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AIM To determine effective components of Ampelopsis grossedentata (Hand.-Mazz.) W.T.Wang under different breeding modes and to analyze the antioxidant enzyme activities.METHODS Single factor and orthogonal tests were adopted in screening the optimal extraction process of antioxidant enzymes and analyzing antioxidant enzyme activities of A.grossedentata under common cutting,two-phase cutting and tissue culture.Solvent extraction method was adopted in extracting the effective components of three breeding modes in stem leaves of A.grossedentata,whose contents were determined.RESULTS In three breeding modes,the content of polysaccharides in tissue culture seedling was the highest,while the contents of flavonoids and polyphenols in the two stage cutting seedlings were the highest.The optimal extraction process of antioxidant enzymes was determined to be 6.0 for pH,20 ℃ for processing temperature and 20 min for processing time.Antioxidant enzyme activities were in sequence of two stage cutting seedlings ＞ common cutting seedling ＞ tissue culture seedling.CONCLUSION Considering the contents of effective components and antioxidant enzyme activities,two stage cutting method is the best among 3 breeding modes.
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Objective: To screen reference genes for real time quantitative PCR (qRT-PCR) research in Ampelopsis grossedentata. Methods: On the basis of the conserved sequences among plant species, six candidate reference genes (including Actin, 18 S-rRNA, GAPDH, α-Tubulin, β-Tubulin, and UBQ) were cloned from A. grossedentata by RT-PCR in this study. The expression stability of each reference gene in different tissues (shoot tip, young leaf, mature leaf, old leaf, stem, and root) were analyzed by three softwares (GeNorm, NormFinder, and BestKeeper), followed by validation of the expression pattern of AgPAL by qRT-PCR. Results: Actin, 18 S-rRNA, and GAPDH expressed most stably in all samples and were suitable for reference genes, which were further confirmed by the transcript level analysis result of AgPAL in different tissues. Conclusion: This is the first report on the screening and validation of reference genes for qRT-PCR in A. grossedentata, which benefits future studies on gene expression in this species.
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In recent years, researches on the antibacterial experiments of single herb, extracts, monomer ingredients and the compound preparations have confirmed that a variety of Chinese meteria medica have good antibacterial effects, and they are not easily produced drug resistance. Ampelopsis grossedentata, which can be used as medicine and food, contains various biological ingredients. Among them, flavonoids have desired antibacterial effect and synergistic effect when combined with antibiotics. In this review, antibacterial effects of A. grossedentata were summarized from domestic and foreign study literatures in the recent decade, including the main antibacterial components, the antimicrobial effects, and mechanism of this medicinal plant, aming to provide reference for the studies on the physiological and pharmacological functions, clinical application, and product development of A. grossedentata.
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Objective: A new quality evaluation method using HPLC fingerprint coupled with quantitative analysis of multi-components by single marker method was developed for the quality control of Ampelopsis grossedentata. Methods: HPLC analysis was performed on an Agilent Eclipse Plus C18 (250 mm × 4.6 mm, 5 μm) column using methanol-0.05% aqueous solution of phosphoric acid as mobile phase at flow rate of 0.8 mL/min. The column temperature was set at 50 ℃ and the detection wavelength was at 258, 292, and 369 nm. The quality analyses of samples from different origins were accomplished by cluster analysis and principal component analysis using SPSS 19.0 statistical software. Results: The HPLC fingerprint of A. grossedentata from different regions was established. Twenty-four common peaks were found in the HPLC fingerprint of 14 samples, three important common components were identified as dihydromyricetin, myricitrin and myricetin. Further, the quantitative analysis of multi-components by single marker method of these contents was established. Conclusion: The method of HPLC fingerprint combined with quantitative analysis of multi-components by single marker method can be used for the quality control of A. grossedentata.
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OBJECTIVE: To study the chemical constituents of Ampelopsis grossedentata (Hand-Mazz) W.T. wang. METHODS: The chemical constituents were isolated and purified with silica column chromatography, gel chromatography, etc. Their structures were identified by physicochemical properties and various spectroscopic methods including NMR spectrum, MS, UV, etc. RESULTS: Sixteen compounds were isolated and elucidated as dihydromyricetin(1), physcion(2), quercetin(3), myricetin(4), taxifolin(5), kaempferol(6), dihydrokaempferol(7), myricetin-3-O-L-rhamnoside(8), afzelechin(9), bellidifod-in(10), quercetin-3-O-α-L-rhamnopyranoside(11), myricetin-3'-O-β-D-xylopyranoside(12), ampelopsin(13), myricetin-3-O-β-D-glucoside(14), astragalin(15) and myricetin-3-O-β-D-galactopyranoside(16). CONCLUSION: Compound 10 is reported from the plants in genus Ampelopsis for the first time. Compounds 5,6,7,9,12 and 15 are isolated from this plant material for the first time.
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Object To establish a method for the assay of myricetin in Ampelopsis grossedentata (Hand. Mazz.) W. T. Wang. Methods The determination was carried out with RP HPLC on Nova pak C 18 stainless steel column (150 mm ? 3 9 mm) with methanol∶water (40∶60) as the mobile phase, and detected at a UV wave length of 254 nm. Results The coefficient of variation was 2.763% with average recoveries=97.4%. The minimal detectable limit was 15 ng. Contents of myricetin in different parts of A. grossedentata from Hunan Province varied from 1.57% to 2.17%. Conclusion The method is rapid, simple, accurate and good for the determination of myricetin in A. grossedentata.