ABSTRACT
Objective: To establish the HPLC fingerprints of Diphylleia sinensis from different habitats and determine the content of 12 chemical ingredients of picropodophillotoxin-4-O-β-D-glucopyranosy-(1→6)-β-D-glucopyranoside, picropod ophyllotoxin 4-O-glucoside, 4-demethylpodophyllotoxin, podophyllotoxin-4-O-β-D-glucoside, kaempferol-3-O-β-D-glucoside, podophyllotoxin, arabeline, podophyllotoxone, diphyllin-4-O-β-D-glucoside, quercetin, kaempferol, and diphyllin for providing a scientific basis for the quality control of D. sinensis. Methods: COSMOSIL-C18 column (250 mm × 4.6 mm, 5 μm) was used for gradients elution with MeOH (A) -0.4% phosphoric acid solution (B) as mobile phase. Working conditions were as follows: the column temperature was 30 ℃, the flow rate was 1 mL/min, the detection wavelength was 300 nm, and the injection volume was 10 μL. HPLC fingerprint of D. sinensis was established and 12 components were determined. The results were analyzed by cluster analysis. Results: The HPLC fingerprint with 16 common peaks of D. sinensis was established, and the similarities of samples were over 0.9. The linear relationship of 12 components was good (r2≥0.999 0), RSD of precision and repeatability was less than 2%, and the stability was also good with in 24 h (RSD<2%). The average recoveries (n = 6) of 12 components were between 99.27% and 100.3%, and the RSD were in the range of 1.03%-1.98%. The results of the content determination and cluster analysis of twelve components showed that D. sinensis in different habitats were different from each other. Conclusion: This method was simple, sensitive and accurate, which provided a comprehensive reference for the identification and quality evaluation of D. sinensis.
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Objective: To investigate the lignans of the roots of Diphylleia sinensis. Methods: The chemical components were isolated and purified by chromatographic techniques (silica gel, sephadex LH-20 and semi preparative-HPLC) and the chemical structures were determined by spectral data analysis (1D/2D NMR, MS, and IR). Results: Seven lignans were obtained and identified as diphyllin A (1), (1S,2R,5S,6R)-2-(4-hydroxyphenyl)-6-(3-methoxy-4-hydroxyphenyl)-3,7-dioxabicyclo [3.3.0] octane (2), (7S,8R,7’S,8’R)-3,4,3’,4’-tetramethoxy-9,7-dihydroxy-8.8’,7.O.9’-lignan (3), (-)-lariciresinol (4), vladinol D (5), podophyllotoxone (6), and justicidin A (7). Conclusion: Diphyllin A (1) is a new compound and 2-4 are obtained from D. sinensis for the first time.
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Objective: To study the chemical constituents from the radix and rhizome of Diphylleia sinensis. Methods: Silica gel column chromatography, Sephadex LH-20, high performance liquid chromatography, and other chromatographic techniques were used for separation and purification. The structures were elucidated by physical and chemical methods and spectral data. Results: Three compounds were isolated from the 95% ethanol extract in the rhizome of D. sinensis, and their structures identified as diphyllin-4-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (1), justicdin A (2), and (+)-tanegool-7'-methylether (3). Conclusion: Compound 1 is a new compound named procumbenoside G; Compounds 2 and 3 were isolated from the Diphylleia for the first time.
ABSTRACT
Objective To explore a new identification method for medicinal materials of Dysosma, and analyze the second internal transcribed spacer (ITS2) barcode sequences of Diphylleia sinensis and Dysosma versipellis, Sinopodophyllum hexandrum, Dysosma difformis and Dysosma pleiantha in five kinds of podophyllum. Methods The ITS2 of ribosomal DNA of medicinal materials of podophyllum was amplified and sequenced by bi-directional sequencing of PCR products. Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner. Phylogenetic study was performed using software MEGA 5.1 in accordance with Kimura-2-parameter (K2P) model. Genetic distances were calculated and analyzed and the phylogenetic tree was constructed by using the neighbor-joining (NJ) method. Results There were significant differences among five kinds of Dysosma. Their maximum intraspecific genetic distance (K2P distance) was far lower than their minimum interspecific genetic distance with the other species. In the cluster dendrogram, all species showed monophyletic. Conclusion ITS2 sequence as DNA barcoding technique can be used to identify Chinese herbal materials of Dysosma.
ABSTRACT
Diphylleia sinensis is the dried rhizome of Diphylleia sinensis H.L.Li., which belongs to the subfamily of podophyl-lum ( berberidaceae ) , which is always recorded in monograph on materia medica in all ages as one of traditional Chinese medicine“Guijiu” herbal resources .Based on the previous researches in our laboratory and the literatures , the research progress in pharmacog-nosy, chemical constituent, endogeny fungus, quality control, pharmacology and the other aspects of Diphylleia sinensis were systemati-cally reviewed for the comprehensive utilization of its resources , and the development prospects of Diphylleia sinensis was also discussed in the paper , which can provide complete references and ideas for the rational utilization and development of Diphylleia sinensis.
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OBJECTIVE: To establish a method to determine quercetin, podophyllotoxin and kaempferol in Diphylleia sinensis Li. METHODS: Diamonsil C18 column was used. Methanol-0.4% glacial acetic acid solution was used as the mobile phaseat a flow rate of 1 mL · min-1. The column temperature was 30°C. The detection wavelength was 290 nm. RESULTS: The liner ranges of quercetin, podophyllotoxin and kaempferol were 0.023-0.287 μg (r=0.999 9), 0.208-2.6 μg (r=0.999 9), and 0.036 1-0.451 μg (r=0.999 9), respectively. The average recoveries were 99.64% (RSD=0.83%), 100.04% (RSD=0.61%), and 99.88% (RSD=1.19%), respectively. CONCLUSION: The method is sensitive, accurate and reproducible. It can be used for the determination of quercetin, podophyllotoxin and kaempferol in Diphylleia sinensis Li.