Study On The Scientific Connotation Of "Euodia Rutaecarpa,the Older The Better"

Objective: Euodia rutaecarpa(ER)is one of the "Liuchen",and it is known as "the older,the better".However,there is few reason for it.In the early stage,we have systematically studied the spectrum-effect-toxic relationship of ER,and initially defined its efficacy/toxic substance basis.This article intends to establish a research method for the use of traditional Chinese medicine of "the older,the better" based on accelerated test.The various indicators were integerated to comprehensively evaluate the quality changes in the process of aging of traditional Chinese medicine,including external properties,components,efficacy/toxicity,etc.Then,the aging process of ER who has the samilar source was investigated,the dynamic change rule of the active ingredient group was traced,and the balance point between efficacy and toxicity was evalued.Finally,the scientific connotation of "the older,the better" was elucidated.Methods: 1.In order to systematically explore the changes in the chemical composition of ER during the aging process,it was divided into three parts: alcohol extract,water extract and volatile oil.Qualitative fingerprint analysis and multi-index quantification of alcohol extracts and aqueous extracts of ER were performed by high performance liquid chromatography(HPLC);Fingerprint characterization and Quantification by normalization of volatile constituents were carried out by gas chromatography-mass spectrometry(GC-MS).2.The ER with the similar source was divided into normal aging group,accelerated aging group and frozen aging group,which were stored at room temperature,37 ? and-18 ?.They were sampled according to the experimental design time point,and analyzed by the above analysis method.The changes of appearance,volatile oil content,components and their contents of each group were recorded,and the dynamic changes of the above indicators in the aging stage were tracked to evaluate the quality changes during the aging process.3.The typical acetic acid twist and hot plate method in mice were adopted.The pharmacological differences between the initial aging(Week 0)with the key points who have significant changes were evalued.4.Using the Step Thermostatic Method,several sets of higher temperature acceleration experiments were set up,and the reaction constants of the respective compounds were obtained according to the Arrhenius Formula,and the time conversion coefficient ? was extrapolated.5.Combining the research results of all groups,and evaluating the ER's quality from the following aspects such as the external appearance,chemical composition(fingerprint,active ingredient group),efficacy(analgesia,vomiting)and toxicity(acute toxicity,hepatotoxicity),etc.In different aging process,the scientific connotation of ER's "the older,the better" was systematically eludcidated based on the theory of traditional Chinese medicine.Restult: 1.The fingerprint methods of ER's ethanol extracts and water extracts were established.In ethanol extracts 33 common peaks were listed and 23 of them were identified.In water extracts 27 common peaks were listed and 17 of them were identified.The quantitative methods for 8 and 7 active components in ER's ethanol extracts and water extracts were established.Meanwhile 51 components in volatile oil of ER were identified with the GC-MS method,and quantified with normalization method.2.All the herbs were in accordance with the requirements of the China Pharmacopoeia,and their quality did not change significantly during the research process.(1)In terms of external appearance,the color of ER's fruits and extracts gradually deepens and the pungent odor was weakened.(2)Qualitative and semi-quantitative fingerprint analysis indicate: In the ethanol extracts,the peak areas of trans-caffeoylgluconic acid,cryptochlorogenic acid,caffeic acid,trans-caffeoyl gluconate-?-lactone,13,15 and 18 increased;the peak areas of other organic acids,isorhamnetin-3-O-rutinoside,6,7,8 and 22 decreased;the peak areas of quinolones alkaloids,14 and 24 remained stable.In the water extracts,the peak area of caffeic acid,trans-caffeoyl gluconate-?-lactone,15 and 19 remained stable;the peak area of other organic acids,isorhamnetin-3-O-rutinoside,6,7,8,11,14,16,17,and 18decreased.(3)In terms of quantitative results,the changes trends of alcohol extracts and water extracts were consistent,the contents of chlorogenic acid decreased;the contents of dehydrogenated and rutaecarpine increased;and the contents of evodiamine and dihydroevocarpine showed no significant changes.In the accelerated aging group,the contents of rutin and hyperoside decreased,and quercetin was grow out of nothing,but no significant changes were found in the normal aging group.(4)In terms of volatile oil,the amounts of volatile oil diminished significantly and the percentage of ?-ocimene and ?-ocimene decreased.(5)During the aging process,the above changes showed a trend of quick to slow.At the week 76,those changes became significantly slower and seemingly entered the platform period.3.Compared with the blank group,all the experimental group showed marked analgesic effect.The analgesic effect of alcohol extracts were better than water extracts.And there were no significant differences in the analgesic effect between the initial group(week 0),the normal group(week 76),and the accelerated aging group(week 76),which indicated that there were no significant changes in the analgesic effect of ER before and after the aging,so the week 76 was not balance point.4.By using the Step Thermostatic Method,we obtained the time conversion coefficient ?=1.97,which means that accelerated group aging(37 ?)for 1 year is equivalent to normal group aging(20 ?)for 1.97 years.Conclusion: This article established a research method based on accelerated test,which was used to study the theory of traditional Chinese medicine of "the older,the better".In this work,we designed the normal aging group,accelerated aging group and freezing aging group.The three groups can compensated and verified each other,thus the comparability and reliability of the research results can be guaranteed.Under the studied conditions,the conversion coefficient ?=1.97.The results showed that it was reasonable and feasible to use the acceleration test of simulating the natural environment to study the "the older,the better" of Chinese medicine.During the aging process,ER's quality was stable,toxicity and dryness were decreased.Therefore,"the older,the better" has its scientific connations.After the aging,ER's functional ingredients such as rutaecarpine and dehydroevodiamine increased slightly,the dryness and toxicity ingredients such as volatile oil decreased significantly.That was the main internal reason of ER's "the older,the better".ER may have the best aging process,the study of the best aging time is still under the way.