Color characterization and its correlation with quality index during ripening of Rubus chingii / 中国中药杂志

The color of Rubus chingii was characterized by digital method, and the content of water extract, alcohol extract, total flavonoids, total polysaccharides, total polyphenols, ellagic acid, linden glycoside, kaophenol-3-O-rutin were determined. Correlation regression was used to analyze the correlation between color and composition. The results showed that L~* was positively correlated with total polyphenols, kaophenol-3-O-rutin and tilide, and moderately positively correlated with total flavones, ellagic acid and aqueous extracts. The a~* value was negatively correlated with total polyphenols, kaophenol-3-O-rutin, and linden glycosides, while was moderately correlated with total flavones, aqueous extracts, and ellagic acid. The b~* value was negatively correlated with the water extract, and moderately correlated with the content of total polyphenols, total polysaccharides, alcohol extract and kaophenol-3-O-rutin, which showed that R. chingii mature color had a significant correlation with material composition in the process of dynamic change. According to the law of dynamic change in the color and quality indexes, it is determined that the appropriate harvest time is in late April to May 1, while the fruit is not turn yellow. The agronomic traits related to fruit was(12.49±0.56) mm in diameter,(14.25±1.19)mm in height,(1.20±0.14) g in weight, the chroma L~* value was 52.87±3.14,a~* value was 2.01±1.58, b~* values was 28.31±3.88. The results lay a foundation for establishing an objective quantitative evaluation model of R. chingii color from experience.

Effective medicinal ingredients and screening of excellent germplasm in Rubus chingii / 中国中药杂志

In order to provide rationale for selection of good germplasm in Rubus chingii, main effective medicinal ingredients of green fruit such as gallic acid, ellagic acid, kaempferol-3-rutinoside, astragalin and tiliroside were measured using UPLC for the samples collected from Chun'an county of Zhejiang province, and such parameters as soluble solid contents of ripe fruit of some samples were also measured to study variation among individuals and correlation. It has been found that there were differences among individuals in the contents of gallic acid, ellagic acid, kaempferol-3-rutinoside, astragalin and tiliroside, which ranged from 0.010 2%-0.027 4%, 0.089 5%-0.291 1%, 0.010 5%-0.114 8%, 0.005 8%-0.041 2% and 0.010 9%-0.086 3%, respectively, with a CV of 18.60%, 27.02%, 44.23%, 44.17% and 47.29%, respectively. Gallic acid was positively correlated with ellagic acid, but negatively with kaempferol-3-rutinoside and astragalin significantly. Significantly positive correlation existed between kaempferol-3-rutinoside, astragalin and linden glycoside as well as between ellagic acid and fruit shape index of ripe fruit and between linden glycoside and the content of soluble solids. 51.35% of the individuals had a content of soluble solids more than 15%. Therefore, abundant variations have been found among individuals in effective medicinal ingredients in R. chingii, which shows great potential for selection, but only do 7.61% of the individuals meet the requirement of Chinese pharmacopoeia in terms of the contents of effective medicinal ingredients. Therefore, selection could be first performed in terms of fruit shape index of ripe red fruit, followed by the contents of ellagic acid and kaempferol-3-rutinoside measured. The individuals, in which the contents of effective medicinal ingredients don't meet the requirement of Chinese pharmacopoeia, could be considered for the selection in terms of edible fresh fruit.

Rubus chingii Hu: an overview of botany, traditional uses, phytochemistry, and pharmacology / 中国天然药物

Rubus chingii Hu, a member of the rosaceae family, is extensively distributed in China and Japan. Its unripe fruits (Fupenzi in Chinese) have a long history of use as an herbal tonic in traditional Chinese medicine for treating various diseases commonly associated with kidney deficiency, and they are still in use today. Phytochemical investigations on the fruits and leaves of R. chingii indicate the presence of terpenoids, flavonoids, steroids, alkaloids, phenylpropanoids, phenolics, and organic acids. Extracts or active substances from this plant are reported to have various pharmacological properties, including antioxidant, anti-inflammatory, antitumor, antifungal, antithrombotic, antiosteoporotic, hypoglycemic, and central nervous system-regulating effects. This review provides up-to-date information on the botanical characterizations, traditional usages, chemical constituents, pharmacological activities, toxicity, and quality control of R. chingii. Possible directions for future research are also briefly proposed. This review aims to supply fundamental data for the further study of R. chingii and contribute to the development of its clinical use.

Protein tyrosine phosphatase 1B inhibitory activities of ursane-type triterpenes from Chinese raspberry, fruits of Rubus chingii / 中国天然药物

Protein tyrosine phosphatase 1B (PTP1B) has led to an intense interest in developing its inhibitors as anti-diabetes, anti-obesity and anti-cancer agents. The fruits of Rubus chingii (Chinese raspberry) were used as a kind of dietary traditional Chinese medicine. The methanolic extract of R. chingii fruits exhibited significant PTP1B inhibitory activity. Further bioactivity-guided fractionation resulted in the isolation of three PTP1B inhibitory ursane-type triterpenes: ursolic acid (1), 2-oxopomolic acid (2), and 2α, 19α-dihydroxy-3-oxo-urs-12-en-28-oic acid (3). Kinetics analyses revealed that 1 was a non-competitive PTP1B inhibitor, and 2 and 3 were mixed type PTP1B inhibitors. Compounds 1-3 and structurally related triterpenes (4-8) were further analyzed the structure-activity relationship, and were evaluated the inhibitory selectivity against four homologous protein tyrosine phosphatases (TCPTP, VHR, SHP-1 and SHP-2). Molecular docking simulations were also carried out, and the result indicated that 1, 3-acetoxy-urs-12-ene-28-oic acid (5), and pomolic acid-3β-acetate (6) bound at the allosteric site including α3, α6, and α7 helix of PTP1B.

Identification of traditional She medicine Gegongniugen and its confusable species of genus Rubus using ITS2 barcode / 中草药

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.

Simultaneous determination of four flavonoids in Rubus chingii by MIPs-HPLC / 中成药

AIM To establish a molecular imprinted polymers (MIPs)-HPLC method for the simultaneous content dertermination of four flavonoids in Rubus chingii Hu.METHODS Fe3O4 magnetic MIPs was added into ethyl acetate fraction solution of R.chingii to prepare mixed MIPs.The analysis of mixed MIPs methanol eluent was performed on a 30 ℃ thermostatic Diamonsil C18 column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 266 nm.RESULTS Tannins,lilsonide,quercetin and kaempferol showed good linear relationships within the ranges of 2.4-1 232.0 μg (R2 =1)、7.1-3 648.0 μg (R2 =0.999 9)、4.7-4 840.0 μg (R2 =0.999 9)、4.8-2 440.0 μg (R2 =0.999 9),whose average recoveries were 98.94％,99.33％,99.26％ and 98.67％ with the RSDs of 2.04％,1.40％,1.76％ and 1.75％,respectively.CONCLUSION This accurate and reliable method eliminates impurity interference,which can be used for the content determination of flavonoids in R.chingii.

Simultaneous determination of four flavonoids in Rubus chingii by MIPs-HPLC / 中成药

AIM To establish a molecular imprinted polymers (MIPs)-HPLC method for the simultaneous content dertermination of four flavonoids in Rubus chingii Hu.METHODS Fe3O4 magnetic MIPs was added into ethyl acetate fraction solution of R.chingii to prepare mixed MIPs.The analysis of mixed MIPs methanol eluent was performed on a 30 ℃ thermostatic Diamonsil C18 column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 266 nm.RESULTS Tannins,lilsonide,quercetin and kaempferol showed good linear relationships within the ranges of 2.4-1 232.0 μg (R2 =1)、7.1-3 648.0 μg (R2 =0.999 9)、4.7-4 840.0 μg (R2 =0.999 9)、4.8-2 440.0 μg (R2 =0.999 9),whose average recoveries were 98.94％,99.33％,99.26％ and 98.67％ with the RSDs of 2.04％,1.40％,1.76％ and 1.75％,respectively.CONCLUSION This accurate and reliable method eliminates impurity interference,which can be used for the content determination of flavonoids in R.chingii.

Simultaneous Determination of Eight Constituents in Fruits of Rubus chingii by UPLC / 中草药·英文版

Objective To develop a simple, efficient, and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii, which is widely used in Chinese materia medica (CMM), known as Fupenzi (FPZ) in Chinese. Methods An ultra performance liquid chromatography-photo diode array (UPLC-PDA) system was employed for simultaneous quantification of eight compounds, i. e. adenosine, gallic acid, brevifolin carboxylic acid, ethyl gallate, ellagic acid, kaempferol-3-O-rutinoside, kaempferol-3-O-β-D-glucopyranoside, and tiliroside. The chromatographic analysis was performed on a C18 column using a gradient elution of acetonitrile −0.1% formic acid aqueous solution within a runtime of 25 min. Results All calibration curves were linear (R2 > 0.9997) over the tested ranges. The intra- and inter-day precisions as determined from sample solutions were both less than 2.45% and 2.78%, respectively. The average recoveries for the eight constituents ranged from 94.77% to 101.35% with RSD ≤ 4.41%. The newly-developed method was applied to the quality assessment of various R. chingii samples, including both ripe and unripe fruits of R. chingii from different habitats. Conclusion The relative levels of the investigated compounds vary remarkably in the fruits of R. chingii collected from different habitats. As only two of the eight compounds, adenosine and ellagic acid, are determined in the ripe fruits of R. chingii, the results may explain the reason why only the unripe fruits can be used in CMM.

Chemical constituents of traditional She medicine Rubi Radix et Rhizoma / 中草药

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.

Extraction Technology Optimization and Content Determination of Gallic Acid in the Roots of Rubus Chin-gii Hu from Eastern China / 中国药师

Objective:To optimize the extraction technology of gallic acid in the roots of Rubus chingii Hu from eastern China and establish an assay method. Methods:The content of gallic acid was determined by HPLC. Orthogonal design was used to investigate the influence of solid/liquid ratio,extraction time and concentration of hydrochloric acid on the yield of gallic acid. Results:A good linearity of gallic acid was within the range of 4. 082-40. 815 μg·ml-1(r=0. 999 9),and the average recovery was 101. 28%(RSD=1. 08%,n=6). The optimal technology was as follows:extracted with heating circumfluence in 4. 2% hydrochloric acid solution with solid/liquid ratio of 1: 15 for 2. 5h. Conclusion:The optimal extraction technology is simple and feasible,and the assay method is stable and reliable,which can be used in the extraction and determination of gallic acid in the roots of Rubus chingii Hu from eastern China.

Study on chemical constituents of methylene chloride extract of Rubus chingii / 第二军医大学学报

Objective: To study the chemical constituents of the methylene chloride of Rubus chingii Hu.. Methods: Chromatography on silica gel column, Sephadex LH-20 column, and recrystallization technique were used to isolate and purify the compounds. Spectroscopy methods including EI-MS, ES-MS, 1H-NMR, 13CNMR, HMQC, and HMBC were used to elucidate the structures of compounds. Results: Ten compounds were obtained and 9 compounds were identified as: hexacosanol (I), β-sitosterol (II), 4-hydro-3-methlbenzal acid (III), 4-hydrobenzal dehyde (IV), oleanolic acid (V), stigmast-5-en-3-ol, oleate (VI), H-2-indenone, 2, 4, 5, 6, 7, 7a-hexahydro-3-(1-methylethyl)-7a-methyl (VIII), 4-hydroxy-3-methoxybenzoic acide (IX), and liballinol (X). Conclusion: Compounds I, III, IV, VI, VIII, and X have been obtained from Rubus chingii for the first time.

Study on chemical constituents of methylene chloride extract of Rubus chingii / 第二军医大学学报

Objective:To study the chemical constituents of the methylene chloride of Rubus chingii Hu..Methods:Chromatography on silica gel column,Sephadex LH-20 column,and recrystallization technique were used to isolate and purify the compounds.Spectroscopy methods including EI-MS,ES-MS,1H-NMR,13CNMR,HMQC,and HMBC were used to elucidate the structures of compounds.Results:Ten compounds were obtained and 9 compounds were identified as:hexacosanol(Ⅰ),?-sitosterol(Ⅱ),4-hydro-3-methlbenzal acid(Ⅲ),4-hydrobenzal dehyde(Ⅳ),oleanolic acid(Ⅴ),stigmast-5-en-3-ol,oleate(Ⅵ),1H-2-indenone,2,4,5,6,7,7a-hexahydro-3-(1-methylethyl)-7a-methyl(Ⅷ),4-hydroxy-3-methoxybenzoic acide(Ⅸ),and liballinol(Ⅹ).Conclusion:Compounds Ⅰ,Ⅲ,Ⅳ,Ⅵ,Ⅷ,and Ⅹhave been obtained from Rubus chingii for the first time.

HPLC fingerprint of Rubus chingii Hu / 中成药

AIM:To establish HPLC fingerprint for controlling the quality of Rubus chingii Hu. METHODS: Analysis was performed on Alltech C_(18) column(250 mm?4.6 mm,5 ?m)with a mixture of acetonitrile and water as mobile phase in gradient mode,flow rate was 1.0 mL/min,wavelength at 211 nm. RESULTS: There were 15 common peaks in the HPLC fingerprints of Rubus chingii Hu.The RSD of precision and reproducibility lay within 5%. CONCLUSION: The method has good sensitivity and repeatability.This chromatographic fingerprint method can be used to controll the quality of Rubus chingii Hu.