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Objective: Clematis florida var. plena is a traditional She medicine. In this paper, we aim to study the main chemical components of essential oil of C. florida var. plena flower. The study will provide reference for large-scale reasonable cultivation and quality standard establishment of C. florida var. plena. Methods: C. florida var. plena flower's essential oil was extracted with steam distillation and it was identified by GC-MS. Area normalization method was used to measure the percentage of each components. Results: In the experiment, 32 element were detected in essential oil of C. florida var. plena flower, among which 20 chemical components were identified. There are 13 chemical components over 1%, and the top five compounds are palmitic acid (26.94%), phytol (10.58%), linoleic acid (6.13%), pentadecane (4.54%) and n-tricosane (3.84%). Conclusion: GC-MS was first used to analyze the main chemical components of C. florida var. plena's flower essential oil. The study provides scientific basis for cultivation, production, development, utilization of C. florida var. plena.
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Objective: To discriminate traditional She medicine Gegongniugen (the root of Rubus chingii Hu) from its confusable species of genus Rubus based on ITS2 barcode, and to guarantee the medicine quality and clinical effect of Gegongniugen. Methods: A total of 140 samples of R. corchorifolius L. f., R. hirsutus Thunb., R. parvifolius L., R. coreanus Miq., R. buergeri Miq., R.tsangorum Hand. -Mazz., R. trianthus Focke, R. jiangxiensis Z. X. Yu, W. T. Ji et H. Zheng were collected from Zhejiang, Guangxi, Jiangxi, and Anhui provinces. Genomic DNA was extracted from the collected samples, followed by ITS2 sequences amplification through PCR, bi-directional sequencing, assembly and annotation based on HMMer method. At the same time, 37 other sequences of corresponding species were downloaded from GenBank. Sequence characteristics such as lengths, GC contents, and variable sites were analyzed by Gene Tool software while genetic distance calculation and build-up of neighbor-joining (NJ) phylogenetic tree were carried out through Clustal X and MEGA 7.0. Results: The lengths of ITS2 sequences of Gegongniugen and its nine confusable species of genus Rubus ranged from 211 to 213 bp with GC content of 52.1%-58.0%; These ITS2 sequences had 14 mutation sites and one insertion/deletion site; The average intra-specific K2P genetic distance of Gegongniugen was 0.007 2, which was significantly lower than the respective inter-specific K2P genetic distance between Gegongniugen and its confusable species of genus Rubus. The genetic distantes between Gegongniugen and R. corchorifolius or Gegongniugen and R. jiangxiensis were minimum, while that between Gegongniugen and R. hirsutus was maxmum. It was obvious in the N-J phylogenetic tree that the ITS2 sequences of Gegongniugen formed into one separate clade, which could be successfully distinguished other nine species. Conclusion: ITS2 sequence can be used as an effective DNA marker to discriminate the original plants of Gegongniugen and its confusable species of genus Rubus, which provided an important molecular evidence for the identification of traditional She medicines and contributed to the information supervision of traditional She medicine market.
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Objective: To establish an assay method of flavonoids (four components) in traditional SHE medicine-Shiliang Tea by quantitative analysis of multi-components by single-marker (QAMS), and to analyze the dynamic change of flavonoids at different harvest time and different collection places. Methods: With rutin as the internal reference substance, the relative correction factor (RCF) of kaempferol-3-O-rutinoside, quercetin, and kaempferol was calculated. Then the contents of four components were calculated, and the accuracy and feasibility of method was evaluated through external standard method. Results: The RCF of rutin to kaempferol-3-O-rutinoside, quercetin, and kaempferol were 1.158 with RSD 0.73%, 0.475 6 with RSD 1.55%, 0.431 9 with RSD 1.58%, respectively. There was no significant difference of assay results between QAMS method and external standard method. While the differences of content between different harvest months and two different species were significant. Conclusion: The QAMS method with rutin as internal reference substance can be used for quantitative analysis of four flavonoids in Shiliang Tea. It is suggested that the best harvest time of Shiliang Tea for flavonoids is in July and August.
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Objective: To explore the chemical constituents of traditional She medicine Rubi Radix et Rhizoma (the roots and rhizomes of Rubus chingii in family Rosaceae). Methods: The compounds were isolated and purified by various column chromatography and the structures were identified by physicochemical constant determination and spectral analysis. Results: Nine compounds were isolated and identified as β-sitosterol (1), daucosterol (2), euscaphic acid (3), 11α-euscaphic acid (4), tormentic acid (5), ellagic acid (6), gallic acid (7), ursolic acid (8), and oleanolic acid (9) from 95% ethanol extract of Rubi Radix et Rhizoma. Conclusion: All the nine compounds are isolated from Rubi Radix et Rhizoma for the first time and the compound 4 is first isolated from R. chingii.
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Objective:To study the quality standard for Herba Desmodii,and provide the scientific basis for a reasonable method of quality control. Methods:With the methods of microscopic identification,thin-layer chromatography and high performance liquid chromatography for the qualitative and quantitative determination of Herba Desmodii,the ethanol soluble extracts,moisture content and total ashes of Herba Desmodii were determined. Results:The characteristics of thin-layer chromatography were obvious;a good lineari-ty of quercetin and kaempferol was within the range of 0. 002-0. 050 μg(r=1. 000 0)and 0. 004-0. 100 μg(r=0. 999 9)with the av-erage recovery of 97.89%(RSD=0.89%,n=6)and 98.93%(RSD=1.31%,n=6),respectively. Conclusion:The method is simple,accurate and reproducible,which can effectively control the quality of Herba Desmodii.
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Objective:To establish an HPLC method for the determination of oleanolic acid in She medicine radix of Aralia chinen-sis L. . Methods:The HPLC analysis was performed on a Waters XBridge-C18 (250 mm × 4. 6 mm, 5 μm) column. The mobile phase was methanol-0. 1 mol·L-1 ammonium acetate solution (83∶17) and the flow rate was 1. 0 ml·min-1 . The detection wavelength was 210 nm, the column temperature was 20 ℃, and the injection volume was 10 μl. Results:Oleanolic acid in radix of Aralia chinensis L. had a good separation from the other components, a good linearity was obtained within the range of 72. 52-725. 2 μg·ml-1 ( r=1. 000 0), and the average recovery was 98. 05%(RSD=1. 89%, n=6). Conclusion:The method is simple, accurate, reproducible and applicable in the assay of oleanolic acid in radix of Aralia chinensis L. .
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Objective: To establish an HPLC method for the simultaneous determination of three flavonoids in Shi Liang Cha. Methods:HPLC was applied with the chromatographic conditions as follows: the chromatographic column was Agilent Zorbax SB-C18 (250 mm × 4. 6 mm, 5μm) at 30℃;acetonitrile-0. 1% H3 PO4 solution was used as the mobile phase with gradient elution; the flow rate was 1. 0 ml·min-1, and the detection wavelength was 360nm. Results: Good linearity of rutin, quercetin and kaempferol was within the range of 0.0409-1.637 0mg·ml-1(r=0.999 2), 0.44-88.00μg·ml-1(r=0.999 8) and 0.41-77.63μg·ml-1(r=0. 999 2), respectively; the average recovery was 99.35%(RSD =1. 64%),101. 14% (RSD =1. 88%) and 99. 69% (RSD =1. 92%) , respectively. Conclusion:The method is simple, accurate and repeatable, and suitable for the simultaneous determination of rutin, quercetin and kaempferol in Shi Liang Cha.