Extraction，isolation，identification and content determination of four chemical components in Yao medicine Ventilago leiocarpa / 中国药房

OBJECTIVE To extract and isolate the four chemical components of Yao medicine Ventilago leiocarpa， and to conduct identification and content determination for them. METHODS The chemical components of V. leiocarpa were separated and purified by solvent extraction， extraction， silica gel column chromatography and preparative liquid chromatography； then the chemical structures of four isolated compounds were identified based on their spectral data. The contents of four components were determined by high performance liquid chromatography（HPLC）-quantitative analysis of multi-components by single-marker （QAMS） method， with the following chromatographic conditions： chromatographic column was Echway GowonTM C18 （250 mm× 4.6 mm， 5 μm）. The mobile phase was acetonitrile-0.1% phosphoric acid for gradient elution； the detection wavelength was 269 nm， and the column temperature was 25 ℃ . Using emodin as internal reference， the relative correction factors （fi/s） between emodin and the other 3 components were established and used to calculate the content. At the same time， the content of each component was calculated with the external standard method （ESM）， and the differences between these two methods were compared. RESULTS Four compounds were isolated from V. leiocarpa， and they were identified as emodin， frangulin A， pleuropyrone A， emodin-8-O-β-D-glucoside. The result of HPLC-QAMS showed that the fi/s of pleuropyrone A， emodin-8-O-β-D- glucoside and frangulin A were 1.147 2， 0.874 7 and 0.644 4， respectively. The content of these four components was measured as a good linearity （r≥0.999 6）； relative standard deviation （RSD） of precision， stability and reproducibility tests were all lower than 2.00%， and average recoveries were E-mail：dearhuangjianyou@126.com 99.41%-100.46%（RSD≤2.05%）. There was no significant difference between QAMS method and ESM （RSD＜3.00%）. CONCLUSIONS Emodin， frangulin A， pleuropyrone A and emodin- 8-O-β-D-glucoside are isolated from V. leiocarpa； among them， the last three components are all isolated from for the first time. The established HPLC-QAMS method is accurate and reliable for the determination of 4 components in V. leiocarpa， and can used for quality control of V. leiocarpa.

Research on relationship between color and composition changes in processing of rhubarb charcoal based on "external and internal correlation" / 中草药

Objective: To study the relationship between the quantitative color value of the powder and the known component content of rhubarb charcoal, and lay the foundation for the establishment of the rhubarb charcoal processing process control and endpoint judgment based on the color quantitative value. Methods： Rhubarb charcoal samples were prepared at different temperatures and time. Based on the empirical judgment of the rhubarb charcoal processing, the visual analyzer and UV-Vis were used to quantify the color of the pieces and powder of rhubarb charcoal under different processing conditions. At the same time, the HPLC fingerprint method was used to evaluate the dynamic changes of chemical components during the processing of rhubarb charcoal, and the quantitative value of the color of the sample during the processing of rhubarb charcoal was correlated with the characteristic components of the HPLC fingerprint using the multivariate statistical method. Results: During the processing of rhubarb charcoal, as the degree of carbonization increased, the apparent color of the sample changed from light yellowish brown to burnt black. There was a high correlation between the lightness value (L*), red-green value (a*) of the sample pieces and powder and the yellow and blue values (b*). The area of the 26 characteristic peaks had varying degrees of correlation with the chromaticity value. Among the 14 known components, five bound anthraquinones (aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, chrysophanol-8-O-β-D- glucoside, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside), and two sennosides (sennoside A, sennoside B) had a linear positive correlation with the chromaticity value. The content of five free anthraquinones (aloe-emodin, rhein, chrysophanol, emodin, and physcion), gallic acid and 5-hydroxymethylfurfural (5-HMF), whose contents increased first and then decreased, showed a quadratic correlation with the chromaticity value. Conclusion: The subjective judgment of rhubarb charcoal in the processing process is consistent with the quantitative color value analysis. The quantitative color value has a clear correlation with the content of 14 active chemical components. It is preliminarily inferred that the color quantitative value can be used as the quality of the rhubarb charcoal processing process control and end-point determination indicators to achieve efficient and rapid identification of the processing quality of rhubarb charcoal, which can provide new ideas for the monitoring and quality control of the rhubarb charcoal processing process.

Simultaneous determination of 11 components in raw and processsed products of Rheum pumilum roots by UPLC-ESI-HRMS / 中草药

Objective: To establish an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-ESI-HRMS) determination method for simultaneous determination of 11 components from raw, wine-broiled, carbon-fried and steamed Rheum pumilum roots. Methods: The chromatographic separation was performed on a KinetexTM C18 column (150 mm × 4.6 mm, 2.6 μm) with a gradient elution of acetonitrile and 0.1% formic acid in water at flow rate 0.3 mL/min, the injection volume was 1 μL and column temperature was 32 ℃. The mass spectrometry was detected using ESI ion source and negative ion mode. Results: Eleven components gallic acid, epicatechin, polydatin, rhaponticin, luteolin, emodin-8-O-β-D-glucoside, aloe-emodin, rhein, emodin, chrysophanol and physcion showed a good linear relationship within a certain concentration range. The precision, repeatability and stability of the method were good for the determination of 11 components. The average recoveries varied between 91.31% and 107.08% and the RSD were between 1.73% and 3.58%. The content of gallic acid, polydatin, emodin-8-O-β-D-glucoside and emodin changed in processsed products of R. pumilum roots. The content of gallic acid and emodin increased significantly in the wine-broiled and carbon-fried process. The content of emodin-8-O-β-D-glucoside in the carbon-fried process was significantly reduced, and the content of polydatin was significantly reduced in all processed products. Conclusion: This determination method is simple, stable, accurate and reliable. It can be applied for rapid quantitative determination of 11 components in raw and processsed products of R. pumilum, which laid the foundation for further research on R. pumilum roots.

Quality evaluation of rhubarb based on UPLC-PDA fingerprint and multi-component content determination by chemical pattern recognition / 中草药

Objective: A method was established to obtain fingerprint and determination of ninecomponents in rhubarb based on ultra-performance liquid chromatography photodiode array detection (UPLC-PDA), and 17 batches of rhubarb from different regions, different varieties and different growth years were analyzed. Methods: A ThermoSyncronis C18 column (100 mm × 2.1 mm, 1.7μm) was used with a gradient of acetonitrile (A)-0.1% formic acid (B) as a mobile phase. Fingerprint data was imported intoSIMCA-P 14.1 software for cluster analysis and principal component analysis. At the same time, a total of ninecomponents including sennoside B, rhein-8-O-β-D-glucoside, sennoside A, emodin-8-O-β-D-glucoside, aloe-emodin, rhein, emodin, chrysophanol, physcion were quantitatively analyzed. Results: 20 common peaks were found in the fingerprints of 17 batches of rhubarb, and 9 peaks were identified by standard compounds. Cluster analysis and principal component analysis showed that Rheum tanguticumwas similar to Rheum officinaleand could be distinguished with Rheum palmatumwell. Fouryears and fiveyears of R. tanguticum could not be distinguished, oneyear and twoyears of R.palmatumcould not be distinguished neither. The determination of the indicator components showed thatR. tanguticum was higher than the other two kinds of rhubarb; Fouryears of R. tanguticum was better than five years, and twoyears of R.palmatumwas better than oneyear. Conclusion: This method established rhubarb fingerprint combined with multi-component determination based on UPLC-PDA technology could quickly, scientifically and accurately distinguish rhubarb of different origins. The preliminary evaluation of the rhubarb in different years and a basis for distinguishing the source of rhubarb was also provided.

Preliminary study on hepatotoxic components in Polygoni Multiflori Radix based on different adjuvants processing for detoxification / 中草药

Objective: To provide references for optimizing adjuvants with Polygoni Multiflori Radix (PMR), we compared the effects of detoxification by different adjuvants processing according to Chinese medicine’s records of past dynasties. Methods: The chemical information of all samples including crude and processed PMR with different adjuvants was characterized by UPLC-Q-TOF-MS, and the normal human hepatocytes (L02 cell line) was cultured in vitro to evaluate the cytotoxicity, then we gave synthetic analyses on effects of processed PMR with different adjuvants for toxicity-decreasing and variations of chemical contents. The difference of toxicity reducing effect and the rule of composition change of PMR processed with different adjuvants were compared comprehensively. Results: Different adjuvants had different level of effects on chemical fingerprint, index component and cytotoxicity of PMR under the same conditions of pressure and time. More specifically, black bean, jujube and rice-rinsing water had greater impact on PMR main components including gallic acid, catechins, cis-stilbene glycoside, trans-stilbene glycoside, emodin-8-O-β-D-glucoside, physcion and emodin as well as hepatotoxicity. The three adjuvants with the best toxicity-decreasing effects were in sequence of rice-rinsing water > jujube > black bean. Furthermore, comprehensive analysis of simple correlation and multiple correlation suggested that cis-stilbene glycoside might be the main chemical component contributed to hepatotoxicity of PMR, and emodin-8-O-β-D-glucoside might be the potential toxicity component. Conclusion: Different adjuvants traditionally recorded can attenuate the toxicity of PMR. In addition to black beans, rice-rinsing water and jujube can also be used as candidate adjuvants for the toxicity-decreasing of PMR.

Study on in vitro and in vivo Genotoxicity of Emodin- 8-O-β-D-glucoside / 中国药房

ABSTRACT OBJECTIVE：To evaluate the in vitro and in vivo genotoxicity of emodin-8-O-β-D-glucoside（EG），and to compare the difference of in vitro cell test and in vivo test of rats. METHODS：2D and 3D hepatocyte models were established by in vitro two-dimensional（2D）and three-dimensional（3D）cell culture. After modeling，2D and 3D hepatocyte were divided into blank control group（0.5％ DMSO），mitomycin C group（positive control，0.1 μg/mL），EG low-dose，medium-dose and high-dose groups（10，50，200 μg/mL），respectively. The micronucleus ratio and tail DNA％ of HepaRG cells were detected. SD rats were divided into blank control group（0.5％ sodium carboxymethyl cellulose），ethyl methanesulfonate group（positive control，200 mg/kg），EG low-dose，medium-dose and high-dose groups（100，300，1 000 mg/kg），with 6 rats in each group. They were given medicine intragastrically for consecutive 15 d，once a day. 15 days later，the micronucleus formation rate of bone marrow polychromatic erythrocytes and hepatocytes，the tail DNA％ and tail distance of peripheral blood lymphocytes and hepatocytes were measured. RESULTS：In the in vitro 2D HepaRG hepatocyte model，compared with blank control group，the micronucleus formation rate and tail DNA％ of HepaRG cell were increased significantly in mitomycin C group （P＜0.01）. There was no statistical significance in micronucleus formation rate and tail DNA％ of HepaRG cell among EG groups（P＞0.05）. In 3D HepaRG cell model， compared with blank control group， micronucleus formation rate and tail DNA％ of HepaRG cell were increased significantly in mitomycin C group （P＜0.01 or P＜0.001）， while tail DNA％ of HepaRG cell wasincreased significantly in EG high-dose group（P＜0.01）. In the in vivo test，compared with blank control group，the micronucleus formation rate of bone marrow polychromatic erythrocytes and hepatocytes，the tail DNA％ and tail distance of peripheral blood lymphocytes and hepatocytes were all increased significantly in ethyl methanesulfonate group（P＜0.01）. Tail DNA％ of peripheral blood lymphocytes was increased significantly in EG high-dose group （P＜0.01）. There was no statistical significance in the micronucleus formation rate of bone marrow polychromatic erythrocytes and hepatocytes，the tail DNA％ and tail distance of hepatocytes among EG groups（P＞0.05）；with the increase of dose，there was an increasing trend. CONCLUSIONS：The results of this study suggest that in 2D cell model，EG not lead to chromosome breakage and DNA damage，but the long-term administration and repeated administration in vivo of 3D cell model show that EG has a certain risk of DNA damage，so the evaluation results of 3D HepaRG cell model are more similar to those of rats in vivo. KEYWORDS Emodin-8-O-β-D-glucoside；Genotoxicity；Two-dimensional culture；Three-dimensional culture；Rat；Micronucleus test

Chemical constituents from Rhamnus heterophylla / 中草药

Objective: To investigate the chemical constituents from Rhamnus heterophylla. Methods: Multiple chromatographic separation techniques including silica gel, Sephadex LH-20 gel and MCI gel were employed to isolate and purify the compounds. Their structures were identified by means of the nuclear magnetic resonance (NMR) and physicochemical properties. The chemical compositions were quickly analyzed by HPLC-DAD-ESI-MS/MS and scanned in negative ions. Results: These compounds were isolated and determined as cis-aloeemodin bianthrone (1), trans-aloeemodin bianthrone (2), emodin-8-O-rhamnoside (3), citreorosein (4), emodin (5), chrysophanol (6), physcion (7), kaempferol (8), luteolin (9), and quercetin (10). In addition to compounds 5, 8, the other compounds are isolated from R. heterophylla for the first time, and the compound 3 is isolated from the genus Rhamnus for the first time. 28 Compounds were speculated via comparing MS data (detected mass and MS2 fragment ions) and coupled with related literature, including three hydroxybenzoic acids, eight anthraquinones and 17 flavonoids. Conclusion: The analysis of chemical constituents from R. heterophylla could provide the references for the study on pharmacodynamic material basis and reasonable application and development.

Simultaneous determination of six components from Rumex chalepensis with different growth years and harvesting periods by HPLC / 中草药

Objective: To establish a method for the determination of six components from Rumex chalepensis Mill.. Methods: The contents of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcion were simultaneously determined by HPLC. The mobile phase was methanol-0.1% formic acid, gradient elution, flow rate of 1 mL/min, column temperature of 25 ℃, injection volume of 5 μL, detected by Agilent Extend-C18 (250 mm × 4.6 mm, 5 μm) and diode array detector at 254 nm wavelength. Results: The content of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcionhad good linear relationship in the ranges of 208—3 120, 22.40—336.35, 178.9—2 908.8, 16.7—250.8, 104.4—1 566.0, 45.2—677.7 ng, respectively. The average recovery rates were 97.66%, 97.10%, 98.78%, 97.38%, 102.48%, and 95.51% (n = 6). The contents of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcion in 16 batches of R. chalepensis. were determined in the range of 0.6—7.1, 0—2.7, 1.0—6.5, 0.1—0.6, 0.7—4.3, and 0.1—0.4 mg/g, respectively. Sample contents of different growing years, harvesting dates, and plots were compared and analyzed. Two-year-old R. chalepensis. was collected in early spring or late summer and early autumn. The total content of six components was 12.2 mg/g, which was relatively high. Conclusion: The established method can be used for simultaneous determination of six components from R. chalepensis, and determine the harvesting time and season of R. chalepensis, which provides a scientific basis for the formulation of quality evaluation criteria of R. chalepensis.

Anti-inflammatory Active Ingredient of Chinese Herb Polygoni Cuspidati Rhizoma et Radix Based on Spectrum-effect Relationship / 中国实验方剂学杂志

Objective: To study the potential anti-inflammatory active ingredient of Chinese herbal Polygoni Cuspidati Rhizoma et Radix. Method: The inflammatory model of peritoneal macrophages (RAW264.7 cells) induced by lipopolysaccharide (LPS) in mice was used to screen out the anti-inflammatory activity of 95% ethanol extract of Polygoni Cuspidati Rhizoma et Radix and its macroporous resin elution site (30%, 60%, 95% ethanol eluting site). The characteristic fingerprints were established by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technology, and then partial least squares method (partial least squares, PLS) was used to study the spectrum-effect relationship between the peak area of the characteristic components and the inhibition rate of nitric oxide (NO), and potential anti-inflammatory active ingredients were identified according to variable important in projection (VIP). Result: The ethanol extract macroporous resin 60% ethanol elution site of Polygoni Cuspidati Rhizoma et Radix had the strongest inhibition ability of nitric oxide formation, with a certain dose-dependent relationship. The study of spectrum-effect relationship showed that 3 components had potential anti-inflammatory activity, namely Emodin-8-O-β-D-glucoside (E-8-G), Emodin-1-O-β-D-giucoside, and Emodin-8-O-(6'-O-malonyl)-glucoside. In addition, the anti-inflammatory activity of E-8-G was further validated at the cell level through molecular docking analysis. Conclusion: Three potential anti-inflammatory active ingredients were found base on the spectrum-effect relationship. This study strategy is helpful to find the active ingredients (group) of traditional Chinese medicine, and provides new research ideas and methods for studying the material basis of Chinese herbal medicine.

Rheum officinale at different years old stage / 中草药

Objective To analyze the content and variation rules of 10 constituents in radix, rhizome, and leaf of Rheum officinale at one-, two-, and three-year-old stage, respectively, and provide theoretical guidance for efficient production and quality control of the crud drug. Methods The content of each constituent in R. officinale was determined by high performance liquid chromatography (HPLC), and one factor analysis of variance and multiple comparison were performed by SPSS 24.0. Results HPLC system was established for the determination of 10 components in R. officinale. The linear range was good (r2 > 0.997), RSD of precision, stability, and repeatability were less than 2%, and the recoveries were 96.10%—107.10%, respectively. The content analyses showed that, in the same part, the content of gallic acid decreased significantly year by year or at the 2nd growth years (P 3 > 2 (P 1 > 2 (P radix > leaf (P < 0.05). Conclusion The HPLC based determination of 10 constituents in R. officinale showed that the accumulation profiles of the samples at different years or from different parts varied. For the same parts, the contents of most constituents increased year by year. During the same growth year, the contents of most constituents in radix or rhizome were higher than those in leaf. The radix and rhizome of the three years old samples had the highest contents of main constituents.

Chemical components from Lianhua Qingwen Capsules (II) / 中草药

Objective: To study the chemical constituents of Lianhua Qingwen Capsules. Methods: The compounds were isolated and purified by gel column chromatography, MPLC, and preparative HPLC from 50% ethanol fraction of macroporous resin column chromatography of Lianhua Qingwen crude extracts. Their structures were elucidated by the spectral analyses. Results: Eight compounds were isolated and identified as 10-O-(p-hydroxycinnamoyl)-adoxosidic acid (1), aloe-emodin-8-O-β-D-glucopyranoside (2), quercitrin (3), matairesinol-4’-O-β-D-glucoside (4), liquiritin apioside (5), epi-vogeloside (6), vogeloside (7), and caffeic acid ethyl ester (8). Conclusion: Compound 1 is a new compound named lianhua iridoid A. Compounds 5-8 are isolated from Lianhua Qingwen Capsules for the first time. This study provides substance foundation for chemical research of Lianhua Qingwen Capsules.

Effects of aqueous extract of Polygoni Multiflori Radix and its main constituents on expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells / 中草药

Objective To analyze the main chemical constituents and their contents in aqueous extract of Polygoni Multiflori Radix (PMR, root of Polygonum multiflorum), and to elucidate the effects of aqueous extract of PMR and its main constituents on the expression of the mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells. Methods The main chemical constituents and their content in aqueous extract of PMR were determined by HPLC. The cytotoxicity of aqueous extract of PMR and its main constituents on L02 cells was determined by MTT assay. The mRNA expression of CYP1A2, CYP2C9, and CYP2E1 in L02 cells were detected by quantitative real-time PCR. Results There were four main well-separated chromatographic peaks standing for tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside, emodin and physcion in aqueous extract of PMR. The content of thesecomponents in aqueous extract of PMR was (1.14 ± 0.03)%, (0.106 9 ± 0.001 6)%, (0.010 8 ± 0.000 9)%, (0.003 55 ± 0.000 19)%, respectively. The cytotoxicity of aqueous extract of PMR and emodin on L02 cells at 24 h was dose-dependent, and the concentration of 50% inhibition was 7.290 mg/mL and 0.082 mmol/L respectively. Tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside and physcion did not show significant cytotoxicity on L02 cells in the experimental concentrations. Aqueous extract of PMR and emodin significantly inhibited the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells. Emodin-8-O-β-D-glucoside inhibited the expression of mRNA of CYP1A2 and CYP2C9. Tetrahydroxy stilbene glucoside inhibited the expression of mRNA of CYP1A2 but activated the expression of mRNA of CYP2C9. Physcion inhibited the expression of mRNA of CYP1A2 and CYP2C9 in a dose-dependent manner, but inhibited the expression of mRNA of CYP2E1 in low concentration and activitated the expression of mRNA of CYP2E1 in high concentration. Conclusion The inhibition of aqueous extract of PMR on the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells is the combined effect of all components in it. The main four components all inhibit the expression of mRNA of CYP1A2. The anthraquinone is the main component inhibiting the expression of mRNA of CYP2C9. The free anthraquinone is the main component inhibiting the expression of mRNA of CYP2E1.

Comparision of contents of thirteen components in different origins of Rhei Radix et Rhizoma based on principal component analysis / 中草药

Objective To establish an HPLC method for the simultaneous determination of thirteen effective components in Rhei Radix et Rhizoma and to assess the quality from different producing areas. Methods The analysis was performed on ZORBAX C18 chromatographic column with ographic mobile phase consisted of methanol-0.1% phosphoric acidwater in a gradient mode. The flow rate was 1.0 mL/min. Gallic acid, catechin, epicatechin, aloe-emodin-8-O-glucopyranoside, rhein-8-O-glucopyranoside, emodin-8-O-glucopyranoside, physcionin-8-O-glucopyranoside, chrysophanol-8-O-glucopyranoside, aloe-emodin, rhein, emodin, chrysophanol and physcionemod were determined at 254 nm, and the results were analyzed by principal component analysis (PCA). Results The linear regression equation of 13 components of Rhei Radix et Rhizoma was established within a certain concentration range. The precision and reproducibility were less than 5.00%, the recovery of the method was 95.54%-103.19%, and the sample was stable within 24 h. The contents of Rhei Radix et Rhizoma in 27 batches were differences. Five principal components were obtained by PCA, the principal component integrated model was between 0.335 and 1.905 and the value of Rhei Radix et Rhizoma in Mianyang of Sichuan Province was the highest. Conclusion A method for the determination of thirteen main components of Rhei Radix et Rhizoma was established. PCA was used to evaluate the quality of Rhei Radix et Rhizoma and the main component model value can be used as the evaluation index of Rhei Radix et Rhizoma.

Preliminary study on hepatotoxic components in Polygoni Multiflori Radix based on processing and toxicity-decreasing / 中草药

Objective: To explore the main toxic components induced hepatotoxicity based on the spectrum-toxicity correlation analysis of processed Polygoni Multiflori Radix (PMR), and to provide reference of PMR for promoting the quality control as well as the safe clinical treatment. Methods: The UPLC-Q-TOF-MS characterized chemical information of all samples including crude and processed PMR which were black soybean steamed with high pleasure in different time, and the main components were identified after referring to the literatures. With the normal human hepatocytes (Lo2 cell line) as in vitro model and cell inhibitory rate as testing index, the simple correlation analysis and multivariate linear correlation analysis were used to screen the main components of PMR hepatotoxicity. Result: Seven main same components of crude and processed PMR were identified as trans-stilbene glycoside, gallic acid, emodin, physcion, emodin-8-O-β-D-glucoside, cis-stilbene glycoside, and catechin. And it was found that trans-stilbene glycoside, physcion, emodin-8-O-β-D-glucoside, cis-stilbene glycoside, and catechin were closely related to PMR hepatotoxicity. Furthermore, physcion and cis-stilbene glycoside were found greatly contributing to hepatotoxicity induced by PMR in the principle component regression analysis, which indicated these two might be the main toxic components. In addition, the results demonstrated apparent effects in detoxification could not be achieved until black soybean steamed with high pleasure at least 36 h. Conclusion: This study will provide data support for further rational use of PMR and deep hepatotoxicity research.

HPLC method for simultaneous determination of nine components from leaves of Rheum officinale by HPLC / 中草药

Objective: To establish an HPLC method for determing nine components from the leaves of Rheum officinale, such as gallic acid, senna glycosides B, rhubarb phenol-1-O-glucoside, emodin-8-O-glucoside, aloe-emodin, rhein, emodin, rhubarb, and emodin methyl ether, as well as to explore the scientific evidence for reasonable exploitation and utilization of medicinal rhubarb leaf. Methods: Takeing C18 chromatographic column (250 mm × 4.6 mm, 5 microns), methanol-0.2% acetic acid water as mobile phase, gradient elution and flow rate of 1 mL/min, column temperature 30℃, and detection wavelength of 260 nm. Results: Gallic acid, senna glycosides B, rhubarb phenol-1-O-glucoside, emodin-8-O-glucoside, aloe-emodin, rhein, emodin, rhubarb, and emodin methyl ether had the good linear relationship, respectively in the range of 20.2-606.0, 79.3-2 379.0, 10.1-301.8, 14.8-443.7, 0.7-2.2, 0.13-3.9, 12.4-372.0, 38.8-1 164.0, 6.2-185.4 ng (r > 0.999 6); The nine kinds of the ingredients of the average recovery were 95.76% - 95.76%, RSD was 1.46% - 2.43%. Conclusion: This method is simple and accurate, and with good effect and reliable results for the separation and determination of the components from the leaves of R. officinale, which can provide the reference for its rational development and utilization.

Separation and purification of emodin-8-O-β-D-glucopyranoside from Polygoni Cuspidati Rhizoma et Radix by column and high-speed counter-current chromatography / 中草药

Objective: To develop an effective and rapid method for the preparation of emodin-8-O-β-D-glucopyranoside (EG) reference substance from Cuspidati Rhizoma et Radix (PCRR). Methods: The target fraction was isolated by macroporous resin column chromatography and ODS column chromatography, and then the target compound was purified by high-speed counter-current chromatography (HSCCC). The structure was identified by ESI-MS and NMR spectral techniques, and its purity was determined by HPLC analysis using the main component self-comparison with no correction factor method. Results: A solvent system that consisted of dichloromethane-ethyl acetate-methanol-water (8:1:6:5) was applied to the separation based on assessment using HPLC partition coefficient method. The upper phase was used as the stationary phase, while the lower phase was used as the mobile phase. The reference substance of EG was prepared, and its purity was up to 98% in line with the standard of quantitative analysis. Conclusion: The method is suitable for the preparation of EG from PCRR, which provides a basis for the quality control of natural product and their preparations.

Influence of prescription compatibility on extraction rate of 15 constituents in Yinchenhao Decoction / 中草药

Objective: To research the influence of prescription compatibility on the extraction rate of main constituents in Yinchenhao Decoction (YCHD). Methods: The extraction volume of two iridoids (geniposide and deacetylasperulosidic acid methyl ester), two crocetin derivatives (crocin I and crocin II), three bound anthraquinones (aloe-emodin-8-O-glucoside, chrysophanol-1-glucoside, and emodin-8-glucoside), five free anthraquinones (aloe-emodin, rhein, emodin, chrysophanol, and physcion), two tannins (gallic acid and catechin), and chlorogenic acid in Artemisiae Scopariae Herba (ASH), Gardeniae Fructus (GF), Rhei Radix et Rhizoma (RRR), the compatibility of ASH and GF (compatibility A), the compatibility of ASH and RRR (compatibility B), the compatibility of GF and RRR (compatibility C), and the compatibility of ASH, GF, and RRR (compatibility D) were determined by HPLC. Taking the extraction volume in single medicine as 100%, the extraction rates of 15 constituents mentioned above in compatibilities A-D were calculated. Results: The extraction rates of geniposide, crocin I, and crocin II in compatibilities A, C, and D, deacetylasperulosidic acid methyl ester in compatibility C, gallic acid in compatibilities C and D, and chlorogenic acid in compatibilities A-D decreased. The extraction rates of deacetylasperulosidic acid methyl ester in compatibilities A and D, three bound anthraquinones, aloe-emodin, and chrysophanol in compatibility B, chrysophanol and physcion in compatibilities C and D increased. Conclusion: Prescription compatibility has some influence on the extraction rate of main constituents in YCHD.

Studies on nootropic activity and mechanism of emodin-8-O-?-D-glucopyranoside / 中草药

Object To investigate the nootropic activity of emodin 8 O ? D glucopyranoside, the active principle of Radix Polygoni Multiflori Preparata(PMEG), and its mechanism of action Methods Nootropic activity was evaluated with step down test of mice, and its nootropic mechanism studied by its antiacetylcholinesterase (anti AchE) effect on mice brain in vivo and in vitro as determined by pectrophotometry Results PMEG (ig) significantly improved the learning and memory of both normal and intelectual deficit mice induced by scopolamine PMEG showed dose dependent inhibition of AchE, and the antiacetylcholinesterase effect is reversible both in vivo and in vitro In acute experiment, the half recovery time of AchE is 115 min, and 165 min in chronic experiment Conclusion PMEG showed good nootropic activity, and it is a reversible AchE inhibitor