| Objective:Hantaoye also called Qiyelian,Qijiapi,Qiyelan, Long-shoushu,and so on. It is belonged to Araliaceae section Schefflera kwangsiensis Merr.ex Li category plant,located in Guangxi,Yunnan, Guizhou, Fujian, Zhejiang Province of China. It is a traditional herbal medicine in the Ethnic minority areas such as Miao nationality and Li nationality, It is mainly used to relieve pain, such as trugeminal neuralgia, sciatica and gastrointestinal colic,especially to some neuralgia, and with the characteristics of little side effects.According to the civil application experience and preliminary reported in the literature,e activity constituent of analgesia and anti-inflammatory in Hantaoye is organic acid. The research object is Hantaoye(chefflera kwangsiensis Merr.ex Li).The purpose of research is to establish the quality standards of Hantaoye, and to study the optimum extraction of organic acid from Hantaoye. And apply to modern separation and identification techniques to study chemical composition of organic acid. Develop the crude ointment of Hantaoye and conduct a study of the analgesia and anti-inflammatory medicinal experiment to provide pharmacological experiments basis for the further development of the new formulation and reasonable security clinical medicine.Method:1. Using HPLC, fumaric acid as the standard reference substance,to establish the quality standards, to clear the method for determination of organic acid and using orthogonal method to design the optimum extraction of organic acid from Hantaoye.2. Combined with Ion exchange chromatography and HPLC to extract organic acids from Hantaoye, and using modern separation and identification techniques to separate and identify the chemical compositi-on of organic acid.3. Based on the physical and chemical properties of organic acid, produce small scale W/O types ointments of extraction and organic acid. Establish the analgesia and anti-inflammatory medicinal experiment methods:(l) The hot tingling test and acetic acid writhing method were adopted in the experiment to study the analgestic effect of Hantaoye ointments;(2) xylene-induced ear swelling, formaldehyde-induced hind paw edema and the level of PGE2in the paw was examined were prepared to study the effects of Hantaoye ointments, to evaluate its mechanism.Result:After the inspection of HPLC, the results showed tahat the method is accurate, stable, good reproducibility,precision and good recovery.Using the orthogonal method, fumaric acid content as the index, explored the best extraction process is60%ethanol concentration, extraction time of3h,2times and10times of the liquid ratio. The yield of organic acid in Hantaoye is2.655%by the Ion exchange chromatography. Derived by melting point determination and HPLC to identify the two compounds were separated from the organic acid:compound I is Mucic Acid,ompound II is Fumaric Acid.After the7d of continuous topical smear administration by Hantaoye groups ointments, the results showed the analgestic effect. The hot tingling test showed that the high extraction group and the high acid group at60min after administration, compared with model group, there was a most significant difference(P<0.01), the middle acid group has a significant difference(P<0.05).At90min after administration, the high acid group, compared with model group, has a significant difference(P<0.05). In the mouse writhing test, the high acid group has a most significant difference(P<0.01), the high extraction group, compared with model group, there was a significant difference(P<05).The xylene-induced ear swelling results showed that the high acid group has a most significant difference(P<0.01), the high extraction group and the middle acid group, compared with model group, there was a significant difference(P<0.05). In the formaldehyde-induced hind paw edema, the high acid group and the middle acid group has a most significant differenceP<0.01), the high extraction group there was a significant difference(P<0.05), compared with model group. From the level of PGE2in the paw showed that the high acid group has a most significant difference(P<0.01), the high extraction group,the middle extraction group and the middle acid group, compared with model group, there was a significant difference(p〉0.05). It showed that Hantaoye has the effect of anti-inflammatory.Conclusion:The study of quality standard of Hantaoye is accurate and effective. It has the effects of analgesic and anti-inflammatory, the active part is organic acid. The anti-inflammatory mechanism maybe related to the inhibition of PGE2production, enhance immune function. The new formulation of Hantaoye ointments is worth further research,development and utilization. |
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