Study On The Screening Of Characteristic Components Of Agarwood And The Comparison Of Pharmacodynamics

Agarwood,a resinous heartwood of Aquilaria sinensis(Lour.),possesses the actions of regulating qi and relieving pain,warming and stopping vomiting,absorbing qi and relieving asthma.At present,domestic agarwood resources are mainly distributed in the coastal areas of Guangdong,Guangxi and Hainan.Due to years of predatory logging,destruction of forest resources and low yields,the wild resources of agarwood have been drastically reduced.To meet the increasing demand of the market,artificial planting and regulation of inducing technique has become the main way to solve the resources of agarwood.However,the quality of agarwood obtained by different production methods is uneven.Different growth patterns and induced methods may affect the content and composition of secondary metabolites,especially the accumulation of active compounds,which leads to the difference in the therapeutic efficacy of agarwood from different production methods.Literature reports indicate that the active compounds of this medicinal material mainly include volatile oils and 2-(2-phenylethyl)chromones.In the clinical application and production process of agarwood,how to effectively combine with its specific pharmacological efficacy and accurately identify the main secondary metabolic chemical components,to determine the chemical components of different production methods of agarwood is the key issue.In this study,we analyzed the differences in the overall chemical composition characteristics of agarwood from different sources.The content of total 2-(2-phenylethyl)chromone and agarotetrol were determined by UV-Vis and HPLC techniques to clarify the quality differences of agarwood from different production methods.Based on GC-MS and UPLC-Q-Exactive Orbitrap/MS metabolomics technology to identify the non-volatile and volatile components of agarwood,At the same time,the OPLS-DA method was used to screen and identify marker compounds with different qualities.And study the anti-inflammatory and analgesic effects of agarwood from different sources,and to compare the differences in pharmacodynamics.The main results are as follows:(1)Near-infrared spectroscopy showed that the total metabolic components of agarwood produced by different methods were similar,and principal component analysis(PCA)could be used to effectively identify them,indicating that there were differences in content accumulation among different production methods.The determination results of two main characteristic components showed that the content of total 2-(2-phenylethyl)chromone was between 3.58±0.11～22.56±0.22%,and the agarotetrol was between 2.21±0.03～13.71±0.06mg/g.The quality difference of the medicinal materials of agarwood with different production methods was confirmed.From the perspective of chemical composition content,it is speculated that the quality of wild agarwood is better than cultivated agarwood.(2)Based on the metabolomics technology of GC-MS,the metabolic profile of the volatile oil components of agarwood obtained from different growth methods and induced methods is studied.The NIST 14 standard database and linear retention index were used to qualitatively identify volatile components.A total of 76 volatile chemical components were identified,including sesquiterpenes,chromones,aromatics,and triterpenes.An effective identification model was established based on OPLS-DA,and the volatile characteristic difference chemical components of 11 wild and cultivated agarwood and 6 cultivated agarwood were successfully selected according to the importance projected value(VIP).(3)Based on the metabolomics technology of UPLC-Q-Exactive Orbitrap/MS,the metabolic profile of the chromones of agarwood obtained from different growth methods and induced methods is studied.Matched with self-built agarwood chromone database and some reference substances,a total of 79 2-(2-phenethyl)chromone compounds were identified.They include 2-(2-phenylethyl)chromone type(PECs),5,6,7,8-tetrahydro-2-(2-phenylethyl)chromones type(THPECs),5,6-epoxy-2-(2-phenylethyl)chromones types(EPECs)and 5,6:7,8-diepoxy-2-(2-phenylethyl)chromones types(DEPECs).An effective identification model was established based on OPLS-DA,and the non-volatile characteristic difference chemical components of20 wild and cultivated agarwood and 14 cultivated agarwood were successfully selected according to the importance projected value(VIP).(4)According to the analysis of the metabolic characteristics of agarwood from different production methods,the acetic acid-induced writhing test in mice and xyleneinduced mouse ear swelling were used to investigate the anti-inflammatory and analgesic effects of agarwood induced by different methods.The experimental results show that the agarwood induced by the natural injuries shows the strongest antiinflammatory and analgesic effect.Among the agarwood induced by the artificial method,the agarwood induced by burning-chisel-drilling shows the best antiinflammatory effect,and the agarwood induced by holing shows the best analgesic effect.