| In traditional Chinese medicine (TCM), the flower bud of Tussilago farfara L., called "Farfarae Flos," also known as "Kuandonghua" in China, has been commonly used for the treatment of cough, bronchitis and asthmatic disorders. Quite many studies of T. farfara L. have been conducting recent years. These studies were mainly on chemical components, pharmacology, toxicity. Cultivation and producing researches also exsit. However, if the traditional use "flower bud of T. farfara L collected during December has higher quality" is rational; if non-traditional use parts have antitussive and expectorant effects; and what the active chemical parts are. These questions have not been answered using modern technology.The bigger, violet, non-rachis flower bud is considered higher quality in the traditional use and the flower bud is used as medicine clinically. According to the theory of TCM, the flower buds of T. farfara are collected in the winter or spring before they are fully open, as the medicinal quality becomes low after blooming and can’t be used as herbal drugs in TCM prescriptions. T. farfara is one of the genuine traditional Chinese medicine in Shanxi province. It is necessary to demonstrate the metabolite content and pharmacological differences during Tussilago farfara L. development, metabolic and pharmacological differences between medicinal parts and non-medicinal parts.In this study, different medicinal parts were used as experimental materials and comparison of chemical components was conducted using NMR-based metabolomics. Antitussive and expectorant effects of different parts (leaf, bud, root, rachis) were evaluated by ammonia induced mice coughing model and volume of phenol red in mice’s tracheas; multi-dimensional analytical methods were used to correlate the chemistry and pharmacology. Finally, based on the former results, further metabolic studies on the different developmental stages, years and sources were conducted.The main results were as follows:(1) Metabolic fingerprinting of different parts (bud, leaf, root) from Tussilago farfara L. The plant materials were collected from Yu county, Hebei province. The metabolic differences were evaluated using NMR-based metabolomics. Through multivariate analysis, we can get two conclusions. A, The scatter plot clear showed that the leaves were separated from the flower buds by PC1. The corresponding loading plot indicated that higher levels of chlorogenic acid, rutin, isoleucine, leucine, valine, threonine, alanine, acetic acid, proline, glutamic acid, aspartic acid, maleic acid fumaric acid, and saturated fatty acids, as well as lower levels of sucrose,(3-glucose, phosphatidyl-choline, choline, malic acid,7β-(3-Ethyl-ciscrotonoyloxy)-1a-(2-methylbutyryloxy)-3(14)-dehydro-Z-notonipetranone, tussilagone, bauer-7-ene-3β,16a-diol, β-sitosterol, and sitosterone were present in leaves as compared with flowers. B, The metabolic variation between the flower buds and roots was also investigated by PCA analysis, which revealed that the root was characterized by more valine, proline, aspartic acid, succinic acid, acetic acid, choline, creatine, glucose, steroids and less aromatic compounds such as caffeic acid, chlorogenic acid,3,5-dicaffeoylquinic acid, rutin and sesquiterpenoids as compared with the flower buds.(2) Metabolic fingerprinting of bud and rachis from Tussilago farfara L Due to the increasing demands of Flos Farfara, adulteration with rachis is frequently encountered in the marketplace. However, to our knowledge, there is no report on the pharmacological and chemical differences between flower bud and rachis. A metabolic profiling carried out by~1H NMR spectroscopy and multivariate data analysis was applied to crude extracts from flower bud and rachis. Principal component analysis (PCA) yielded good separation between flower bud and rachis, and the corresponding loading plot revealed that the rachis was characterized by high signals for the compounds of succinic acid, malic acid, aspartic acid, sucrose,β-sitosterol, sitosterone, fatty acids. The flower bud showed higher aromatic compounds such as caffeic acid, chlorogenic acid,3,5-dicaffeoylquinic acid, rutin and tussilagone. Some amino acids, including threonine, alanine, proline, and glutamic acid, as well as glucose, choline, phosphatidylcholine, acetic acid, formic acid, were also found to be higher in flower bud. Some unidentified signals were also found to largely contribute to the discrimination between flower bud and rachis. These signals mostly belong to sugars and are difficult to identify.(3) Evaluating the antitussive and expectorant properties and correlation between pharmacology and metabolomics of different parts (bud, leaf, root, rachis) from T. farfara L. Ammonia induced mice coughing and phenol red in mice’s tracheas models were established in this study. It was observed that leaf and flower bud groups had significant antitussive and expectorant effects. However, root group almost had no effect, and rachis group had no significant effect on the antitussive expectorant activity. Chlorogenic acid,3,5-dicaffeoylquinic acid, and rutin may be closely related with the antitussive and expectorant activities.(4) Metabolic fingerprinting of different developmental stages of T. farfara L. we collected five different stages from Yu county, Hebei province and conducted the metabolic evaluation. The multivariate analysis revealed that these five different stages had different metabolic fingerprinting. The result of this study was in accordance with the traditional use of this herbal medicine. Metabolites of the early and flowering stage were quite different from those of the traditional harvest time, suggesting that they could not be used as traditional medicine. In addition, antitussive and expectorant effects were the result of synergistic effect of a variety of ingredients.(5) We collected21samples of T. farfara L. from different sources and years to investigate the homogeneous and evaluate the quality using NMR-based metabolomics. Multivariate analysis indicated that these21 samples had different metabolic fingerprinting and this revealed that the quality of T. far far a L. would be different according to the habitats and collecting years.To sum up, our study systematically demonstrated the chemical and pharmacology differences among the different parts and the correlated active chemicals. This research also studied the bud consitituents during five development stages and revealed the chemical differences among samples from different sources. The results have significant meaning in understanding the traditional Chinese medicine theory and broadening our research thinking of T.farfara L. |
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