Herbalogical study on historical evolution of Juhong and Huajuhong / 中国中药杂志

In ancient times, there were two types of "Juhong" came from the tangerines(Citrus reticulata) and the pomelos(C. grandis and its cultivars), which corresponded to Juhong and Huajuhong recorded in the Chinese Pharmacopoeia respectively. In different periods, Juhong basically came from the same species and the same medicinal parts, but there were also some differences. This article sorted out the ancient and modern literature, under the guidance of "Succession theory of Medicinal materials varieties" and "Change theory of Medicinal materials varieties"(XIE Zong-wan), and combined with field investigation, the evolution and reasons of the original plants and medicinal parts of Juhong were analyzed. In the Han Dynasty and before, the peel of tangerines and pomelos were both used as medicine. In the Southern and Northern Dynasties, the way tangerine peel was used was dried and aged, and then "soaked in hot water and scraped off the mesocarp", which had the essence of only using exocarp as medicine of Juhong already, and its original plant was C. reticalata. In the Song Dynasty, the name of "Juhong" and its medicinal usage were recorded in book on materia medica, and the species and medicinal parts of tangerine were inherited from the previous dynasties. The way tangerine peel was used was only dried and aged without removing the mesocarp. The medicinal material obtained by the way was called Chenpi(dried and aged tangerine peel). The item "Juhong" listing as a separate medicinal material was first recorded in the Collected Discussions from Materia Medica(Bencao Huiyan) in the Ming Dynasty. In the Ming Dynasty, the Dao-di habitat of Juhong was recorded as Guangdong province in most books on materia medica, and the original plants probably were C. reticalata and C. grandis 'Tomentosa'(Huazhou pomelo, a special cultivated species of C. grandis produced in Huazhou, Guangdong, which was recorded in the Chinese Pharmacopoeia as "Huajuhong"), according to the records in the local chronicles. During the Qing Dynasty and the Republic of China, the original plants of Juhong were C. reticalata and C. grandis 'Tomentosa'. Of the two, the latter one was considered as the better. As far the medicinal part, it was still the exocarp, while the whole young fruit of C. grandis 'Tomentosa' began to be used as medicine. After the founding of The People's Republic of China, the exocarps of Citrus reticalata, C. grandis and C. grandis 'Tomentosa' were listed in the Chinese Pharmacopoeia under "Juhong". From the Northern and Southern Dynasties to the Republic of China, C. grandis exocarp was a fake of Juhong. Therefore, it was contradictory to historical records that C. grandis exocarp was listed in the Chinese Pharmacopoeia as Huajuhong. Juhong had been divided into two types as "Juhong" and "Huajuhong" since 1985. The medicinal part of Huajuhong was only the exocarp of immature and nearly mature fruits, but not the whole young fruit, the actual mainstream medicinal part of Huajuhong. The results are helpful to clarify the historical evolution of species and medicinal parts of Juhong and Huajuhong. It is suggested that in the next edition of Chinese Pharmacopoeia, only C. grandis 'Tomentosa' should be included as the original plant of Huajuhong, and C. grandis should be deleted, and the young fruit should be added in the medicinal parts besides the exocarp of immature and nearly mature fruit.

Common mechanism of Citrus Grandis Exocarpium in treatment of chronic obstructive pulmonary disease and lung cancer / 中草药·英文版

Objective: “Same treatment for different diseases” is a unique treatment strategy in traditional Chinese medicine. Two kinds of malignant respiratory diseases endanger human health-chronic obstructive pulmonary disease (COPD) and lung cancer. Citrus Grandis Exocarpium (Huajuhong in Chinese, HJH), a famous herbal, is always applied by Chinese medicine practitioners to dispersion the lung to resolve phlegm based on “syndrome differentiation and treatment” theory. However, the common mechanism for HJH's treatment of COPD and lung cancer is not clear. Methods: In this study, based on network pharmacology and molecular docking technology, the common mechanism of HJH in the treatment of COPD and lung cancer was studied. The active ingredients and related targets of HJH were integrated from TCMSP, BATMAN-TAM, STP, and Pubchem databases. The standard names of these targets were united by UniProt database. Targets of COPD and lung cancer were enriched through GeneCards, NCBI (Gene), Therapeutic Target Database, and DisGeNET (v7.0) databases. Then the intersection targets of HJH and diseases were obtained. The STRING network and the Cytoscape 3.7.2 were used to construct PPI network, the DAVID database was used to perform GO and KEGG analysis. Then Cytoscape 3.6.1 was used to build “ingredient-target-signal pathway” network. Finally, AutoDock 1.5.6 software was used to perform molecular docking of key proteins and molecules. Results: Eleven active ingredients in HJH were obtained by searching the database, corresponding to 184 HJH-COPD-lung cancer targets intersection. The results of biological network analysis showed that naringenin, the active component in HJH, could mainly act on target proteins such as AKT1, EGFR. Then through positive regulation of vasoconstriction and other biological processes, naringenin could regulate estrogen signaling pathway, VEGF signaling pathway, HIF-1 signaling pathway, ErbB signaling pathway, PI3K-Akt signaling pathway to play an important role in the treatment of both COPD and lung cancer. Conclusion: Network pharmacology was employed to systematically investigate the active ingredients and targets of HJH in treatment of COPD and lung cancer. And then, the common pharmacodynamic network of HJH for the two malignant respiratory diseases was firstly described. Furthermore, naringenin was proved to strongly bind with AKT1 and EGFR. It may provide the scientific basis for understanding the “Same treatment for different diseases” strategy in traditional Chinese medicine and inspirit subsequent drug discovery for COPD, lung cancer and other malignant lung diseases.

Identification and evaluation of Citrus grandis based on DNA barcode,UPLC and chromaticity method / 中国中药杂志

In order to identify the source of Citrus grandis and evaluate its quality originate from two areas comprehensively,DNA barcode was used to identify 26 samples of C. grandis. The content of naringin,rhoifolin,naringenin and apigenin was determined by UPLC method,and the color difference was numerically studied by color difference analyzer,which was related to the effective components of C. grandis. The results showed that samples was the source of C. grandis in both regions. The ITS2 sequence length was about400-500 bp,and the sequence similarity reached 99. 82%. There was only one base deletion in the two groups. There was one base A in some medicinal materials of Guangdong at 330 bp,but no base in Chongqing. The contents of naringin and rhoifolin in Chongqing samples were higher than those in Guangdong samples,and there were statistical differences between naringenin and apigenin. The chroma value showed that L*value of Guangdong was larger,a*value was smaller,L*value of Chongqing was smaller,and a*value was larger,while the b*value of both was not significantly different; The results of correlation analysis showed that naringin,rhoifolin,naringenin were positively correlated with L*,b*value,negatively correlated with a*value,and apigenin had no correlation with L*,a*,b*value. In this study,the scientific identification and evaluation of C. grandis was carried out to provide a new idea for the further study of the rapid identification and evaluation of C. grandis.

Determination of 106 Pesticide Residues in Citrus Grandis Exocarpium / 中国药学杂志

OBJECTIVE: To establish a method for determination of pesticide residues in Citrus Grandis Exocarpium by mass spectrometry. METHODS: After vortex extraction with acetonitrile, the samples were purified with prime-HLB SPE. GC-MS/MS analysis was performed on a DB17 MS(30 m×0.25 mm, 0.25 μm)capillary column with electron impact(EI) source. LC-MS/MS analysis was performed on a Phenomenex Kinetex C18(2.1 mm×100 mm, 2.6 μm) column with gradient elution using water(containing 5 mmol•L-1 ammonium formate and 0.1% formic acid) and 95% acetonitrile(containing 5 mmol•L-1 ammonium formate and 0.1% formic acid). Electrospray ionization(ESI) source was applied by positive ionization in multiple reaction monitoring(MRM) mode. RESULTS: The calibration curves of 106 pesticide residues showed good linearity with correlation coefficients above 0.990 9. The average recoveries for all target compounds fell in the range of 57.6%-134.4%, with RSDs below 15%. Seven kinds of prohibited pesticides and 25 strictly restricted and commonly used ones were detected in 20 batches of samples, which mainly belong to insecticide, acaricide and fungicide. CONCLUSION: The method is fast, accurate and highly sensitive for the screening and confirmation of pesticide residues in Citrus Grandis Exocarpium, which provides a basis for the control of pesticide residues in similar mattrix medicinal plants.

Comparison of content of five flavones in Citrus grandis “Tomentosa” and Citrus grandis with quantitative analysis of muli-components by single marker / 中草药

Objective To establish quantitative analysis of multi-components by single marker (QAMS) to compare five flavones in Citrus grandis “Tomentosa” and C. grandis. Methods Four relative correction factors (RCFs) of neohesperidin, rhoifolin, naringenin, apigenin were established in the HPLC method with the Naringin as internal standard, which were used to determine the content of four flavones in C. grandis “Tomentosa” and C. grandis. Meanwhile, external standard method (ESM) was employed to calculate the content of five flavones. The difference between ESM and QAMS were analyzed to evaluate the accuracy of QAMS. T-test was used to compare five flavones in C. grandis ‘Tomentosa’ and C. grandis. Results RCFs of neohesperidin, rhoifolin, naringenin, apigenin were 0.898, 1.519, 0.313, 0.406, and repeatability was good in different experimental conditions (RSD < 3.0 %). There were no significant differences in the quantitative analysis results of two methods. Content of naringin, rhoifolin and naringenin in two species were significant different. Conclusion The RCFs of neohesperidin, rhoifolin, naringenin, apigenin as reference to naringin are accurate and feasible. Content of flavones in Citrus grandis “Tomentosa” and C. grandis are obvious different. QASM can be applied as a strategy for quality control of C. grandis.

Isolation and identification of chemical constituents from peels of Citrus changshan-huyou Y.B.Chang / 北京大学学报(医学版)

Objective:To study the chemical constituents of the peels collected from Citrus changshan-huyou Y.B.Chang,and further screen the bioactive components as the lead structures.Methods: These compounds were isolated by repeated flash column chromatography on silica gel and Sephadex LH-20.The structures of isolated compounds were elucidated by using IR,EIMS,and NMR analyses.Results: One novel compound along with nine known compounds were obtained and identified as huyoujiasu(Ⅰ),3-hydroxy-4-methoxybenzoic acid(Ⅱ),3,4-dihydroxybenzoic acid(Ⅲ),fatty acid(Ⅳ),gly-ceride(Ⅴ),6,7-dimethoxycoumarin(Ⅵ),6′,7′-dihydroxybergamottin(Ⅶ),daucossterol A(Ⅷ),daucossterol B(Ⅸ) and huyou-triterpenoid(Ⅹ).Conclusion: One new compound,Huyoujiasu,was obtained from these peels by repeated column chromatography,and the other known compounds were isolated from this fruit for the first time as well.

Observation on Dynamic Changes of Flavonoids Contents of Citrus grandis(L.)Osbeck var.tomentosa Hort / 广州中医药大学学报

【Objective】To observe the dynamic changes of flavonoids contents of Citrus grandis(L.)Osbeck var.tomentosa Hort..【Methods】The total flavonoid content was determined by spectral photometric analysis and naringin content by HPLC,and the fingerprints of Citrus grandis(L.)Osbeck var.tomentosa Hort.were studied by HPLC.【Results】With the increase of the fruit age,total flavonoid contents in the peel and the leaves of Citrus grandis(L.)Osbeck var.tomentosa Hort.decreased obviously.Fingerprints results showed that at the early fruit age the rhoifolin content in the peel increased with the fruit age and began to decrease 62 days later,while the rhoifolin content in the leaves showed no changes.【Conclusion】In terms of active components contents and economic value,the optimal collecting time for Citrus grandis(L.)Osbeck var.tomentosa Hort.is:young fruit,34 days,and immature fruit,55 days.

Dynamic Variation of Flavonoids Contents in Flowers of Citrus grandis at Different Flowering Periods / 中药新药与临床药理

Objective To investigate the dynamic variation of flavonoids contents in the flowers of Citrus grandis at different flowering periods. Methods The content of total flavonoids in the flowers of Citrus grandis was determined by means of ultraviolet-visible absorption spectroscopy ,and the contents of naringin and rhoifolin were determined by HPLC. Results The content of total flavonoids in the flowers during the flower withering period ,the young fruit period ,the blossom period,and the budding period was 306.90 mg.g-1,277.93 mg.g-1,215.55 mg.g-1 and 162.74 mg.g-1 respectively. The naringin content during the above four different periods was 246.31 mg.g-1,213.93 mg.g-1,175.94 mg.g-1 and 130.90 mg.g-1 respectively. The rhoifolin content during the four periods was unchanged. Conclusion The contents of total flavonoids and naringin in flowers of Citrus grandis during flower withering period are the highest.

Comparative Study on the Content of Total Flavonoid and Naringin in Different Cultivars of Citrus Grandis ‘Tomentosa’ / 中药新药与临床药理

Objective To choose a best cultivar by comparing the content of total flavonoid and naringin between different cultivars of Citrus grandis 'tomentosa'.Methods The content of total flavonoids was determined by spetral photometric analysis.The content of naringin was determined by HPLC.Results The content of total flavonoid was 19.24 % and 18.06 %,and naringin content was 11.72 % and 12.38 % in the cultivars from Dachaling and Fengwei respectively,which were much higher than the other cultivars.Conclusion The contents of total flavonoid and naringin were different in different cultivars of Citrus grandis 'tomentosa'.The cultivars of Citrus grandis 'tomentosa' from Dachaling and Fengwei have the best quality.

Comparison of Ultrasonic Extraction and Soxhlet Extraction on the Content Dete rmination of Naringin in Citrus grandis / 中药新药与临床药理

Objective To establish the optimal extracted method for content dete rmination of naringin in Citrus grandis. Methods RP-HPLC was used to determinat e the content of naringin extracted with the above two methods from different ye ar samples of Citrus grandis. Results The average content extracted with ultraso nic extraction was 13.53 %,and the average content extracted with soxhlet extr action was 11.98 %,there being insignificant difference between the two method s. Conclusion The content of naringin extracted with ultrasonic extraction is mo re than that with the soxhlet extraction,which be receipted in Chinese pharmeco pia. And ultrasonic extraction method is more convenient and can save time.

Separation and purification of naringin from Citrus grandis Tomentosa by macroporous adsorption resin / 中成药

AIM: To optimize the macroporous resin for separating naringin. METHODS: Putting the extract fluid of Citrus grandis Tomentosa into pillar was adsorbed with macroporous resin, then washed by alcohol with the different concentration in succession to determine the content of naringin by HPLC. RESULTS: The six macroporous resins' effects differ greatly. CONCLUSION: HPD450 macroporous resin is effective to separate the naringin.