results of the extraction of volatile oil from tangerine peel,the yield of volatile oil from Dahongpao was 3.47%,the yield of volatile oil from T.chinensis was 4.54%,and the yield of volatile oil from T.citrifolia was 3.57%.6.14%;The total ion chromatogram of volatile oil from Dahongpao tangerine peel samples from Jiangxi Xingan was obtained by GC-MS.It was found that the four tangerine peels contained in the Pharmacopoeia contained terpene,citronellal,terpinolene,geranyl,and?.-decene,?-terpinene,b-elemene,?-pinene,?-myrcene,D-limonene,g-terpinene,g-elemene,?Z?-caryophyllen,etc.13 A common ingredient.By using the past software to perform principal component analysis and cluster analysis on GC-MS data,the tangerine peel can be roughly divided into two types:Guangdong Xinhui tea citrus samples and other regional samples.5.Pharmacological test Different concentrations of volatile oil have certain relaxation effects on different parts of isolated small intestinal smooth muscle.The final concentration of1×10-5 mL/mL volatile oil of dried tangerine peel has the most obvious effect on the relaxation of isolated small intestine.The maximum inhibitory concentration of the isolated small intestine;compared with the other two types of dried tangerine peel,the inhibitory effect of Fudan and Citrus essential oil on the duodenum was significant,and it had a dose-effect relationship?*P<0.05,**P<0.01?.Both acetylcholine and histamine phosphate can induce isolated intestinal muscle tendon,and the maximum concentration of acetylcholine on isolated intestinal muscle of rabbit is 5.0×10-7 mol/L?ie,the concentration of 1.0×10-3 mol/L is administered?.,10?L),the maximum cariogenic concentration of histamine phosphate on rabbit intestinal muscle is1.5×10-5 moL/L?ie 1.5×10-2 moL/L,20?L?;the volatile oil of tangerine peel can effectively inhibit acetylcholine or histamine phosphate Caused by small bowel fistula.Conclusion:1.The traits and microscopic characteristics of the four tangerine peels contained in the Pharmacopoeia can be compared by trait identification and microscopic identification methods,and the four tangerine peels contained in the Pharmacopoeia are initially identified.2,according to the 2015 edition of the"People's Republic of China Pharmacopoeia"Part IV:thin-layer chromatography?General Rule 0502?test,the thin layer method contained in the Pharmacopoeia can not be the hesperidin,neohesperidin and naringin in the three herbs Completely distinguished,polyamide membrane chromatography can simultaneously identify hesperidin,neohesperidin and naringin in three herbs.Using chloroform-methanol-acetone=15:3.5:1.5 as developing solvent and 10%aluminum trichloride in ethanol as the color developing agent,hesperidin and neohesperidin in dried tangerine peel,clam shell and orange-red extract The flavonoid glycosides such as naringin have good qualitative and qualitative identification on polyamide membranes,and the separation effect is good and the spots are round.The method is simple,rapid,clear,accurate and reliable,with strong specificity and good versatility.It can be used as a qualitative identification method for hesperidin,neohesperidin and naringin in tangerine peel,clam shell and orange.3.By establishing a method for determining the content of the active ingredients of dried tangerine peel,combined with pharmacological experiments,a comparative study was conducted on the four tangerine peels contained in the pharmacopoeia to find other varieties of tangerine peel that are similar to the dried tangerine peel.4.By using the past software to perform principal component analysis and cluster analysis on GC-MS data,the dried tangerine peel can be divided into two types:Guangdong Xinhui tea citrus fruit samples and other regional samples,which can be used for Xinhui tangerine peel and other varieties of tangerine peel.Identification.5.The volatile oil of dried tangerine peel can inhibit the intestines caused by histamine or acetylcholine.It indicates that the inhibition of gastrointestinal tract by the volatile oil of tangerine peel may inhibit the gastrointestinal motility or directly by affecting the secretion of histamine and acetylcholine.Acts on gastrointestinal smooth muscle to reduce the frequency of gastrointestinal motility. |
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