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Objective: The contents of five main components in Polygala tenuifolia from different habitats were determined, which provided certain data support and theoretical basis for the quality evaluation system of P. tenuifolia. Methods: The contents of polygalaxanthone III, 3,6’-disinapoyl sucrose, polygalacic acid, senegenin and tenuifolin of roots from different wild samples were determined by HPLC. Then SPSS 20.0 and SIMCA 11.5 were used for difference analysis, correlation analysis, hierarchical cluster analysis (HCA), and principal component analysis (PCA). Results: The content of the five main components in wild P. tenuifolia samples from different habitats was significantly different. The 20 samples were placed into two clusters (I, II) by HCA and PCA. Cluster I comprised three samples with higher content of 3,6’-disinapoyl sucrose, polygalaxanthone III, senegenin and tenuifolin from Weinan, Xianyang in Shannxi Province, and Xinjiang in Shanxi Province, whereas cluster II contained the other 17 samples. Conclusion: The results showed that the main components of P. enuifolia from Weinan, Xianyang in Shannxi Province and Xinjiang in Shanxi Province were significantly higher than other origins, and which provided a reference for the quality control, selection of excellent germplasm and cultivation bases of P. tenuifolia.
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Objective: To study the effects of salicylic acid (SA) and methyl jasmonate (MeJA) on growth, activity of related enzymes and chemical components in Polygala callus. Methods: The callus of Polygala was used as material. After 30 d of dark culture at different concentrations of SA (0, 4, 8, 12, 16, 20, 24, 28, 32 mg/L) and MeJA (0, 100, 200, 400, 600, 800, 1 000 μmol/L), the growth of callus, anti-oxidant enzyme activity, the content of MDA, total phenolic, total flavonoid, polygalaxanthone III, and 3,6’-disinapoyl sucrose were determined. Results: MeJA inhibited the growth of Polygala callus, and 12 mg/L SA promoted the growth of Polygala callus. SA and MeJA promoted the activity of SOD, CAT, POD, and MDA in the callus of Polygala. With the increase of SA and MeJA concentration, the activities of SOD, CAT and POD increased first and then decreased, and the content of MDA continued to rise. When the concentration of SA was 20 mg/L, the activities of CAT and SOD reached the maximum, which were 248.45 U/mg and 4451.06 U/mg, respectively. When the concentration of SA was 16 mg/L, the activity of POD reached the maximum, which was 7.22 U/mg. When the concentration of SA was 32 mg/L, the MDA content reached the maximum value of 25.09 nmol/mg. When the concentration of MeJA was 600 μmol/L, the activities of CAT, SOD, and POD reached the maximum, which were 273.30, 1451.06 and 15.27 U/mg, respectively. When the concentration of MeJA was 1000 μmol/L, the MDA content reached the maximum value of 27.10 nmol/mg. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenols. MeJA promoted the accumulation of total phenols and total flavonoids in Polygala callus. When MeJA concentration was 600 μmol/L, the content of total flavonoids was the highest. When the concentration of MeJA was 400 μmol/L, the total phenolic content was the highest. Both SA and MeJA promoted the accumulation of polygalaxanthone III and 3,6’-disinapoyl sucrose in Polygala callus. When the concentration of SA was 32 mg/L, the concentration of polygalaxanthone III, 3,6’-disinapoyl sucrose was the highest. When the concentration of MeJA was 1 000 μmol/L, the content of polygalaxanthone III, 3,6’-disinapoyl sucrose was the highest. Conclusion: MeJA had an inhibitory effect on the growth of Polygala callus, and 12 mg/L SA can promote this effect. SA and MeJA promoted the activity of SOD, POD, CAT, the content of MDA and the accumulation of polygalaxanthone III and 3,6’-disinapoyl sucrose in Polygala callus. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenolics. MeJA promoted the accumulation of total phenolics and total flavonoids in Polygala callus.
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Objective To investigate the effects of different drying methods on composition and content of five active constituents in root bark and root of Polgala tenuifoliaroot. Methods The contents of polygalaxanthone III, 3,6’-disinapoyl sucrose, polygalacic acid, senegenin, and tenuifolin in root bark and root from different drying samples were determined by HPLC. Then the data analysis was performed by ANOVA and TOPSIS methods. Results There is a difference in the order of drying methods for bark and root of P. tenuifoliaroot. For P. tenuifoliaroot root bark, the order of different drying methods was microwave drying > 60 ℃ hot-air drying > 50 ℃ hot-air drying > 70 ℃ hot-air drying > freeze-drying > 40 ℃ hot-air drying > shade drying > sun drying; For P. tenuifolia root, the different drying methods were sorted by microwave drying > 60 ℃ hot-air drying > shade drying > sun drying > 50 ℃ hot-air drying > 40 ℃ hot-air drying > 70 ℃ hot-air drying > freeze-drying. Conclusion Combined with the production practice, this study suggests that microwave drying and hot-air drying at 60 ℃ are suitable drying methods for P. tenuifoliaroot bark and root, providing a basis for the determination of drying methods for the origin processing of P. tenuifolia.
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Objective To study and establish the HPLC fingerprint of Polgala tenuifolia for the identification and quality controlof P. tenuifolia. Methods Twenty batches of wild and cultivated P. tenuifolia collected from different regions in China were detected by HPLC. The Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012 A edition) was used to evaluate the similarity of the samples. The differences among the samples were identified by chemical pattern recognition methods including principal component analysis (PCA) and partial least squares discriminate analysis (PLS-DA). Results The common model of HPLC fingerprint of wild and cultivated P. tenuifolia was obtained, 24 common peaks were found in the chromatograph. The similarities between wild and cultivated P. tenuifolia fingerprints and control fingerprints in 20 batches from different regions were over 0.86, PCA results demonstrated obvious distinction between the wild and cultivated P. tenuifolia. The wild and cultivated P. tenuifolia was completely distinguished by PLS-DA. Twelve constituents, such as polygalactoneone III and 3,6’-disinapoyl sucrose were screened as biomarkers, representing the major differences between the two varieties. Conclusion The HPLC fingerprint combined with chemical pattern recognition can be used as an effective method for the quality control and identification of the different sources of P. tenuifolia, and provide a reference for the quality control and stoichiometric taxonomy of P. tenuifolia.
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Objective To establish the main component analysis method of commodity grade Polygala tenuifolia, and to explore the correlation between the grade character of P. tenuifolia and the main chemistry. Methods P. tenuifolia of four markets were determined by high performance liquid chromatography (HPLC) to analyze the different level of medicine and explore the correlation between commodity grade and composition for P. tenuifolia. Results The main components determination of P. tenuifolia in different market grades were not completely consistent with the grade of commodity. Conclusion There are certain limitations of the commodity grading standards for P. tenuifolia in the medicinal materials market. It is necessary to establish a new grade quality standard for accurately evaluating the quality of P. tenuifolia. This study also provides some reference for the comprehensive quantitative evaluation of P. tenuifolia.
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Objective: A rapid and specific ultra performance liquid chromatography (UPLC) method was established for simultaneous analysis on six compounds and fingerprint analysis on Polygalae Radix to evaluate the herb quality from different habitats in China. Methods: The UPLC method was carried out by gradient elution with acetonitrile-formic acid water (0.1%). The flow rate was 0.4 mL/min. The detection wavelength was at 320 nm. The fingerprint chromatograms and the contents of six compounds including sibiricose A5, sibiricose A6, sibiricaxanthone B, glomeratose A, polygalaxanthone III, and 3,6'-disinapoyl sucrose in 24 batches of Polygalae Radix were analyzed. The common peaks were identified by ultra performance liquid chromatography tandem with time-of-flight mass spectrometry with MSE data-acquistion mode (UPLC-Q-TOF-MSE). Results: There was a difference in contents of six compounds, especially for the content of 3,6'-disinapoyl sucrose and sibiricose A6. Thirty-seven peaks were selected as the common peaks, of which 33 peaks were identified, and the similarities of 24 batches were between 0.756 and 0.997. Based on the results of quantification and fingerprint analysis, a certain difference between samples from different habitats was further proven. Conclusion: The validated UPLC quantitative analysis and fingerprint methods are successfully used in the quality control of Polygalae Radix.
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OBJECTIVE: To carry out a comparison between the cortexes and the roots of Polygala tenuifolia, and to supply references for developing scientific processing method of P. tenuifolia. METHODS: A comprehensive comparison was made by comparing the chemical constitution and pharmacological activities of the cortexes and the roots of P. tenuifolia, i.e., multi-components determination and fingerprint analysis were used to analyze their chemical constituents, and the traditional pharmacological actions of P. tenuifolia, such as cough-relieving, sputum-removing and sedative effects were used to evaluate their efficacy. RESULTS: The chemical constitution of the cortexes, roots and heartwoods of P. tenuifolia was similar, but the contents of the various effective components in the cortexes were more than those in the roots and heartwoods. The pharmacological result revealed that there was no obvious difference between the cortexes and the roots of P. tenuifolia for their activities of cough-relieving, sputum-removing and sedative actions. The roots of P. tenuifolia showed a better tendency than the cortexes for cough-relieving and sputum-removing activities. CONCLUSION: In order to avoid the waste of crude drugs, and to save the cost during heartwood-discarding, the heartwoods of P. tenuifolia are suggested to be kept.
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Occupational asthma is induced by many agents, including herbal materials, that are exposed in working places. Although there are a few case reports for occupational allergy induced by herbal materials, there is none for that induced by Wonji (Polygala tenuifolia). This study was conducted to evaluate clinical characteristics and immunologic mechanism of Wonji-induced asthma in a exposed-worker. A patient who complained of asthma and rhinitis symptoms, and who had worked in a herbal manufacturing factory for 8 yr, underwent a skin prick test with crude extract of Wonji under the impression of occupational asthma induced by the agent. The patient had a strong positive response to the extract on the skin prick test. Allergen bronchial challenge to the extract demonstrated a typical dual response. Serum specific IgE level to the extract was higher in the patient than in healthy controls, and ELISA inhibition test revealed complete inhibition of IgE binding with the extract, but no inhibition with Der p 2 or mugwort extracts. Six IgE binding components to the extract (10, 25, 28, 36, 50, and 90 kDa) were detected using SDS-PAGE and immunoblot analysis. These findings suggest that Polygala tenuifolia, a herbal material, can induce IgE-mediated bronchoconstriction in exposed workers.