Comparison Of Components Of Zanthoxylum Armatum DC. From Different Areas And Study On The Changes Of Chemical Components During Storage

Zanthoxyli Armati Pericarpium stems from the dried and ripe peel of Zanthoxylum armatum DC.It is pungent and warm in nature.It has the effects of relieving pain,killing insects and itching.It is also known as"TENG JIAO","WAN HUA ZHEN","BAI ZONG GUAN","ZHU YE ZONG GUAN","SHAN HUA JIAO",etc.It is included in the local standards of Standard of Chinese Medicinal Materials and Decoction Pieces Prepared by Sichuan Provincial Drug Administration（attached Edition of 2020 edition）,Hunan provincial standard for Chinese herbal medicine（2009Edition）and Guangxi provincial standard for Chinese herbal medicine（1990 edition）.Zanthoxylum armatum DC.is widely planted in Chongqing,Sichuan and other places.The research shows that there are also significant differences in the main volatile components and percentage content of Zanthoxylum armatum DC.from different producing areas,but the research on the difference of multi-component content and antioxidant activity from different producing areas has not been reported in the literature.Therefore,taking Zanthoxylum armatum DC.from Liangshan,Bazhong and Chongqing as research samples,this paper studies the differences of color value,volatile oil,flavonoids,amide content and antioxidant activity of Zanthoxylum armatum DC.from different producing areas,and analyzes the correlation between color value,chemical composition and antioxidant activity,so as to find out the indexes with high correlation with antioxidant activity of Zanthoxylum armatum DC.,The entropy weight TOPSIS multi index comprehensive evaluation model is established to improve the existing standards.The changes of water,chemical composition and content of Zanthoxylum armatum DC.during storage were studied to provide experimental basis for reasonable storage and maintenance of Zanthoxylum armatum DC..The specific research contents are as follows:1 Comparative study on the components of Zanthoxylum armatum DC.from different producing areas1.1 Comparative study on appearance colorIt is reported in the literature that the color of traditional Chinese medicine is one of the important indexes of the quality of traditional Chinese medicine.In this paper,the powder color of 27 batches of Zanthoxylum armatum DC.from Liangshan,Bazhong and Chongqing was objectively quantified by spectrophotometer.Comparing the color values of different producing areas,it was found that the color of Zanthoxylum armatum DC.samples from different producing areas was different,and the color of Zanthoxylum armatum DC.from Liangshan Prefecture was dark and yellow green.In addition,the cluster analysis was carried out by using IBM SPSS statistics 21 data analysis software.The results showed that the samples of Zanthoxylum armatum DC.in Liangshan Prefecture were basically clustered into one class,and the samples from other producing areas were clustered into one class.1.2 comparative study on composition of volatile oilVolatile oil is the main component of the aroma of Zanthoxylum armatum DC..In this paper,the volatile oil of Zanthoxylum armatum DC.from different producing areas was extracted by steam distillation.The volatile components of Zanthoxylum armatum DC.were qualitatively analyzed by gas chromatography-mass spectrometry.A total of63 compounds were isolated and identified,accounting for 99.06%～99.99%of the total components,including olefins,alcohols,aldehydes,ketones,hydrocarbons,etc Alcohols are the main components,alkenes are the most abundant components of volatile oil,and alcohols are the components with the highest relative content in volatile oil.The identified compounds linalool,d-limonene and sabinene are common components,among which linalool（83.95%）accounts for a relatively high percentage in Zanthoxylum armatum DC.in Daying county（S13）of Suining,d-limonene（15.34%）Sabinene（9.49%）accounted for a high percentage of Zanthoxylum armatum DC.in Dechang County（S4）of Liangshan Prefecture.1.3 comparative study on the content of flavonoidsIn this paper,the contents of four flavonoids（rutin,hypericin,isoquercetin and quercetin）of Zanthoxylum armatum DC.from different areas were determined by HPLC,and the total flavonoids were determined by spectrophotometry.The results showed that the total flavonoids content ranged from 30.460 7 to 90.173 1 mg·g-1.The total flavonoids content in Tongjiang County（S12）of Bazhong City was 3 times of the content of total flavonoids in Jiajiang County（S14）of Leshan City,and the content of hypericin ranges from 0.4427 to 3.9832 mg·g-1.The content of hypericin in Yanyuan County（S3）of Liangshan Prefecture is nearly 10 times higher than that in Butuo county（S8）of Liangshan Prefecture.There are great differences in the content of total flavonoids and four flavonoids from different producing areas.1.4 comparative study on the content of amidesIn this paper,high performance liquid chromatography was used to determine hydroxyl-α-capsaicin,hydroxyl-β-capsaicin,hydroxyl-γ-capsaicin of Zanthoxylum armatum DC.from different area.The results showed that the total amide content of of Zanthoxylum armatum DC.from different areas was 55.572 9～174.700 3 mg/g,the content of hydroxyl-α-capsaicin is 21.443～152.8664 mg/g,the content of hydroxyl-β-capsaicin is 0～21.832 4 mg/g,and the content of TL is 24.624 7～173.958 8mg/g.There are great differences in the content of amide among different producing areas.2 Study on antioxidant components of Zanthoxylum armatum DC.2.1 Comparison of antioxidant activity of Zanthoxylum armatum DC.from different producing areasThe antioxidant activity of Zanthoxylum armatum DC.from different areas was analyzed by DPPH and ABTS free radical scavenging test,and the correlation between flavonoids and antioxidant activity was analyzed by Pearson bivariate correlation analysis.The results showed that there were great differences in antioxidant activity of Zanthoxylum armatum DC.from different producing areas.Bazhong Tongjiang（S12）has the highest scavenging capacity for DPPH free radicals.Compared with other regions,the three batches of methanol extracts of Zanthoxylum armatum DC.from Bazhong have higher scavenging capacity for DPPH,which is consistent with the determination results of flavonoids,but Mianyang Santai has the best scavenging capacity for ABTS free radicals,indicating that there are differences in the results of different antioxidant activity tests in vitro.2.2 Correlation study on appearance color,chemical composition and antioxidant activity of Zanthoxylum armatum DC.The correlation between the content of flavonoids and color value in Zanthoxylum armatum DC.from different producing areas was studied by using IBM SPSS statistics21 software.It was found that the higher the content of hypericin and isoquercetin,the darker the color of Zanthoxylum armatum DC.was red,the higher the content of total flavonoids,and the darker the color of Zanthoxylum armatum DC.was green.Pearson bivariate correlation analysis was used to analyze the correlation between flavonoids and antioxidant activity.It was found that there was a significant negative correlation between total flavonoids and rutin and the EC₅₀ values of DPPH and ABTS free radical clearance,indicating that total flavonoids and rutin can be used as one of the indicators to evaluate the antioxidant capacity of Zanthoxylum armatum DC..The correlation coefficient between rutin content and total flavonoids content was high（correlation coefficient≥0.6）,and there was high consistency.3 Establishment of TOPSIS multi index comprehensive evaluation model with entropy weight of Zanthoxylum armatum DC.The mathematical model established by TOPSIS method can realize multi index component comprehensive analysis by approaching the ideal solution,which is in line with the characteristics of multi-component and multi-target action of traditional Chinese medicine.Entropy weight method gives weight coefficient according to the correlation between the content and function of each index component,which can more accurately evaluate the quality of traditional Chinese medicine.Therefore,the combination of entropy weight method and TOPSIS method can objectively and accurately reflect the contribution of each index to the quality of Zanthoxylum armatum DC.By establishing the entropy weight TOPSIS multi index comprehensive evaluation model,it is found that the relative proximity of 15 batches of Zanthoxylum armatum DC.is between 0.287～0.611,indicating that there are some differences in the quality of each batch of Zanthoxylum armatum DC.Through cluster analysis,it is found that those with relative proximity>0.53 are basically clustered into one class,and those with relative proximity<0.53 are basically clustered into one class.In addition,linalool,sabinene,d-limonene,laurene and TL,total amides and hydroxyl-α-capsaicin,hydroxyl-β-capsaicin in the samples are high relative proximity to each batch of Zanthoxylum armatum DC.The content of these is also relatively high,and the weight coefficient is large.Hydroxyl-α-capsaicin,hydroxyl-β-capsaicin is the main hemp flavor substance in Zanthoxylum armatum DC..Therefore,this study suggests that the content of hydroxyl-α-capsaicin and hydroxyl-β-capsaicin was included in the quality evaluation index of Zanthoxylum armatum DC.,so as to lay a foundation for the improvement of its quality standard.4 Study on the changes of components of Zanthoxylum armatum DC.during storage4.1 Study on changes of water content and water activityIn this paper,the water content and water activity of Zanthoxylum armatum DC.under different storage time were measured by toluene method and water activity,which showed an upward trend.4.2 Study on the content change of volatile oilThe composition and content of volatile oil in Zanthoxylum armatum DC.during storage were determined by gas chromatography-mass spectrometry.The results showed that the content of volatile oil in Zanthoxylum armatum DC.decreased with the extension of storage time.The relative content of alkene components represented by monoterpene components such as laurene and juniper decreased gradually,and the relative content of alcohol components represented by linalool increased gradually.Except that the relative content of main components of volatile oil changed,its trace components may have oxidative deterioration.During storage,new components and some components disappeared in three batches of Zanthoxylum armatum DC.4.3 Study on the content change of flavonoidsIn this paper,the content changes of three flavonoids（rutin,hypericin and isoquercetin）in Zanthoxylum armatum DC.during storage were determined by HPLC,and the content of total flavonoids was determined by spectrophotometry and aluminum nitrate sodium nitrite color method.The results showed that the content of flavonoids in Zanthoxylum armatum DC.showed a gradual downward trend during storage,Especially in the previous three months,the downward trend was obvious,and then gradually stabilized.4.4 Study on content change of amidesThe amide components and contents were determined by HPLC and spectrophotometry.The results showed that with the extension of storage time,the content of total amide of Zanthoxylum armatum DC.was decreased.The content of hydroxyl-α-capsaicin showed a downward trend,decreased significantly within 3months before storage,and began to fluctuate after reducing to a certain extent.In conclusion,this paper investigated the changes of water,volatile oil,flavonoids and amides in the storage of Zanthoxylum armatum DC..The results showed that the water content in Zanthoxylum armatum DC gradually increased with the extension of storage time,suggesting that it should be sealed during storage.The changes of volatile oil,amide and other unstable chemical components and their contents suggest that the storage of Zanthoxylum armatum DC should be protected from light and low temperature.