Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells / 中草药·英文版

Objective: Chrysophanol (Chry) displays potent anticancer activity in human cancer cells and animal models, but the cellular targets of Chry have not been fully defined. Herein, we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity. Methods: Human liver cancer cell line HepG2 was incubated. The cytotoxicity was evaluated by MTT assay. Mitochondria localization was evaluated by a confocal microscopy. Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer. The levels of ATP, mitochondrial superoxide anions, and GSH/GSSG were determined according to the assay kits. The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining, respectively. The expression of cyclophilin D (CyPD) was determined by immunoblot method, and the interaction between CyPD and Chry was analyzed by molecule docking procedure. Results: Chry itself mainly localized in mitochondria to cause mitochondrial dysfunction and cell death in HepG2 cells. As regard to the mechanism, cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore (mPTP) moderately suppressed cell death, indicating mPTP involved in the process of cell death. Further, Chry enhanced the protein expression of Cyclophilin D (CyPD) which is a molecular componentry and a modulator of mPTP, while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD. Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with −11.94 kal/mol of the binding affinity value. Besides, the mtDNA-deficient HepG

Effects of Chrysophanol on the Activation of Microglia and the Expression of Inflammatory Factors in Cerebral Ischemia Model Rats / 中国药房

OBJECTIVE：To study the effects of chrysophanol on the activa tion of microglia and the expression of inflammatory factors in cerebral ischemia model rats. METHODS ：SD rats were randomly divided into sham operation group ， model group and chrysophanol high ，medium，low dose groups [7.88，3.94，1.97 mg/（kg·d）]，with 20 rats in each group （the number was complemented in cases of death or unsuccessful modeling during modeling process ）. Except for sham operation group ， middle cerebral artery occlusion model was established in other groups by improved thread method. After 2 hours of ischemia ， sham operation group and model group were intraperitoneally injected with 1 mL normal saline ，and each administration group was intraperitoneally injected with 1 mL corresponding drug ，once a day ，for 7 consecutive days. After last medication ，the score of neurological impairment was recorded ；cerebral infarction of rats was observed by TTC staining ，and the percentage of cerebral infarction area was calculated. The expression of Iba- 1 positive cells in ischemic penumbra of rats was observed by immunofluorescence staining. The expression of Notch- 1，TNF-α and ICAM-1 in the ischemic penumbra of rats were detected by Western blotting assay. RESULTS ：In sham operation group ，there was no infarction area in the brain tissue ，and the Iba- 1 positive cells in the ischemic penumbra were few and branched. Compared with sham operation group ，the infarction area of cerebral tissue in rats was obvious in model group ； the 052）number of Iba- 1 positive cells in ischemic penumbra were 〔ZQ2017003〕） increased significantly ，and they were in amoeba or round shape；the neurological impairment score ，the percentage of cerebral infarction area ， relative expression of Notch- 1， TNF-α and ICAM-1 protein in ischemic penumbra were increased significantly （P＜0.05）. Compared with m odel rats ，the infarction area of cerebral tissue in each dose group of chrysophanol was reduced to different extent ；the number of Iba- 1 positive cells in ischemic penumbra was decreased ；neurological impairment score ，the percentage of cerebral infarction area ，relative expression of Notch- 1，TNF-α and ICAM-1 protein were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Chrysophanol has a certain protective effect on the brain tissue of cerebral ischemia model rats ，and can relieve the nerve injury. Its mechanism may be associated with inhibiting the activation of microglia and expression of inflammatory factors mediated by Notch pathway.

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.

Mechanism of anti-hepatoma of Rhei Radix et Rhizoma based on integrative pharmacology platform, GEO database chip and molecular docking / 中草药

Objective: To investigate the anti-hepatoma active components of Rhei Radix et Rhizoma and their molecular mechanisms through GEO database, integrative pharmacology platform and molecular docking technology. Method: The active ingredients of Rhei Radix et Rhizoma were screened by TCMIP and the corresponding targets of these components were predicted through TCMIP and Swisstarget databases. The hepatoma gene chip database was downloaded from GEO databases, and the differentially expressed genes between hepatocellular carcinoma (HCC) and normal liver tissue were analyzed by GEO2R. Based on the matching results of potential targets of Rhei Radix et Rhizoma and the targets of hepatoma, the key targets of Rhei Radix et Rhizoma against hepatoma were screened, and GO function enrichment and KEGG pathway enrichment analysis of the key targets were performed. Main components and core targets of Rhei Radix et Rhizoma against hepatoma were analyzed and screened by constructing PPI network, component-target network and traditional Chinese medicine-component-target-pathway network. Furthermore, the molecular docking between the core targets and the main active components was performed by Schrodinger-Maestro software to virtually verify their binding ability and analyze their binding mode. Result: A total of 20 anti-hepatoma active components of Rhei Radix et Rhizoma were collected and related 86 targets were obtained, including CDK1, AKR1C3, PTGS2, AR and CCNB1, etc. The results of GO functional enrichment mainly focused on the cell cycle, G2/M transition of mitotic cell cycle, oxidation-reduction process, drug reactions and steroid metabolism processes, etc. The results of KEGG pathway enrichment mainly involved cell cycle, cell senescence, complement system, arachidonic acid metabolism and bile metabolism, and these metabolic pathways were related to cell apoptosis, metastasis, inflammation and immunity. The results of molecular docking showed that 92.2% of the active components had good binding ability with the 10 core proteins, and the main combination forms mainly were hydrogen bonds, hydrophobic bonds, π-π bonds and cation-π. Conclusion: The active components of Rhei Radix et Rhizoma including rhein, emodin, chrysophanol-8-O-β-D-glucopyranoside, chrysophanol-1-O-β-D-glucoside and rhapontigenin can act on multiple targets such as CDK1, CCNB1, CYP2C9, MMP9 and PTGS2, by regulating signaling pathways related to cell apoptosis, metastasis, inflammation and immunity to play an anti-hepatoma effect.

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.

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 Analysis of Four Standards of The Herbal Formula, DF-02, of Ephedra intermedia and Rheum palmatum, using by High Performance Liquid Chromatography-Ultraviolet Detector (HPLC-UVD)

The herbal formula, DF-02, consisting of Ephedra intermedia and Rheum palmatum are used for the treatment of the metabolic diseases such as obesity and liver fibrosis in Korean local clinics. We aimed to develop the simultaneous analytical conditions for four standards, (+)-pseudoephedrine (PSEP) and (−)-ephedrine (EP) for E. intermedia, and aloe-emodin (AE) and chrysophanol (CP) for R. palmatum using HPLC-UV techniques. The validated conditions yielded the high precision (relative standard deviation (RSD) 0.9994). As a result, four standards of DF-02 were simultaneously determined under the developed method, which will be utilized for the quality control or evaluation of DF-02 and many herbal preparations containing E. intermedia and R. palmatum.

Effect of different proportions of piper chinaroot and rhubarb on the dissolution rate of five effective components in rhubarb / 国际中医中药杂志

Objective@#To study the effect of different proportions of piper chinaroot and rheum palmatum on the dissolution rate of five effective components (aloe emodin, emodin acid, emodin, emodin, emodin methyl ether).@*Methods@#The high performance liquid chromatography (HPLC) method was used to analyz the contents of five effective components of rheum palmatum in the extracts of different combination of piper chinaroot and rheum palmatum. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by gradient elution with methanol and 0.1% phosphoric acid water solution as mobile phase at a flow rate of 1ml/min, the column temperature was 30 ℃ and the detection wavelength was 254 nm.@*Results@#The linear ranges of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 0.018 5-0.741 8, 0.017 9-0.717 8, 0.015 9-0.635 5, 0.054 2-2.167 2, 0.016 2-0.646 4 μg, respectively. The average recoveries of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 94.35%, 95.50%, 100.61%, 96.27%, 97.39%, and the RSDs were 1.81%, 1.99%, 2.84%, 2.71%, and 1.86%, respectively. Compared with rheum palmatumby single extract, the content of aloe emodin and emodin were increased by 5 proportions (3:1, 2:1, 1:1, 1:2, 1:3), while the content of emodin and emodin methyl ether were decreased.@*Conclusions@#The optimal compatibility proportion of piper chinaroot and rheum palmatum is 1:1.

Chrysophanol protects against doxorubicin-induced cardiotoxicity by suppressing cellular PARylation

The clinical application of doxorubicin (DOX) in cancer chemotherapy is limited by its life-threatening cardiotoxic effects. Chrysophanol (CHR), an anthraquinone compound isolated from the rhizome of L., is considered to play a broad role in a variety of biological processes. However, the effects of CHR׳s cardioprotection in DOX-induced cardiomyopathy is poorly understood. In this study, we found that the cardiac apoptosis, mitochondrial injury and cellular PARylation levels were significantly increased in H9C2 cells treated by Dox, while these effects were suppressed by CHR. Similar results were observed when PARP1 activity was suppressed by its inhibitors 3-aminobenzamide (3AB) and ABT888. Ectopic expression of PARP1 effectively blocked this CHR׳s cardioprotection against DOX-induced cardiomyocyte injury in H9C2 cells. Furthermore, pre-administration with both CHR and 3AB relieved DOX-induced cardiac apoptosis, mitochondrial impairment and heart dysfunction in Sprague-Dawley rat model. These results revealed that CHR protects against DOX-induced cardiotoxicity by suppressing cellular PARylation and provided critical evidence that PARylation may be a novel target for DOX-induced cardiomyopathy.

Simultaneous determination of nine constituents in Jiuwei Gantai Capsules by HPLC-DAD / 中草药

Objective To establish HPLC-DAD method for the simultaneous determination of allantoin, ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R1, baicalin, schisandrin, curcumin, emodin and chrysophanol in Jiuwei Gantai Capsules (JGC). Methods The chromatographic separation was achieved on an Ultimate AQ-C18 column (150 mm × 4.6 mm, 5.0 μm) with mobile phase consisted of (0.1% phosphate)-(acetonitrile-methanol 20 : 80) for gradient elution, at the flow rate of 1.0 mL/min; The column temperature was 35 ℃. Results The linear ranges of allantoin, ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R1, baicalin, schisandrin, curcumin, emodin, and chrysophanol were 1.6—160.0 μg/mL (r = 0.999 7), 1.2—120.0 μg/mL (r = 0.999 6), 1.2—120.0 μg/mL (r = 0.999 6), 0.4—40.0 μg/mL (r = 0.999 2), 4.0—400.0 μg/mL (r = 0.999 8), 0.4—40.0 μg/mL (r = 0.999 1), 0.16—16.0 μg/mL (r = 0.999 1), 0.08—8.00 μg/mL (r = 0.999 0), and 0.2—20.0 μg/mL (r = 0.999 2), respectively. The average recoveries (n = 6) were 99.3% (RSD = 0.6%), 100.2% (RSD = 0.5%), 99.8% (RSD = 0.7%), 98.3% (RSD = 1.1%), 99.9% (RSD = 0.3%), 97.8% (RSD = 0.9%), 97.8% (RSD = 1.4%), 102.2% (RSD = 1.5%), and 101.9% (RSD = 1.2%), respectively. The content of the allantoin, ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R1, baicalin, schisandrin, curcumin, emodin, and chrysophanol in nine batches were 3.634—3.655, 2.523—2.611, 2.405—2.424, 0.802—0.829, 10.362—10.623, 0.901—0.921, 0.334—0.366, 0.142—0.160, and 0.462—0.479 mg/g, respectively. Conclusion The method is accurate, sensitive, credible, and repeatable, which can be applied to the quality control of JGC.

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.

Simultaneous determination of eleven constituents in Chenxiang Huazhi Pills by HPLC / 中草药

Objective To develop an HPLC method for simultaneous determination of the eleven constituents (agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion) in Chenxiang Huazhi Pills (CHP) by HPLC with gradient elution. Methods The chromatographic separation was performed on an Thermo Syncronis C18 column (4.6 mm × 250 mm, 5 μm) which was operated at 30 ℃. The mobile phase was a linear gradient prepared from water (A) and acetonitrile (B). The linear gradient elution program was programmed as follows: 0—10 min, 20% acetonitrile; 10—20 min, 20%—40% acetonitrile; 20—24 min, 40% acetonitrile; 24—26 min, 40%—52% acetonitrile; 26—30 min, 52% acetonitrile; 30—31 min, 52%—90% acetonitrile; 31—35 min, 90% acetonitrile; 35—40 min, 90%—100% acetonitrile; 40—43 min, 100% acetonitrile; 43—45 min, 100%—20% acetonitrile. The flow rate was 1 mL/min and the detection wavelength was 215 nm. Results The analysis permitted very good separation of eleven constituents within 43 min. A good linear relationship between the peak area and the injection volume was obtained. The ranges of the eleven constituents were 1.4—13.6, 10.0—200.0, 31.5—315.0,1.0—120.1, 1.8—50.6, 0.93—10.1, 1.8—30.0, 0.2—40.3, 1.8—18.1, 1.7—25.0, and 0.45—10.70 μg/mL. The average recoveries of eleven constituents in the samples were in the range of 98.90%—100.87%. The precision RSD of the peak areas of the 11 components ranged from 0.55%—1.54%; Eleven components had good stability within 30 h, and the concentration RSD of each component ranged from 0.75% to 1.94%; The repeatability RSD of each component ranged from 0.39% to 1.73%. The content of agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion in six batches were 92.0—201.0, 511.5—9 033.0, 5 475.0—12 635.5, 54.5—5 095.5, 192.0—2 137.5, 117.0—391.5, 106.5—1 281.5, 13.0—136.5, 93.5—199.0, 177.0—1 207.0, and 33.5—251.5 μg/g, respectively. Conclusion The method is accurate, rapid and simple with high sensitivity, precision and repeatability, which has been successfully applied as an effective tool for the multicomponent analysis of CHP.

Comparison of 17 constituents in rhubarb before and after honeyed wine processing in Yi medicine / 中草药

Objective To compare the changes of 17 constituents in rhubarb before and after the processing of honeyed wine. Methods HPLC method was used to quantitatively analyze 17 components including gallic acid, catechin, epicatechin, polydatin, ferulic acid, sennoside B, rhein-8-O-β-D-glucoside, sennoside A, emodin-1-O-glucoside, chrysophanol-8-O-glucoside, kaempferol, emodin-8-O-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion in honeyed wine rhubarb. Results Seventeen kinds of ingredients, gallic acid, catechin, epicatechin, polydatin, ferulic acid, sennoside B, rhein-8-O-β-D-glucoside, sennoside A, emodin-1-O-glucoside, chrysophanol-8-O-glucoside, kaempferol, emodin-8-O-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion, were well separated. The linear ranges of which were 25.94—830.00 μg/mL (r = 0.999 1), 28.20—1 805.00 μg/mL (r = 0.999 8), 77.03—2 465.00 μg/mL (r = 0.999 2), 25.00—1 600.00 μg/mL (r = 0.999 3), 11.41—730.00 μg/mL (r = 0.999 6), 7.85— 1 005.00 μg/mL (r = 0.999 0), 210.47—6 735.00 μg/mL (r = 0.999 0), 113.28—3 625.00 μg/mL (r = 0.999 6), 10.94—700.00 μg/mL (r = 0.999 8), 218.80—1 415.00 μg/mL (r = 0.999 6), 55.00—1 760.00 μg/mL (r = 0.999 2), 48.44—1 550.00 μg/mL (r = 0.999 7), 19.22—625.00 μg/mL (r = 0.999 6), 18.91—1 210.00 μg/mL (r = 0.999 1), 14.06—450.00 μg/mL (r = 0.999 2), 61.41—1 965.00 μg/mL (r = 0.999 4), and 25.16—805.00 μg/mL (r = 0.999 5), respectively. The averages of the total recovery were 93.71%—102.77% at the three concentrations of 17 components. The content of sennoside and anthraquinone was significantly reduced after rhubarb was processed by Yi method, while the content of gallic acid was significantly increased. Conclusion For the first time, HPLC method was used to quantitatively analyze the 17 components in honeyed wine rhubarb, which provided a reference for the quality standards of honeyed wine rhubarb.

Quality evaluation of Dahuang Lidan Tablets based on multi-component simultaneous quantitative and statistical analysis / 中草药

Objective: A method for simultaneous determination of 10 active ingredients in Dahuang Lidan Tablets (DLP) by HPLC wavelength switching method was established, and its quality was evaluated by statistical analysis. Methods: A Phenomenex Kinetex C18 column was used with a column temperature of 30 ℃ and a mobile phase gradient of methanol-0.15% phosphoric acid. The flow rate was 1 mL/min and the detection wavelengths were 265.0 nm (0-5.8 min, gallic acid), 283.9 nm (5.8-7 min, 5-hydroxymethyl furfural), 222.2 nm (7-18 min, corilagin, p-hydroxybenzaldehyde), 256.7 nm (18-74 min, ellagic acid, aloe-emodin, rhein, emodin, chrysophanol, emodin methyl ether), respectively. Statistical analysis of the content of components in 10 batches of drugs was performed using SPSS 21 Software. Results: The linearity of 10 components in the respective mass concentration ranges was good (r > 0.998 0), the average sample recovery was in the range of 98.45%-100.12%, and the RSD was in the range of 0.80%-2.51%. The content of 10 components was as follow: gallic acid (8.371-11.438 mg/tablet), 5-hydroxymethylfurfural (0.046-0.087 mg/tablet), corilagin (0.721-2.094 mg/tablet), p-hydroxybenzaldehyde (0.034-0.065 mg/tablet), ellagic acid (1.736-1.996 mg/tablet), aloe-emodin (0.337-0.440 mg/tablet), rhein (1.636-2.562 mg/tablet), emodin (0.602-0.846 mg/tablet), chrysophanol (0.388-0.566 mg/tablet) and emodin methyl ether (0.621-0.781 mg/tablet). The quality of the 10 batches of samples was basically the same. Conclusion: This method is simple and accurate and can be used for the quality control of DLP.

Investigation on seeds characteristics and establishment of aseptic cultural system for Rheum palmatum / 中草药

Objective: To study the seeds shapes and germination characteristics, construct aseptic seedlings and callus cultural system, and provide a basis for the rapid propagation and secondary metabolic regulation of Rheum palmatum. Methods: Ten batches of R. palmatum seeds were subjected to characteristic analyses from the aspects of size, purification, weights per thousand seeds, seeds vigor, germination rates, and germination energy. The optimum disinfection system for aseptic seedlings and the optimum hormone ratio for inducing callus were screened by orthogonal test. The content of ten active components in aseptic seedlings and calli were primarily evaluated by HPLC analysis. Results: There was no significant difference in the appearances of the ten batches of R. palmatum seeds from different regions. The content of moisture, seeds vigor, germination rates, and germination energy differed apparently. The germination characteristics in Hezheng and Weiyuan counties were the best. The best disinfection group for aseptic seedlings was the combination of 75% ethanol for 30 s with 10% hydrogen peroxide for 15 min. The optimum hormones for callus induction were 6-BA (1.0 mg/L) + KT (2.0 mg/L) + NAA (1.5 mg/L). Seeds treated with different disinfectants had no significant effect on the content of ten components in germinated aseptic seedlings (P > 0.05). Seven active components were detected in callus, which was significantly lower than that in aseptic seedlings. And the content of chrysophanol-8-O-glucoside was the highest in the callus. Conclusion: The seed characteristics from Hezheng and Weiyuan counties in Gansu Province were excellent by analyses of weights per thousand seeds, seeds vigor, germination rates, and germination energy. The aseptic seedlings and callus cultural system of R. palmatum were successfully established, laying a solid foundation for further study.

Simultaneous determination of eleven constituents in Yigan Yiqi Jieyu Granules by HPLC / 中草药

Objective: To establish HPLC method for the simultaneous determination of saikosaponin a, naringin, paeoniflorin, calycosin-7-glucoside, tanshinone IIA, cinnamaldehyde, schisandrin, syringin, berberine hydrochloride, chrysophanol and hesperidin in Yigan Yiqi Jieyu Granules (YYJG), and conduct a quality assessment using principal component analysis. Methods: The chromatographic separation was achieved on an Caprisil AQ-C18 (150 mm × 4.6 mm, 5.0 μm) column with mobile phase consisted of 0.1% phosphate-acetonitrile for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 45 ℃. The results of the content were then combined with the principal component analysis to achieve the scientific assessment of the different batches of drugs. Results: The content of saikosaponina, naringin, paeoniflorin, calycosin-7-glucoside, tanshinone IIA, cinnamaldehyde, schisandrin, syringin, berberine hydrochloride, chrysophanol and hesperidin in YYJG had good linear relationship in the ranges of 1.6-80.0, 14-700, 10-500, 1.6-80.0, 1.6-80.0, 2.4-120.0, 1.2-60.0, 1.2-60.0, 8.0-400.0, 2.0-100.0, and 2.0-100.0 μg/mL, respectively; The average sample recovery rate range were 98.3%, 99.2%, 98.8%, 99.3%, 101.9%, 97.5%, 99.8%, 101.7%, 101.1%, 102.5%, and 100.9% (RSD < 2.0%); The content of 11 active ingredients in 16 batches of samples respectively were 0.233-0.322, 3.007-3.142, 2.201-2.273, 0.320-0.355, 0.317-0.399, 0.451-0.523, 0.265-0.297, 0.209-0.226, 1.848-1.873, 0.380-0.425, and 0.615-0.647 mg/g, respectively. Conclusion: The established method is simple, accurate and reproducible, and can provide the reference for the quality control of YYJG.

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.

Simultaneous determination of emodin, chrysophanol, parietic acid, and magnolol in Fructus Cannabis Bolus by high performance liquid chromatography / 国际中医中药杂志

Objective@#To simultaneously determinate 4 effective components in Fructus Cannabis Bolus by high performance liquid chromatography (HPLC).@*Methods@#The column was ALLTIMA-C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase was methanol (A)-0.1% phosphoric acid aqueous solution (B) in gradient elution program. The flow rate was 1.0 ml/min, the column temperature was 30℃, and the injection volume was 10 μl. The detection wavelength was set at 254 nm.@*Results@#The linear ranges of 4 effective components of emodin, chrysophanol, parietic acid, and magnolol were 9.05-81.26 μg (r=0.999 8), 13.28-119.3 μg (r=0.997 6), 8.30-75.40 μg (r=0.998 7), 6.30-61.79 μg (r=0.996 5), respectively; the average recoveries and RSDs of emodin, chrysophanol, parietic acid, and magnolol were 99.25%, 99.46%, 99.51%, 99.86%, respectively. The RSDs of emodin, chrysophanol, parietic acid, and magnolol were 2.04%, 3.23%, 1.84%, and 2.42%, respectively.@*Conclusions@#The HPLC can be used as an effective and feasible method for the determination of emodin, chrysophanol, parietic acid, and magnolol in Fructus Cannabis Bolus. The method is simple, quick, and reproducible, and can be used for the quality control of Fructus Cannabis Bolus.

Simultaneous Determination of 7 Components in Niuhuang Qingwei Pills by HPLC-wavelength Switching Method / 中国药房

OBJECTIVE： To develop a method for simultaneous determination of 7 components of Niuhuang qingwei pills as chlorogenic acid， geniposide， forsythoside A， narirutin， baicalin， ammonium glycyrrhetate， chrysophanol. METHODS： HPLC-wavelength switching method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of acetonitrile （A）-0.1％ phosphoric acid （B） gradient elution at the flow rate of 1.0 mL/min. The detection wavelengths were set at 348 nm （chlorogenic acid）， 238 nm （geniposide）， 330 nm （forsythoside A）， 280 nm （narirutin and baicalin）， 237 nm （ammonium glycyrrhetate）， 254 nm （chrysophanol）. The column temperature was 30 ℃， and sample size was 10 μL. RESULTS： The linear ranges of chlorogenic acid， geniposide， forsythoside A， narirutin， baicalin， ammonium glycyrrhetate， chrysophanol were 0.011 67-0.233 4 μg （r＝0.999 4）， 0.042 91-0.858 1 μg （r＝0.999 4）， 0.125 0-2.500 μg （r＝0.999 9）， 0.118 0- 2.360 μg （r＝0.999 9）， 0.119 6-2.392 μg （r＝0.999 7）， 0.030 57-0.611 4 μg （r＝0.999 6）， 0.006 201-0.124 0 μg（r＝0.999 4）， respectively； the limits of quantitation were 1.167， 0.858， 1.250， 1.180， 1.196， 0.611， 0.620 μg/mL， respectively； RSDs of precision tests were 0.98％， 1.04％， 0.59％， 1.50％， 0.83％， 1.24％ and 1.32％ （n＝6）， respectively. RSDs of stability tests were 1.21％， 0.97％， 1.42％， 0.71％， 0.98％， 1.87％ and 1.63％ （n＝6， 12 h）， respectively. Average recoveries were 98.32％， 98.11％， 98.81％， 98.50％， 98.30％， 98.16％ and 97.83％， and the RSDs were 1.37％， 1.41％， 0.64％， 1.01％， 1.18％， 1.16％ and 1.16％ （n＝6）， respectively. CONCLUSIONS： Established method is easy and reproducible. It can be used for the quality control of Niuhuang qingwei pills.

Content Determination of Indicator Components in Jianpi Yishen Pills and the Effects on Calcium ，Phosphorus Metabolism and Inflammatory Factors in CRF Model Rats / 中国药房

OBJECTIVE： To establish the method for the content determination of astragaloside Ⅳ， emodin and chrysophanol in Jianpi yishen pills （JYP） and to investigate the effects of JYP on calcium， phosphorus metabolism and inflammatory factors in chronic renal failure （CRF） model rats. METHODS： HPLC method was adopted. The determination of astragaloside Ⅳ， emodin and chrysophanol was perform on Agilent Zorbax SB-C18， Agilent TC C18 column， respectively； mobile phase consisted of acetonitrile-water （36 ∶ 64， V/V） and methanol-0.1％ phosphoric acid solution （75 ∶ 25， V/V）； the detectors were evaporative light-scattering detector and diode-array detector （detection wavelength of 254 nm）； the column temperatures were set at 30 ℃and 25 ℃ at the flow rate of 1.0 mL/min； the sample sizes were 20 and 10 μL. SD rats were randomly divided into normal group， model group， Niaoduqing group （1.80 g/kg） and JYP low-dose， medium-dose and high-dose groups （1.71， 3.43， 6.85 g/kg）， with 10 rats in each group. Except for normal group， CRF model of other groups were established by 5/6 nephrectomy in other groups. Four months after modeling， normal group and model group were given constant volume of water intragastrically； admi- nistration groups were given relevant medicine intragastrically， once a day， for consecutive 12 weeks. The levels of serum creatinine （Scr）， urea nitrogen （BUN）， parathyroid hormone （PTH） and inflammatory factors （IL-6， TNF-α） were measured by ELISA. Methyl thymol blue colorimetric method and phosphomolybdic acid method were used to detect the contents of blood calcium and phosphorus. Correlation of inflammatory factors with related calcium and phosphorus metabolism indexes （blood calcium， blood phosphorus， PTH） were investigated with Pearson assay. RESULTS： The linear range of astragaloside Ⅳ， emodin and chrysophanol were 54.537-381.759， 2.960-20.720， 6.318-44.223 μg/mL， respectively. The limits of quantitation were 0.010， 0.288， 0.216 μg/mL； the limits of detection were 0.003， 0.096， 0.072 μg/mL. RSDs of precision， reproducibility and stability tests were all lower than 3.0％. The recoveries were 97.18％-102.33％（RSD＜3％，n＝9）. After modeling （before medication）， serum contents of Scr and BUN in model group and administration group were increased significantly， compared with normal group （P＜0.01）. After medication， above indexes of administration group were decreased significantly， compared with model group and the same group before medication （P＜0.01）. Compared with normal group， the content of blood calcium were decreased significantly， while the contents of IL-6 and TNF-α were increased significantly （P＜0.01）. Compared with model group， the content of blood calcium were increased significantly in JYP medium-dose and high-dose groups， while serum content of PTH in Niaoduqing group， serum contents of PTH and IL-6 in JYP medium-dose and high-dose groups as well as serum content of TNF-α in administration group were decreased significantly （P＜0.05 or P＜0.01）. JYP had no significant effect on blood phosphorus in rats， and there was no correlation of inflammatory factors with related calcium and phosphorus metabolism indexes （P＞0.05）. CONCLUSIONS： The established content determination method is simple， specific and sensitive， and can be used for content determination of astragaloside Ⅳ， emodin and chrysophanol in JYP. JYP can improve renal function of CRF model rats， relieve calcium metabolism disorder and inhibit the expression of inflammatory factors.