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Objective To find the most active phloroglucinol compound on influenza virus from Dryopteris crassirhizoma. Methods Twenty-five phloroglucinols from D. crassirhizoma were docked into 12 kinds neuraminidase (NA) for molecular docking. The binding energies, the combination of compound and neuraminidase and the interaction with the important amino acid residues were used for virtual screening. The inhibitory effect in vitro of four candidate compounds on NA was tested by NA inhibition assay, on this basis, M22 was evaluated for its antiviral activity on MDCK cells via CPE assay. Results Virtual screening suggested that five candidates had strong binding abilities with NA. NA inhibition assay showed that M22 have the strongest activity against NA of all candidates. CPE assays demonstrated that M22 exhibited inhibitory activity on various influenza virus. Conclusion M22 from D. crassirhizoma have significant inhibitory efficacy on influenza virus. The results provide us with useful information for the development of novel anti-influenza drugs.
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OBJECTIVE: To establish an HPLC-MS/MS method for the inspection of illegal use of Woodwardia unigemmata, an adulterant of Dryopteris crassirhizoma, in Kanggan granules. METHODS: The content of kaempferitrin in Woodwardia unigemmata is significantly higher than that in Dryopteris crassirhizoma. Therefore, the content of kaempferitrin can reflect the material usage(Woodwardia unigemmata or Dryopteris crassirhizoma) in Kanggan granules. C18 column was applied with a mobile phase consisting of 0.02% formic acid acetonitrile(V/V)-0.02% formic acid H2O(V/V) in gradient elution mode. The mass spectral analysis was performed in a positive electrospray ionization mode, and scanned by a multiple reaction monitoring mode. The precursor/product ion pair used for quantification was m/z 579.2→287.1. RESULTS: A good linear relationship was obtained in the range of 272.46-5 449.25 pg for kaempferitrin, with the linear regression equation of Y=139.217 2X-2 901.93(r=0.999 6); the average recovery was 94.97%(RSD=0.85%, n=6). Among 12 batches of Kanggan granules, 7 batches from 2 enterprises showed abnormally high contents of kaempferitrin, which indicated illegal use of Woodwardia unigemmata. CONCLUSION: The method is simple, accurate, and reliable, which can be applied to detect the use of Woodwardia unigemmata in Kanggan granules.
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Objective To isolate the phloroglucinol derivatives in Dryopteris crassirhizoma and to discuss its antibacterial activity. Methods The phloroglucinol derivatives were isolated and purified by column chromatography on silica gel, Sephadex LH-20, semi preparative liquid phase, and recrystallization. Structures were proved by physicochemical properties and spectral methods (MS, 1H-NMR, 13C-NMR). The antibacterial activities against fungi and bacteria were tested by CLSI M38-A2 and M07-A9. Results 10 derivatives were obtained from ethanol extracts of D. crassirhizoma, and were identified as 1-butyrylphloroglucinol (1), 1-methyl-3-butyrylphloroglucinol (2), 2-acetyl-4-butyrylphloroglucinol (3), 1-methyl-3-acetyl-5-butyrylphloroglucinol (4), flavaspidic acid PB (5), norflavaspidic acid AB (6), flavaspidic acid AA (7), filixic acid ABA (8), flavaspidic acid AB (9), and filixic acid ABP (10). The antibacterial activity of compound 8 against Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis) was similar to cefoxitin (positive control), and the MIC of compound 8 was 2.5 μg/mL. Conclusion Compounds 1-4, new compounds, are isolated from D. crassirhizoma. Compound 8 has both antibacterial activities against fungi and bacteria.
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To identify origin of the medicinal materials Dryopteridis Crassirhizomatis Rhizoma by using the psbA-trnH sequence, the polymerase chain reaction (PCR) amplification and product sequencing of the experimental samples were performed. In order to expand the scope of the study, the psbA-trnH sequences of 8 genera and 3 species were downloaded from GenBank for analysis. DNAMAN 8.0 software was used to show splicing and comparison results of the peak diagrams with analysis of them, and MEGA 6.0 software was to calculate K2P genetic distances and establish clustering tree adjacent genus. The results showed that by using the psbA-trnH sequence, Dryopteridis Crassirhizomatis Rhizoma, its original plant and other easy-confused medicinal materials and plants can be distinguished with each other obviously, with the psbA-trnH sequence of Dryopteridis Crassirhizomatis Rhizoma completely consistent with that of its original plant. Consequently, it is revealed that it's feasible to identify Dryopteridis Crassirhizomatis Rhizoma and its original plant, and separate from its adulterants by means of the psbA-trnH sequence, which can provide more scientific bases for the further study of the identification of the ferny medicinal herbs.
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Objective: To optimize the separation and purification of the total poly phenol from Dryopteris crassirhizoma with anion- exchange resin. Methods: The rates of elution and absorption, contents of dryocrassin ABBA, and total ploy phenol were used as makers to optimize the purification conditions of total ploy phenol. Results: The extracting solution of D. crassirhizoma passed through 201×7 hydrogen-oxygen the anion-exchange resin column (column diameter-column height, 1:7) at the rate of 6 BV/h in reverse direction, then the resin column was flushed with water at the rate of 6BV/h to pH value 6-7 in forward direction. At last, the column was eluted at the rate of 6 BV/h to obtain ploy phenol with 9BV 3% salt water in 60% alcohol. The elution ratio of dryocrassin ABBA is 90.1%, and the total ploy phenol is 91.1%. The content of dryocrassin ABBA is 29.4% and the content of the total ploy phenol is 49.2%. Conclusion: The separation and purification of the total ploy phenol from D. crassirhizoma with 201 × 7 hydrogen-oxygen anion-exchange resin can achieve the satisfactory results which have wide application prospects.