| Objective:To build up the chemical fingerprints and compare the anti-ulcer effect of two species of ferula(Ferula ferulaeoides(Steud.)Korov.and Ferula assafoetida L.)Methods:1)Extraction rate of volatile oil and thermal stability of chemical compositions were selected as indicators,uniform design was used to optimize process of the steam distillation extraction of volatile oil from Ferula ferulaeoides(Steud.)Korov..The chemical components of volatile oil from two species of ferula were analyzed by GC-MS.2)The GC fingerprint for volatile oil from two species of ferula should be set up and be in line with the requirements of methodology.3)Yield rate and the total area of the main peaks were selected as indicators,orthogonal design was used to optimize the extraction process of water extracts from Ferula ferulaeoides(Steud.)Korov..The HPLC fingerprint of water extracts from two species of ferula should be set up and meet the requirements of methodology.4)Volatile oil and water extracts from two species of ferula were given orally with different doses to mice,acute toxicity symptoms and survival rate of experimental animals were observed for 14days to evaluate the acute toxicity of extracts from two species of ferula 5)In the experiments of ethanol-induced ulcer model,drug-induced ulcer model and acetic acid-induced ulcer model,a new image analysis method to calculate the inhibition rate of ulcer treatment should be established.And the image analysis processing method is verified by comparing with the classical method.6)According to the determination of inhibition rate(measured by classical method and image analysis method),the serum SOD activity,MDA content and the results of pathological test,anti-ulcer effects of extracts from two species of ferula were evaluated in the model of ethanol-induced ulcer,drug-induced ulcer and acetic acid-induced ulcer.7)Analgesic effects of extracts from two species of ferula were evaluated by writhing test.8)According to the bleeding time and coagulation time,hemostatic effects of extracts from two species of ferula were evlauted by cutting off mouse tail experiment and slide method.Results:1)The optimized extraction process of volatile oil from Ferula ferulaeoides(Steud.)Korov.were as follwing:adding 15 folds of water,extracting for 4h,the average extraction rate is 3.839%.30 kinds of compounds were identified in the volatile oil from Ferula ferulaeoides(Steud.)Korov.by GC-MS.The major components were guaiol(58.54%),farnesene(7.25%),nerolidol(6.08%),etc.25 kinds of compounds were identified in the volatile oil from Ferula assafoetida L.by GC-MS.The major components were carbonothioic dihydrazide(31.52%),fenadiazole(38.41%),(E)-3,7-Dimethyl-1,3,6-octatriene(8.32%),alpha-pinene(6.07%),etc.2)Two kinds of GC fingerprints for volatile oil from ferula were established in line with the requirements of methodology.Two kinds of GC fingerprints for volatile oil from ferula has 20 common peaks in which the fingerprint similarity was more than 0.9.3)The optimal extraction process of water extract from Ferula ferulaeoides(Steud.)Korov.was as follwing:extracting for 3 times,adding 6 times water every time,extracting 1.5h every time.Two kinds of HPLC fingerprints for water extract from ferula were established in line with the requirements of methodology.The HPLC fingerprint of water extract from Ferula ferulaeoides(Steud.)Korov.has 13 common peaks and that for Ferula assafoetida L.has 21 common peaks,the fingerprint similarity is more than 0.9.4)Volatile oil from Ferula assafoetida L.mainly lead to the acute poisoning symptoms of mice including the ankylosing convulsion and shortness of breath,the LD50 is 491.61mg/kg,95%confidence interval is 422.39-577.8mg/kg.Volatile oil from Ferula ferulaeoides(Steud.)Korov.mainly lead to acute poisoning symptoms of mice including the respiratory frequency decreases,the tail intermittent tilt,the LD50 is 10240mg/kg,95%confidence interval is 9140.4-11472mg/kg.Water extracts from Ferula assafoetida L.mainly lead to acute poisoning symptoms of mice including the twitching,shortness of breath.the LD50 is 17333mg/kg,95%confidence interval is 15536-19183mg/kg.The maximum tolerance of water extracts from Ferula ferulaeoides(Steud.)Korov.is 21000mg/kg.Compared with the extracts from Ferula assafoetida L.,the extracts from Ferula ferulaeoides(Steud.)Korov.have higher security.5)Compared with the traditional method,a similar result can be observed by the image analysis method,which is more objective and reliable,is useful to reduce the workload of test.6)Compared with the model group,the drug groups were able to make the test animal ulcer index and MDA content decreased,increase serum SOD activity(P<0.05).Volatile oil and water extracts from two species of ferula has certain inhibition on peptic ulcer according to different models tests.Anti-ulcer effect of volatile oil from Ferula ferulaeoides(Steud.)Korov.is more obvious.7)Compared with the blank control group,the writhing times of the mice in drug groups had significant difference(P<0.05),suggesting that two species of ferula have analgesic effect.The order of analgesic effect is as following:volatile oil of Ferula ferulaeoides(Steud.)Korov.>water extract of Ferula ferulaeoides(Steud.)Korov.>volatile oil of Ferula assafoetida L.>water extract of Ferula assafoetida L.8)Compared with the blank control group,there were no significant difference in the bleeding time and coagulation time of mice in drug groups(P>0.05),suggesting that the two species of ferula have no definite hemostatic effect.Conclusion:1)In this paper,the chemical fingerprint of volatile oil and water extracts from two species of ferula has been initially established,which has high precision,good repeatability and stability,and provides the experimental basis for the two ferulic medicinal materials quality control.2)Compared with the traditional measuring method,the image analysis method has more advantages for determination of damage evaluation.3)The extracts from two species of ferula have clear pharmacological activities,and Ferula ferulaeoides(Steud.)Korov.has the potential advantage of clinical application.Ferula ferulaeoides(Steud.)Korov.may be a substitute of Ferula assafoetida L.. |
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