AI-driven determination of active compounds and investigation of multi-pharmacological effects of Chrysanthemi Flos.

BACKGROUND: Chrysanthemi Flos as a medicine food homology species is widely used in the prevention and treatment of diseases, whereas comprehensive research of its active compounds related to multi-pharmacological effects remains limited. This study aimed to systematically explore the active compounds through artificial intelligence-based target prediction and activity evaluation. METHODS: The information on compounds in Chrysanthemi Flos was obtained from six cultivars containing Gongju, Chuju, Huaiju, Boju, Hangbaiju, and Fubaiju, using UPLC-Q-TOF/MS. The main differential metabolites in six cultivars were also screened through the PLS-DA model. Then the potential targets of differential compounds were predicted via the DrugBAN model. Enrichment and topological analysis of compound-target networks were performed to identify key pharmaceutical compounds. Subsequently, the pharmacological effects of predictively active compounds were confirmed in vitro. Based on the active compounds, the pharmacological activities of Chrysanthemi Flos from the six origins were also investigated and compared for the further evaluation of medicinal quality. RESULTS: A total of 155 secondary metabolites were obtained from Chrysanthemi Flos. Among them, 26 differential components were screened, and 9 key pharmacological compounds with 1141 targets were identified. Enrichment analysis indicated the main pharmacological effects of Chrysanthemi Flos related to inflammation, oxidative stress, and lipid metabolism. In addition, 9 key pharmaceutical compounds were evaluated in vitro experiments, indicating the significant therapeutic effect in regulating inflammation, oxidative stress, and lipid metabolism. CONCLUSION: This study successfully identified 9 key pharmaceutical compounds in Chrysanthemi Flos and predicted the pharmacodynamic advantages of six origins. The findings would provide improved guidance for the discovery of active constituents and the assessment of pharmacodynamic advantages of different geographical origins.

Strategies for the quality control of Chrysanthemi Flos: Rapid quantification and end-to-end fingerprint conversion based on FT-NIR spectroscopy.

INTRODUCTION: Chrysanthemi Flos (CF) is widely used as a natural medicine or tea. Due to its diverse cultivation regions, CF exhibits varying quality. Therefore, the quality and swiftness in evaluation holds paramount significance for CF. OBJECTIVE: The aim of the study was to construct a comprehensive evaluation strategy for assessing CF quality using HPLC, near-infrared (NIR) spectroscopy, and chemometrics, which included the rapid quantification analyses of chemical components and the Fourier transform (FT)-NIR to HPLC conversion of fingerprints. MATERIALS AND METHODS: A total of 145 CF samples were utilised for data collection via NIR spectroscopy and HPLC. The partial least squares regression (PLSR) models were optimised using various spectral preprocessing and variable selection methods to predict the chemical composition content in CF. Both direct standardisation (DS) and PLSR algorithms were employed to establish the fingerprint conversion model from the FT-NIR spectrum to HPLC, and the model's performance was assessed through similarity and cluster analysis. RESULTS: The optimised PLSR quantitative models can effectively predict the content of eight chemical components in CF. Both DS and PLSR algorithms achieve the calibration conversion of CF fingerprints from FT-NIR to HPLC, and the predicted and measured HPLC fingerprints are highly similar. Notably, the best model relies on CF powder FT-NIR spectra and DS algorithm [root mean square error of prediction (RMSEP) = 2.7590, R2 = 0.8558]. A high average similarity (0.9184) prevails between predicted and measured fingerprints of test set samples, and the results of the clustering analysis exhibit a high level of consistency. CONCLUSION: This comprehensive strategy provides a novel and dependable approach for the rapid quality evaluation of CF.

Identification of the anti-inflammatory quality markers of Chrysanthemi Flos (Juhua) using spectrum-effect relationships combined with bioactivity re-evaluation.

Chrysanthemi Flos (Juhua), an edible herbal medicine that possesses efficacies of dispersing wind, clearing heat and detoxifying. Studies have demonstrated that the health benefits of Chrysanthemi Flos are largely attributable to its anti-inflammatory effects. However, the correlation between the compounds monitored by the current quality control methods and the anti-inflammatory effects of Chrysanthemi Flos is unclear. In order to better control the quality of Chrysanthemi Flos, the identification of anti-inflammatory quality markers (Q-markers) of Chrysanthemi Flos was performed. The chemical components of Chrysanthemi Flos were profiled by HPLC fingerprints combined with chemometrics methods. Simultaneously, the anti-inflammatory activities of 10 batches of water extracts of Chrysanthemi Flos were evaluated in lipopolysaccharide-activated RAW 264.7 macrophages cells. Gray correlation analysis was performed to assess the relationship between the anti-inflammatory activity and chemical properties. The results showed that 13 common peaks were closely correlated with the anti-inflammatory effect, and further bioactivity re-evaluation confirmed that 10 known compounds exerted a strong anti-inflammatory effect. The quantitative analysis of the 10 Q-markers showed that the 25 batches of samples could be discriminated into different zones according to their producing areas. Conclusively, the present work identified 10 anti-inflammatory Q-markers of Chrysanthemi Flos using spectrum-effect relationships combined with bioactivity re-evaluation.

Herbal Textual Research on Chrysanthemi Flos in Famous Classical Formulas / 中国实验方剂学杂志

To conduct textual research on the records of Chrysanthemi Flos in the ancient literature from the aspects of name， origin， species， scientific name， origin， harvesting， processing， quality and so on， and the modern literature was sorted out to clarify the relationship between the ancient and modern times， so as to provide a reference for the development of famous classical formulas containing Chrysanthemi Flos. Chrysanthemi Flos is an ornamental， edible and medicinal plant in China， it has many aliases， but it has been recorded in this materia medica under the name of Juhua， Ganju and Ganjuhua. Before the Tang dynasty， medicinal Chrysanthemi Flos mainly collected wild products， including yellow flowers and white flowers， of which the mainstream of yellow flowers was originally Dendranthema lavandulifolium and D. potentilloides， the mainstream of white flowers is D. vestitum and D. chanetii. The cultivation of medicinal Chrysanthemi Flos began in the Northern Song dynasty， and wild D. lavandulifolium， D. potentilloides， D. vestitum and D. chanetii were selected through long-term interspecies and intraspecies crossbreeding， which gradually formed the current cultivar D. morifolium. After chrysanthemums were introduced abroad， foreign scholars began to name chrysanthemums with Linnaeus's plant classification system. In 1792， Mathier named chrysanthemums as Chrysanthemum morifolium and continued to this day， and all the editions of Chinese Pharmacopoeia adopted this scientific name. In the Song dynasty， many local varieties such as Nanyangju， Dengzhouhuang and Dengzhoubai appeared. By the time of the Republic of China， five famous authentic varieties， namely Huaiju， Boju， Chuju， Gongju and Hangju， had been cultivated for medicinal purposes. Boju has been the best medicinal variety since the late Qing dynasty. Hangbaiju has been famous for its tea use， especially the best quality of Huju. Chuju has its own unique characteristics， and it is of good quality both for medicine and tea. Gongju has always been a good tea chrysanthemum. Chrysanthemums are traditionally harvested in September of the lunar calendar， but some of the new varieties cultivated nowadays are harvested earlier. The embryo chrysanthemum in Hangbaiju is a commodity type that collects unopened buds in advance. In ancient times， chrysanthemums were mainly dried in the shade， in modern times， drying methods include drying in the shade， drying in the oven and drying in the sun after steaming. At present， hot air drying is mostly used. In terms of processing， Chrysanthemi Flos was used raw products in ancient times， in modern times， it is still widely used， sometimes stir fried（including stir-fried charcoal）. Due to different varieties， producing areas and processing methods， there are certain differences in the proportion of ingredients contained in chrysanthemum. Therefore， it is suggested that chrysanthemums with different varieties， origins and processing methods should be selected according to clinical indications in the development of famous classical formula preparations containing Chrysanthemi Flos.

Geju Hugan Tablets Ameliorate Alcohol-induced Liver Injury in Mice by Regulating NF-κB and Bcl-2/Bax Signaling Pathways / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Geju Hugan tablets on the liver of mice with alcohol-induced liver injury， and explore the underlying mechanism based on nuclear factor-κB p65 （NF-κB p65） and B-cell lymphoma-2 （Bcl-2）/Bcl-2-associated X protein （Bax） signaling pathways. MethodAccording to the body weight， 60 SPF-grade male ICR mice were randomized into normal， model， Compound Yiganling tablets （0.16 g·kg-1）， and low-， medium-， and high-dose （0.2， 0.4， 0.8 g·kg-1， respectively） Geju Hugan tablets groups. The drugs were administrated at the corresponding doses by gavage， and the normal and model groups with equal volume of pure water once a day for 28 consecutive days. On day 29， the mice in other groups except the normal group were administrated with liquor （53% Vol） by gavage twice a day at the doses of 20， 10 mL·kg-1 and with the interval of 6 h. Samples were harvested on day 30. The histopathological changes in the liver were observed by hematoxylin-eosin （HE） staining， and the ultrastructural changes in hepatocytes were observed by transmission electron microscopy. The enzyme-linked immunosorbent assay was employed to measure the levels of malonaldehyde （MDA）， reduced glutathione （GSH）， and triglycerides （TG） in the liver tissue and the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in the serum. Western blotting was employed to determine the protein levels of NF-κB p65， phosphorylated p-inhibitor kappa B alpha （p-IκBα）， Bcl-2， and Bax in the liver tissue. ResultCompared with the normal group， the model group showed increases in the ALT， AST， MDA， and TG levels， a decrease in the GSH level， and increases in the liver injury scores evaluated based on the HE， oil red O， and transmission electron microscopy （P<0.01）. Moreover， the model group showed up-regulated expression of NF-κB， p-IκBα， and Bax （P<0.05， P<0.01） and down-regulated expression of Bcl-2 （P<0.05） in the liver tissue. Compared with the model group， Geju Hugan tablets of all the doses lowered the ALT， AST， MDA， and TG levels and elevated the GSH level （P<0.01）. The liver injury scores assessed based on HE staining and transmission electron microscopy in the medium- and high-dose Geju Hugan tablets groups were lower than those in the model group （P<0.01）. Compared with the model group， medium- and high-dose Geju Hugan tablets down-regulated the protein levels of NF-κB， p-IκBα， and Bax （P<0.01） and all doses of Geju Hugan tablets up-regulated the protein level of Bcl-2 （P<0.01）. ConclusionGeju Hugan tablets protect mice from alcohol-induced liver injury by down-regulating NF-κB signaling pathway to alleviate inflammation in the liver tissue and down-regulating the expression of Bax and up-regulating the expression of Bcl-2 to inhibit hepatocyte apoptosis.

[Evaluation and optimization of content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia].

This study discovered that the resolution of 3,5-O-dicaffeoylquinic acid(isochlorogenic acid A) in the content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia(ChP)(2020 edition) was poor, which affected accurate quantification. We tested the method in ChP with chromatographic columns of seven brands to clarify the problems in the existing method, optimized the chromatographic conditions by adjusting the mobile phase composition and elution ratio and replacing the chromatographic column packing, and carried out the reproducibility assay for the new method. The two methods were compared for the content determination results of Chrysanthemi Flos prepared from six different varieties. As evaluated by the resolution based on different chromatographic columns of seven brands, the existing method failed to separate isochlorogenic acid A and isochlorogenic acid D well. The peaks of the two components were not completely separated on three chromatographic columns, and isochlorogenic acid A and isochlorogenic acid D generated a co-effluent peak in the other four columns. Isochlorogenic acid A and isochlorogenic acid D could be completely separated under the optimized chromatographic conditions. The difference in the peak areas of isochlorogenic acid A+isochlorogenic acid D obtained by the optimized method and the method in ChP was not significant, with deviation less than 3.0%, which further proved that the result measured by the method in ChP was the co-effluent of isochlorogenic acid A and isochlorogenic acid D. The optimized method can ensure the accurate quantification of isochlorogenic acid A. The existing content determination method of Chrysanthemi Flos has the problem of poor resolution. It is recommended to revise the chromatographic conditions for the content determination method of Chrysanthemi Flos to improve the resolution of isochlorogenic acid A and ensure its accurate quantification.

Evaluation and optimization of content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia / 中国中药杂志

This study discovered that the resolution of 3,5-O-dicaffeoylquinic acid(isochlorogenic acid A) in the content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia(ChP)(2020 edition) was poor, which affected accurate quantification. We tested the method in ChP with chromatographic columns of seven brands to clarify the problems in the existing method, optimized the chromatographic conditions by adjusting the mobile phase composition and elution ratio and replacing the chromatographic column packing, and carried out the reproducibility assay for the new method. The two methods were compared for the content determination results of Chrysanthemi Flos prepared from six different varieties. As evaluated by the resolution based on different chromatographic columns of seven brands, the existing method failed to separate isochlorogenic acid A and isochlorogenic acid D well. The peaks of the two components were not completely separated on three chromatographic columns, and isochlorogenic acid A and isochlorogenic acid D generated a co-effluent peak in the other four columns. Isochlorogenic acid A and isochlorogenic acid D could be completely separated under the optimized chromatographic conditions. The difference in the peak areas of isochlorogenic acid A+isochlorogenic acid D obtained by the optimized method and the method in ChP was not significant, with deviation less than 3.0%, which further proved that the result measured by the method in ChP was the co-effluent of isochlorogenic acid A and isochlorogenic acid D. The optimized method can ensure the accurate quantification of isochlorogenic acid A. The existing content determination method of Chrysanthemi Flos has the problem of poor resolution. It is recommended to revise the chromatographic conditions for the content determination method of Chrysanthemi Flos to improve the resolution of isochlorogenic acid A and ensure its accurate quantification.

Analysis of Chemical Constitutions of Volatile Oil in Opisthopappus taihangensis by GC-MS Combined with Retention Index / 中国实验方剂学杂志

ObjectiveTo study the composition of volatile oil in Opisthopappus taihangensis （Taihangju）， and provide a reference for comprehensive development of this medicine. MethodTaking Chrysanthemum morifolium （Xiaobaiju）， C. morifolium （Xiaohuangju） and C. indicum （Yejuhua） as control， the qualitative and quantitative analysis of volatile oil in Taihangju and three control varieties were completed by gas chromatography-mass spectrometry （GC-MS） combined with retention index method. The GC conditions were as following：programmed temperature （initial temperature at 60 ℃， kept for 2 min； up to 120 ℃ with the heating rate of 5 ℃·min-1， still kept for 2 min； up to 180 ℃ with the heating rate of 2 ℃·min-1， kept for 3 min； and then up to 240 ℃ with the heating rate of 8 ℃·min-1， kept for 5 min； finally up to 280 ℃ with the heating rate of 10 ℃·min-1， kept it for 5 min and finished）， high-purity helium as the carrier gas， the split ratio of 50∶1. MS conditions were as follows：electron impact ion source （EI）， ion source temperature of 230 ℃， electron collision energy of 70 eV and scanning range of m/z 30-445. Principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to obtain the characteristic components between Taihangju and the three control varieties. ResultA total of 86， 96， 112 and 109 compounds including 73 common components were identified in Taihangju， Xiaobaiju， Xiaohuangju and Yejuhua， respectively. The contents of volatile components in Taihangju were significantly different from that of the control varieties. In which， the relative contents of α-thujone， eucalyptol and terpinen-4-ol were high in Taihangju， and eucalyptol， camphor and α-terpinyl acetate were the main compositions in the control varieties. In addition， 11 compounds were screened as characteristic components to distinguish Taihangju and the three control varieties by PCA and OPLS-DA， including main differential components of chamazulene and δ-cadinene. ConclusionThe main components of volatile oil in Taihangju includes alcohols， terpenes， ketones and esters with high medicinal value. The accuracy of qualitative analysis of volatile oil is improved by GC-MS combined with retention index method， which provides scientific reference for development and utilization of Taihangju.

Discovery of Quality Markers of Nucleobases, Nucleosides, Nucleotides and Amino Acids for Chrysanthemi Flos From Different Geographical Origins Using UPLC-MS/MS Combined With Multivariate Statistical Analysis.

Nucleobases, nucleosides, nucleotides and amino acids, as crucial nutrient compositions, play essential roles in determining the flavor, function and quality of Chrysanthemi Flos. The quality of Chrysanthemi Flos from different geographical origins is uneven, but there have been no reports about the screening of their quality markers based on nutritional ingredients. Here, we developed a comprehensive strategy integrating ultra performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UPLC-MS/MS) and multivariate statistical analysis to explore quality markers of Chrysanthemi Flos from different geographical origins and conduct quality evaluation and discrimination of them. Firstly, a fast, sensitive, and reliable UPLC-MS/MS method was established for simultaneous quantification 28 nucleobases, nucleosides, nucleotides and amino acids of Chrysanthemi Flos from nine different regions in China. The results demonstrated that Chrysanthemi Flos from nine different cultivation regions were rich in the above 28 nutritional contents and their contents were obvious different; however, correlation analysis showed that altitude was not the main factor for these differences, which required further investigation. Subsequently, eight crucial quality markers for nine different geographical origins of Chrysanthemi Flos, namely, 2'-deoxyadenosine, guanosine, adenosine 3',5'-cyclic phosphate (cAMP), guanosine 3',5'-cyclic monophosphate (cGMP), arginine, proline, glutamate and tryptophan, were screened for the first time using partial least squares discriminant analysis (PLS-DA) and the plot of variable importance for projection (VIP). Moreover, a hierarchical clustering analysis heat map was employed to intuitively clarify the distribution of eight quality markers in the nine different regions of Chrysanthemi Flos. Finally, based on the contents of selected eight quality markers, support vector machines (SVM) model was established to predict the geographical origins of Chrysanthemi Flos, which yielded excellent prediction performance with an average prediction accuracy of 100%. Taken together, the proposed strategy was suitable to discover the quality markers of Chrysanthemi Flos and could be used to discriminate its geographical origin.

[Changes in chemical compositions of Chrysanthemi Flos after frying and protective effects on CCl_4-induced acute liver injury in mice].

To reveal the processing mechanism of Chrysanthemi Flos from the changes of chemical compositions after frying and its effect on the efficacy of liver protection. Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) and ultra high performance liquid chromatography(HPLC) were used for the qualitative and quantitative researches of chemical compositions before and after Chrysanthemi Flos frying. Progenesis QI and SPSS software were used for principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), variable importance projection(VIP) analysis and t-test to identify the compositions with significant changes. Pharmacodynamics experiment was used to investigate the protective effect of crude and fried Chrysanthemi Flos on CCl_4-induced acute liver injury in mice. According to mass spectrometry data, there were 28 chemical compositions in crude and fried Chrysanthemi Flos, mainly including flavonoids and organic acids. 13 compositions such as luteolin, apigenin and luteolin glycoside were increased significantly after frying, while 7 compositions such as chlorogenic acid, luteolin-7-O-glucuronide and apigenin-7-O-glucuronide were decreased significantly after frying. Through principal component analysis, crude and fried Chrysanthemi Flos products were divided into two categories, indicating that there were internal differences in quality. The results of liver injury protection experiment in mice showed that the AST, ALT and MDA contents were significantly decreased and SOD level was increased in mice with liver injury in both the high and medium dose groups. Histopathological examination showed that crude and fried Chrysanthemi Flos can protect the liver by reducing inflammatory cell infiltration, reducing steatosis, and repairing damaged liver cells. The results of this study showed that the chemical compositions had obvious changes after frying, and both crude and fried Chrysanthemis Flos had protective effects on CCl_4-induced acute liver injury in mice. In addition, in the range of high, medium and low doses, the liver protection effect of crude and fried Chrysanthemi Flos increased with the increase of dose. The experiment results provided reference for the mechanism of fried Chrysanthemi Flos and clinical selection of processed products.

Changes in chemical compositions of Chrysanthemi Flos after frying and protective effects on CCl_4-induced acute liver injury in mice / 中国中药杂志

To reveal the processing mechanism of Chrysanthemi Flos from the changes of chemical compositions after frying and its effect on the efficacy of liver protection. Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) and ultra high performance liquid chromatography(HPLC) were used for the qualitative and quantitative researches of chemical compositions before and after Chrysanthemi Flos frying. Progenesis QI and SPSS software were used for principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), variable importance projection(VIP) analysis and t-test to identify the compositions with significant changes. Pharmacodynamics experiment was used to investigate the protective effect of crude and fried Chrysanthemi Flos on CCl_4-induced acute liver injury in mice. According to mass spectrometry data, there were 28 chemical compositions in crude and fried Chrysanthemi Flos, mainly including flavonoids and organic acids. 13 compositions such as luteolin, apigenin and luteolin glycoside were increased significantly after frying, while 7 compositions such as chlorogenic acid, luteolin-7-O-glucuronide and apigenin-7-O-glucuronide were decreased significantly after frying. Through principal component analysis, crude and fried Chrysanthemi Flos products were divided into two categories, indicating that there were internal differences in quality. The results of liver injury protection experiment in mice showed that the AST, ALT and MDA contents were significantly decreased and SOD level was increased in mice with liver injury in both the high and medium dose groups. Histopathological examination showed that crude and fried Chrysanthemi Flos can protect the liver by reducing inflammatory cell infiltration, reducing steatosis, and repairing damaged liver cells. The results of this study showed that the chemical compositions had obvious changes after frying, and both crude and fried Chrysanthemis Flos had protective effects on CCl_4-induced acute liver injury in mice. In addition, in the range of high, medium and low doses, the liver protection effect of crude and fried Chrysanthemi Flos increased with the increase of dose. The experiment results provided reference for the mechanism of fried Chrysanthemi Flos and clinical selection of processed products.

An integration of UPLC-DAD/ESI-Q-TOF MS, GC-MS, and PCA analysis for quality evaluation and identification of cultivars of Chrysanthemi Flos (Juhua).

BACKGROUND: Chrysanthemi Flos (CF), as a popular traditional Chinese medicine (TCM), has five main cultivars in China, namely "Chuju", "Boju", "Gongju", "Huaiju" and "Hangju". Due to their habitats and processing methods, great quality variations occur yet no systematical study has ever been carried out to evaluate such variations. PURPOSE: In this study, we aim to establish a new approach that can serve both as a quality control method and as an identification method for cultivars of CF. METHOD: The components in CF samples were identified by a combination of UPLC-ESI-Q-TOF/MS and GC/MS. Furthermore, a multimodal quantitative method was established by UPLC-UV coupled with principal component analysis (PCA) and the similarity evaluation system (SES), which was used to control and identify four cultivars of CF. RESULTS: 18 compounds of flavonoids and caffeoylquinic acids were identified and ten of them were quantified using UPLC-ESI-Q-TOF/MS. Different cultivars of CF could be clearly distinguished with the fingerprints evaluation and principal component analysis (PCA). A total of 74 volatile compounds were detected by GC/MS. The distinctness of volatile components was observed. By the combination of UPLC-ESI-Q-TOF/MS and GC/MS, an identification and quality control method for CF was successfully established. CONCLUSION: The combination of UPLC-ESI-Q-TOF/MS and GC/MS could act as a comprehensive multimodal method for both identification and quality control of herbal medicines. This study provided new insights into the overall evaluation method for herbal medicines possessing different cultivars.

B-ring-homo-tonghaosu, isolated from Chrysanthemum morifolium capitulum, acts as a peroxisome proliferator-activated receptor-γ agonist.

The capitula of Chrysanthemum morifolium and C. indicum are used to prepare Chrysanthemi Flos in traditional Japanese Kampo medicine. In our previous study, we reported on the agonistic effect of methanol extract of C. indicum capitulum on peroxisome proliferator-activated receptor (PPAR)-γ. We further isolated (E)-tonghaosu from C. indicum capitulum as one of the active ingredients. In the present study, we aimed to evaluate the PPAR-γ agonistic activity of a methanol extract of C. morifolium capitulum (MCM) in which (E)-tonghaosu could not be detected. MCM exhibited PPAR-γ agonistic activity in a concentration-dependent manner, and at a dose of 100 µg/ml, it showed similar activity to pioglitazone (30 µM), a standard PPAR-γ agonist. Through activity-guided fractionation, we isolated two geometric isomers, (E)- (1) and (Z)-B-ring-homo-tonghaosu (2), as the active ingredients of MCM. Both compounds exerted concentration-dependent PPAR-γ agonistic effects, and 1 had higher activity than 2. At 1.4 µM, 1 had similar activity to pioglitazone (30 µM), which was achieved by 2 at a concentration of 140 µM. Thus, 1 has the potential to become a lead compound for the drug discovery of PPAR-γ agonists. We compared the activities and the contents of (E)-, (Z)-tonghaosu, 1, and 2 among 13 commercial samples of Chrysanthemi Flos, including those derived from both C. morifolium and C. indicum. Their PPAR-γ agonistic activities were not related to the contents of these compounds. 1 and 2 were detected in the samples derived from both species but (E)- and (Z)-tonghaosu were not detected in the samples derived from C. morifolium; hence (E)- and (Z)-tonghaosu can serve as marker compounds to identify the capitula of C. indicum in Chrysanthemi Flos samples.

Imprinting equivalence research of Lonicerae Japonicae Flos and Lonicerae Flos / 中草药

Objective: To explore and confirm the imprinting equivalence of Lonicerae Japonicae Flos (LJF) and Lonicerae Flos (LF) based on the autonomous function of the supramolecular imprinted template, and provide a new solution for double followers dispute. Methods: Using LJF dregs, LF dregs, and Chrysanthemi Flos (CF) dregs as host molecules, and the water extract of LJF, LF, Shuanghuanglian (LJF), Shuanghuanglian (LF), Yinqiaosan (LJF), and Yinqiaosan (LF) as guest molecules, the selective absorb of three host molecules and six guest molecules was carried out. The change of fingerprint of water extract of guest molecules was determinated by HPLC. Then, the MRT and their difference value was calculated through the total quantum statistical moment method,and t-test was performed on it. Results: When the six guest molecules was absorbed by LJF and LF dregs, LJF and CF dregs, LF and CF dregs, the MRT difference value was conducted by t-test. The results were P1 = 0.94 > 0.05, P2 = 0.02 < 0.05, and P3 = 0.04 < 0.05. We can see that all guest molecules was absorbed by LJF and LF dregs. There was no significant difference in the MRT difference value. But when six guest molecules was absorbed by CF dregs. There was significant difference between CF and LJF and CF dreg in the MRT difference value. Conclusion: The statistical data indicated that similarity was existed in both “imprinted template” of LJF and LF. There was difference with CF. It is identified the exist of imprinting equivalence of LJF and LF, which is compatible with clinical medication.

Herbal Textual Research on Genuine Medicinal Materials “Deng Ju” and Quality Evaluation of Nan Yang Chrysanthemi Flos / 中国实验方剂学杂志

Objective: To study the origin of Nanyang Chrysanthemi Flos materia medica and its historical origin, in order to evaluate the quality of Nanyang Chrysanthemi Flos by HPLC method, and define the advantages of Nanyang Chrysanthemi Flos with the origin of Nanyang. Method: Records of Nanyang Chrysanthemi Flos in the "Chinese Medical Code" and related ancient documents were studied to explain the origin and application. The genetic relationship between Nanyang Chrysanthemi Flos and other pieces was revealed in ancient literatures. Then 8 chemical constituents in 14 batches of Chrysanthemi Flos were determined by HPLC multi-components quantitation,and the comprehensive weighted score analysis was performed based on the results. The HPLC fingerprints were established,and the similarity analysis and clustering analysis were made to comprehensive evaluation the quality of Nanyang Chrysanthemi Flos and define the genetic relationship between Nanyang and other pieces at the chemical composition level. Result: The results of the herbal textual research show that Nanyang Chrysanthemi Flos spread from place to place since the Han dynasty,and impact many medicinal chrysanthemums of later generations. HPLC fingerprints similarity and cluster analysis also indicated the genetic relationship between Nanyang Chrysanthemi Flos and other species at the chemical level. The comprehensive score analysis results show that Nanyang Chrysanthemi Flos is of good quality and very suitable for medicinal purposes. Conclusion: Nanyang has been a quality production area of Chrysanthemi Flos since ancient times to present. Nanyang Chrysanthemi Flos is very suitable for medicinal purposes and worthy of promotion.

[Research and application of quality standard for standard decoction of Chrysanthemi Flos].

Standard decoction of traditional Chinese medicine (TCM) is prepared by standardized process, and can be used as references to evaluate the quality of dosage forms such as decoction and dispensing granules. In order to determine the quality evaluation method for standard decoction of Chrysanthemi Flos and investigate its application, 10 batches of white chrysanthemum of Hangzhou were collected to prepare the standard decoction of white chrysanthemum of Hangzhou with standardized process parameters. Parameters such as traits, relative density, pH value, extraction ratio, transfer rate and fingerprint were selected as the indexes for quality evaluation. The established quality evaluation method for standard decoction of Chrysanthemi Flos was applied in the detection of two types of commercial Chrysanthemum dispensing granules. The results showed that the standard decoction of Chrysanthemi Flos was a clear yellow-brown aqueous solution; the relative density was 1.007-1.011; the pH value was between 5.37-5.56; the average extraction ratio was 23.6%, ranging from 19.93% to 29.69%; the average transfer ratewas 56.2% in terms of chlorogenic acid, 57.4% in terms of luteoloside and 30.6% in terms of 3,5-O-dicaffeoylquinic acid. Fingerprint similarity was between 0.864-0.989.The method showed good precision, stability and repeatability in fingerprint analysis, indicating reliable and representative results for standard decoction of Chrysanthemi Flos, and it can be used to evaluate and standardize other dosage forms.

[Construction of high quality Chrysanthemi Flos pieces standard system based on comprehensive analysis of ancient and modern Chinese and foreign pharmacopoeia].

To study the evolution of Chinese ancient and modern pharmacopoeia standards and compare the domestic and foreign pharmacopoeias, further understand the international requirements on chrysanthemum quality, and establish a more suitable and modern standard system for high quality Chrysanthemi Flos pieces. Newly Revised Materia Medica, Welfare Pharmacy, Collected Essentials of Species of Materia Medica (Bencao Pinhui Jingyao), Chinese Pharmacopoeia and other herbal remedies in various generations were reviewed to summarize the evolution of domestic standards on Chrysanthemi Flos pieces. Then they were compared with those in European Pharmacopoeia, United States Pharmacopoeia, Japanese Pharmacopoeia and other foreign Pharmacopoeias to establish a modern and international high-quality Chrysanthemi Flos pieces standard system with Chinese medicine characteristics and produce more internationally recognized high-quality Chinese medicine pieces.

Whole-ingredient identification and primary grade evaluation of Niuhuang Qingwei Pills based on reference drug / 中草药

Objective To identify whole ingredients in Niuhuang Qingwei Pills (NQP) and give primary grade evaluation of NQP. Methods Firstly, microscopic characteristics specified by the statutory standard of NQP were summarized. Then new microscopic identification methods were established for Sennae Folium and Gypsum Fibrosum while new thin layer chromatography (TLC) identification methods were established for Bovis Calculus Artifactus, Borneolum Syntheticum, and Rhei Radix et Rhizoma. In addition, microscopic identification method for Menthae Haplocalycis Herba was improved. In this way, whole-ingredient identification of 17 materials were realized. NQP reference drug was developed and applied as accompanying control in whole-ingredient identification and primary grade evaluation of the samples. Results According to the results of 48 samples from 18 manufactures by primary grade evaluation, 10, 37, and 1 samples were classified as superior, qualified, and unqualified, respectively. Conclusion The proposed methods are accurate, simple, and objective, which offers a more comprehensive approach for quality control of NQP. And the methods provide research strategy and experimental basis for further work on establishment of grade standard.

Research and application of quality standard for standard decoction of Chrysanthemi Flos / 中国中药杂志

Standard decoction of traditional Chinese medicine (TCM) is prepared by standardized process, and can be used as references to evaluate the quality of dosage forms such as decoction and dispensing granules. In order to determine the quality evaluation method for standard decoction of Chrysanthemi Flos and investigate its application, 10 batches of white chrysanthemum of Hangzhou were collected to prepare the standard decoction of white chrysanthemum of Hangzhou with standardized process parameters. Parameters such as traits, relative density, pH value, extraction ratio, transfer rate and fingerprint were selected as the indexes for quality evaluation. The established quality evaluation method for standard decoction of Chrysanthemi Flos was applied in the detection of two types of commercial Chrysanthemum dispensing granules. The results showed that the standard decoction of Chrysanthemi Flos was a clear yellow-brown aqueous solution; the relative density was 1.007-1.011; the pH value was between 5.37-5.56; the average extraction ratio was 23.6%, ranging from 19.93% to 29.69%; the average transfer ratewas 56.2% in terms of chlorogenic acid, 57.4% in terms of luteoloside and 30.6% in terms of 3,5--dicaffeoylquinic acid. Fingerprint similarity was between 0.864-0.989.The method showed good precision, stability and repeatability in fingerprint analysis, indicating reliable and representative results for standard decoction of Chrysanthemi Flos, and it can be used to evaluate and standardize other dosage forms.

Construction of high quality Chrysanthemi Flos pieces standard system based on comprehensive analysis of ancient and modern Chinese and foreign pharmacopoeia / 中国中药杂志

To study the evolution of Chinese ancient and modern pharmacopoeia standards and compare the domestic and foreign pharmacopoeias, further understand the international requirements on chrysanthemum quality, and establish a more suitable and modern standard system for high quality Chrysanthemi Flos pieces. Newly Revised Materia Medica, Welfare Pharmacy, Collected Essentials of Species of Materia Medica (Bencao Pinhui Jingyao), Chinese Pharmacopoeia and other herbal remedies in various generations were reviewed to summarize the evolution of domestic standards on Chrysanthemi Flos pieces. Then they were compared with those in European Pharmacopoeia, United States Pharmacopoeia, Japanese Pharmacopoeia and other foreign Pharmacopoeias to establish a modern and international high-quality Chrysanthemi Flos pieces standard system with Chinese medicine characteristics and produce more internationally recognized high-quality Chinese medicine pieces.