Analysis on Advantages of New Integration Processing Method in Producing Area of Polygoni Multiflori Radix Praeparata / 中国实验方剂学杂志

ObjectiveTo analyze the effects of new integration processing method in producing area and traditional method on the composition and pharmacological action of Polygoni Multiflori Radix Praeparata（PMRP）， and to illustrate the advantages of toxicity reducing and efficacy enhancing of the decoction pieces prepared by the new method. MethodFresh Polygoni Multiflori Radix（PMR） was taken from Dao-di producing area， and was processed by new integration processing method in producing area（steaming with black bean juice under pressure of 0.1 MPa and temperature at 120 ℃ for 10.5 h） and traditional method（steaming with black bean juice under water for 36 h）， respectively. Samples were collected during the processing process of the two methods， For new method， the samples were collected at 0.5， 3， 5.5， 8， 10.5 h， separately. For traditional method， the samples were collected every 4 h. High performance liquid chromatography（HPLC） was used to establish fingerprint and identify common peaks， the content of polysaccharides was determined by anthrone-sulfuric acid colorimetry at 627 nm， and the contents of anthraquinones and stilbene glycosides in different processed products were determined according to the methods under the item of determination of PMR and PMRP in the 2020 edition of Chinese Pharmacopoeia. In pharmacological experiments， 90 SD rats were randomly divided into 9 groups with 10 in each group（half of male and half of female）， including the blank group， and raw products， 24 h processed products under atmospheric pressure， 30 h processed products under atmospheric pressure， 8 h processed products under high pressure groups with low and high dosages（4.125， 16.5 g·kg-1）. Rats were given the drug by gavage for 29 d with once a day， blood was collected from the abdominal aorta after the last administration， and the serum was isolated， the body mass and liver mass of rats were weighed and the organ index was calculated. The pathological change of liver tissue was observed by hematoxylin-eosin（HE） staining， and biochemical methods were used to detect the contents of aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， alkaline phosphatase（ALP）， γ-glutamyltransferase（GGT）， lactic dehydrogenase（LDH） in serum which used as liver function indicators and the levels of superoxide dismutase（SOD）， malondialdehyde（MDA）， glutathione peroxidase（GSH-Px） in brain tissues which used as oxidation indicators. ResultA total of 14 common peaks were identified in the fingerprint of PMR， PMRP prepared by new method and traditional method， and three of the peaks were designated as stilbene glycoside， emodin and emodin methyl ether， respectively. The characteristic peak areas of each processed products changed significantly from 0 min to 25 min， indicating that different processing methods had an effect on the contents of components with high polarity in PMRP， and the trend of the changes of the two methods was similar， with the higher degree of change in the new method. The determination results showed that compared with the traditional method， the content of polysaccharide（a kind of beneficial component in PMRP obtained by the new method） significantly increased， while the contents of stilbene glycoside and bound anthraquinone（liver-damaging ingredients） significantly decreased. The pharmacological results showed that compared with the blank group， AST and LDH levels of male rats in the low and high dose groups of 24 h processed products under atmospheric pressure and AST level of male rats in the low and high dose groups of 8 h processed products under high pressure were significantly reduced（P<0.05， P<0.01）， while compared with the raw product groups with the same dose， AST and LDH levels of male rats in the low dose group of 30 h processed products under atmospheric pressure were significantly reduced（P<0.05， P<0.01）， the AST levels of male rats in the low and high dose groups of 8 h processed products under high pressure were significantly decreased（P<0.01）， and there was no statistical significance in the differences of biochemical indexes of female rats in each administration group as compared with those of the blank group. ConclusionThe new integration processing method in producing area of PMRP can reach the quality of relevant regulations in 8 h. The processed products obtained by this method have more advantages than the traditional method in terms of toxicity reducing and efficacy enhancing， and energy saving to avoid the loss of ingredients， which can provide ideas for the production of high-quality decoction pieces of PMRP， and the integration processing method in producing area of other roots and rhizomes of traditional Chinese medicines.

Simultaneous determination of 10 rhubarb anthraquinones and rhaponticin in Compound gentian and sodium bicarbonate tablets by UPLC / 中国药房

OBJECTIVE To establish a method for the simultaneous determination of 10 rhubarb anthraquinones in Compound gentian sodium bicarbonate tablets and the content of rhaponticin，which are the characteristic components of artifacts，and to use the method to evaluate the quality of compound preparation containing Rheum officinale. METHODS The ultra-performance liquid chromatography （UPLC） method was adopted to determine the contents of 10 rhubarb anthraquinones （aloe-emodin-8-O-glucoside， rheinic acid-8-O-β-D-glucoside，emodin-8-O-glucoside，chrysophanol-8-O-β-D-glucoside，emodin monomethyl ether-8-O-β-D-glucoside， aloe-emodin，rheinic acid，emodin，chrysophanol，emodin monomethyl ether） and rhaponticin in 40 batches of Compound gentian sodium bicarbonate tablets from 8 manufacturers. The determination was performed on the Agilent Eclipse Plus C18 column with a mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution） at a flow rate of 0.3 mL/min； the column temperature was set at 30 ℃ ，and the injection volume was 5 μL. Combining principal component analysis and cluster analysis to synthesize the results of content determination，the quality of samples from different manufacturers was evaluated. RESULTS All of above 11 components showed favorable linear relationships with peak areas in their respective mass concentration ranges （r≥0.999 3），with RSDs of precision，repeatability and stability 296261547@qq.com less than 3% （n＝6）； average sample recoveries ranged 96.82%-98.92% （RSD≤1.74%，n＝6）； their contents were 0971-8247794。E-mail：304436784@qq.com 0.011 7-0.252 0，0-0.323 3，0.131 3-1.236 6，0.081 1-1.056 2，0.015 2-0.189 8，0.001 8-0.152 3，0-0.255 2，0.001 9-0.223 4，0.054 3-0.303 0，0.022 7-0.172 2，0-2.835 9 mg/g，respectively. The cumulative variance contribution of the first three principal components was 95.533%； the 40 batches of samples can be clustered into 4 categories：samples from enterprises a and d were in a category of their own，samples from enterprises f，b，g and e were in a category，and samples from enterprises c and h were in a category. There were large differences in the content of rhubarb anthraquinone in the samples from 8 manufacturers，and rhaponticin was only detected in the sample from one enterprise. CONCLUSIONS Established UPLC method is stable and reliable； it can be used for the content determination of 10 rhubarb anthraquinones and rhaponticin in Compound gentian sodium bicarbonate tablets.

Phytotoxicity of plant extracts of Vismia japurensis cultivated in vivo and in vitro

Abstract Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.

Resumo Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.

Phytotoxicity of plant extracts of Vismia japurensis cultivated in vivo and in vitro / Fitotoxicidade de extratos de plantas de Vismia japurensis cultivadas in vivo e in vitro

Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.

Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.

Phytotoxicity of plant extracts of Vismia japurensis cultivated in vivo and in vitro / Fitotoxicidade de extratos de plantas de Vismia japurensis cultivadas in vivo e in vitro

Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.

Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.

Study on the quality standard of Kuipingning gastric floating tablets / 中国药房

OBJECTIVE To establish the quality standard of Kuipingning gastric floating tablets. METHODS Kuipingning gastric floating tablets were prepared and investigated in terms of property ，weight difference and friability. Crydalis yanhusuo was identified qualitatively by thin layer chromatography （TLC）method. High performance liquid chromatography method was used to determine the content of total anthraquinones in Rheum palmatum ，and set the content limit of total anthraquinones. The floating performance and release degree of the preparation were investigated ，and the release kinetic process was fitted. RESULTS Kuipingning gastric floating tablets prepared in this study were gray white to gray tablets with slight smell and bitter taste ；the weight difference and friability were all in line with relevant regulations ；the established TLC method possessed strong specificity and could accurately identify C. yanhusuo . The average content of total anthraquinones in R. palmatum was 17.95 mg/tablet，and its content limit would not be less than 14.36 mg/tablet. The initial floating time of the preparation was no more than 10 s，and the holding time was more than 8 h. The release kinetics process accorded with the Retger-Peppas release model. CONCLUSIONS The method established in this study shows good reliability ，stability and feasibility ，and can effectively control the quality of Kuipingning gastric floating tablets.

Investigation of Metabolic Stability of Phase Ⅰ and Ⅱ Sulfation of Lucidin in Liver Microsomes and Liver S9 from Different Species / 中国实验方剂学杂志

ObjectiveTo evaluate the metabolic stability of lucidin by incubating liver microsomes and liver S9 from 4 species， and to compare the species differences in metabolism of lucidin in vitro. MethodA qualitative and quantitative method of lucidin based on ultra-high performance liquid chromatography-high resolution mass spectrometry （UHPLC-HRMS） was established and verified. Lucidin was incubated with rat， mouse， beagle dog， human liver microsomes and liver S9 to investigate the metabolic stability parameters， metabolites， metabolic pathways. ResultHepatic clearance （CLh） of lucidin was in order of mouse>rat>beagle dog>human in both phase Ⅰ and phase Ⅱ incubation system. Its metabolic stability was good in rat， beagle dog and human， while it showed metabolic instability and moderate metabolic stability in mouse microsomes and liver S9， respectively. A total of 5 metabolites were rapidly identified， including 3 oxidation metabolites of phase Ⅰ and 2 sulfation metabolites of phase Ⅱ. The production rate of metabolites was consistent with the results of metabolic stability. ConclusionThe established UHPLC-HRMS is simple and specific， which can be used for the study on the metabolic stability and metabolites of lucidin. Its metabolic stability and metabolite production rate in vitro are significantly different among species， the metabolic characteristics of rat and beagle dog are similar to human， which provides an important reference for subsequent research in vivo， safety evaluation and animal model selection of lucidin.

Organ specific differences in alteration of aquaporin expression in rats treated with sennoside A, senna anthraquinones and rhubarb anthraquinones / 中国药理学与毒理学杂志

OBJECTIVE Senna and rhubarb are classified as stimulative laxatives, and known to have similar effec?tive constituents, the anthraquinones. Being protected by theβ-glucoside bond, the anthraquinones can reach the intes?tines where they are degraded into complex metabolites by enzymes secreted from the intestinal microbiome. It is these complex metabolites that produce the laxative effects. Then the similarities and differences of action between the anthra?quinones require further elucidation. METHODS Here, we studied metabolites of senna anthraquinones (SAQ), rhubarb anthraquinones (RAQ) and their chemical marker, sennoside A (SA), in a rat diarrhea model. In the in vitro biotransfor?mation experiments, SAQ, RAQ and SA were incubated with rat fecal flora solution and the metabolites produced were analyzed using HPLC. In the in vivo studies, the same compounds were investigated for purgation induction, with mea?surement of histopathology and multiple aquaporins (Aqps) gene expression in six organs. RESULTS SAQ and RAQ had similar principal constituents but could be degraded into different metabolites. A similar profile of Aqps down-regula?tion for all compounds was seen in the colon, suggesting a similar mechanism of action for purgation. However, in the kidneys and livers of the diarrhea-rats, down-regulation of Aqps was found in the RAQ-rats whereas up-regulation of Aqps was seen in the SAQ-rats. Furthermore, the RAQ-rats showed lower aquaporin 2 (Aqp2) protein expression in the kidneys, whilst the SA-rats and SAQ-rats had higher Aqp2 protein expression in the kidneys. This may have implications for side effects of SAQ or RAQ in patients with chronic kidney or liver diseases. CONCLUSION SAQ and RAQ showed similar laxative actions with a similar mechanism, they could display different actions in rat kidneys and livers. We suggest that the clinical usage of senna or rhubarb products should be clarified for patients having chronic kidney or liver diseases.

Analysis on Chemical Constituents in Xiao Chengqitang by UPLC-Q-Orbitrap-MS / 中国实验方剂学杂志

Objective:To rapidly identify the chemical constituents in Xiao Chengqitang by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS）. Method:The method was established by the Waters CORTECS T3 column （2.1 mm×150 mm， 1.6 μm）， mobile phase was methanol （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-5 min， 3%-21%A； 5-20 min， 21%-36%A； 20-32 min， 36%-50%A； 32-42 min， 50%-62%A； 42-50 min， 62%-85%A； 50-60 min， 85%-95%A）， the flow rate was 0.2 mL·min^-1， and the column temperature was 30 ℃. UPLC-Q-Orbitrap-MS was operated in positive and negative ion modes， the scanning range was 100-1 200 with mode of Full MS/dd-MS²， and the collision energies were 20， 40 eV. The compounds were identified by comparing with reference substances and combining with literature reports and MS database information. Result:A total of 123 components were identified in Xiao Chengqitang， including 33 flavonoids， 25 anthraquinones and anthrones， 23 phenylpropanoids， 15 tannins， 10 nitrogen-containing components and 17 other components. Among them， 32 components were determined by reference substances. Conclusion:The material basis of Xiao Chengqitang is flavonoids， anthraquinones and anthrones， phenylpropanoids， which is derived from Aurantii Fructus Immaturus，<italic> </italic>Rhei Radix et Rhizoma and Magnoliae Officinalis Cortex， respectively.

Research progress of effect of Rhei Radix et Rhizoma and its anthraquinone in treatment of autoimmune diseases / 中国中药杂志

Rhei Radix et Rhizoma was first recorded in Shennong Ben Cao Jing, with a wide range of pharmacological activities. Autoimmune disease is a kind of disease that damages the tissue structure and function of immune cells and their components due to the impairment of immune tolerance function, including atherosclerosis, multiple sclerosis, gout, rheumatoid arthritis, autoimmune thyroiditis, ulcerative colitis, type 1 diabetes and IgA nephropathy. In recent years, clinical and experimental studies show that Rhei Radix et Rhizoma has potential therapeutic effects on autoimmune diseases. Under the guidance of the theory of traditional Chinese medicine, this paper reviews therapeutic and intervening effects of Rhei Radix et Rhizoma and its main active ingredient anthraquinone on autoimmune diseases. It also puts forward new study directions in view of the existing problems in studies of rhubarb and its anthraquinone, with the aim to provide reference for clinical treatment and scientific studies of effect of Rhei Radix et Rhizomaon autoimmune diseases.

Exploring Research on Processing Method of Polygoni Multiflori Radix Praeparata / 中国实验方剂学杂志

Objective::To investigate the effects of black bean juice with different stewing times on the appearance character and the content changes of effective components of Polygoni Multiflori Radix Praeparata. Method::HPLC was employed with Agilent ZORBAX Extend-C18 column (4.6 mm×250 mm, 5 μm), a gradient mobile phase of methanol (A)-water (B) was eluted (0-30 min, 5%-100%A; 30-40 min, 100%A), the flow rate was 1.0 mL·min-1, the injection volume was 10 μL and the column temperature was 35 ℃, detection wavelength was set at 280 nm. The contents of stilbene glycoside, emodin, emodin methyl ether, emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside in samples prepared at different processing times were simultaneously determined by HPLC. Result::The content of stilbene glycoside decreased gradually with the increase of stewing time, compared with 8 h, its content decreased by 76% at 64 h. The contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside increased first, and then decreased, reaching the highest value at 24 h, and then decreased to the level similar to the content of 8 h after 40 h, and then fluctuated slightly. The contents of emodin and emodin methyl ether increased first, and then decreased, reached the maximum when stewed for 32 h, then decreased slowly and tended to be stable. Conclusion::The stewing time has significant influence on the content of various components in Polygoni Multiflori Radix Praeparata, and the changing trend is different, the processing time of Polygoni Multiflori Radix Praeparata shall be standardized. At the same time, it is not sufficient to take stilbene glycoside and anthraquinones as the indicator ingredients for this decoction pieces, the quality control indicators such as polysaccharides shall be considered.

Analysis of Chemical Composition in Sancao Baogan Decoction Based on UPLC-ESI-HRMSn / 中国实验方剂学杂志

Objective::To analysis the chemical constituents in Sancao Baogan decoction by ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-ESI-HRMSn). Method::The separation was performed on an ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) by a gradient elution of methanol (A)-0.1% formic acid solution (B) (0-2 min, 5%A; 2-20 min, 5%-12%A; 20-35 min, 12%-40%A; 35-38 min, 40%A; 38-48 min, 40%-80%A; 48-50 min, 80%A). The flow rate was 0.3 mL·min-1 and the column temperature was set at 30 ℃, the injection volume was 10 μL. Electrospray ionization was applied and the data were collected via positive and negative ion modes. By using Xcalibur 3.0 software, the chemical constituents were analyzed based on the relative retention time, excimer ion peak and fragment ion peak of the compounds, as well as comparison with human metabolome database (HMDB), reference substances and literature data. Result::A total of 40 chemical components were identified from Sancao Baogan decoction, including 16 phenolic acids, 19 flavonoids, 2 anthraquinones, 1 triterpenoid, 1 sterol, and 1 monoterpenoid. Six compounds (dansensu, α-pinene, epigallocatechin, 2, 5-dihydroxybenzoic acid, naringenin and emodin) were reported for the first time from Sancao Baogan decoction. Conclusion::The established UPLC-ESI-HRMSn can quickly, accurately and comprehensively analyze the chemical constituents of Sancao Baogan decoction, which lays a foundation for the basic research of pharmacodynamic substances and quality control of this formula.

Research progress on synthesis of anthraquinones based on shikimic acid/o-succinylbenzoic acid pathway and polyketone pathway / 中草药

Anthraquinone and its derivatives are very important secondary metabolites in plants. They have many functions such as photoprotection and improvement of plant disease resistance. They also have very important applications in the fields of medicine and chemical engineering. Efficiently and quickly obtaining anthraquinones and improving the synthesis efficiency of anthraquinones in plants have become one of the research focuses of modern synthetic biology. However, the synthetic pathway of anthraquinones is more complicated. At present, it is generally believed that anthraquinones are formed in plants by the shikimic acid/o- succinylbenzoic acid pathway and polyketone pathway. This article focuses on the recent research advances in the skeleton synthesis of anthraquinone via shikimic acid/o-succinylbenzoic acid pathway and polyketone pathway in plants, and provides a certain theoretical basis for studying the synthesis and regulation of anthraquinone metabolites in plants.

Simultaneous determination of four anthraquinones in Polygonum multiflorum by QAMS / 中国中药杂志

A simple, specific and selective quantitative analysis of multi-components by single marker(QAMS) method for simultaneous determination of anthraquinones and anthraquinone glycosides in Polygonum multiflorum was developed. Four main anthraquinones and its glycosides, emodin, emodin-8-O-β-D-glucoside, physcion and physcion-8-O-β-D-glucoside were selected as analytes to evaluate the quality of P. multiflorum. Emodin was used as the internal standard, and the relative correction factors(RCFs) between emodin and the other three anthraquinones were calculated. Comparison of the contents of the four components in 30 batches of P. multiflorum from different regions and 12 batches decoction pieces from different manufacturers by QAMS and external standard method(ESM) showed that there was no significant difference between QAMS and ESM for quantification of the four main components by using relative error results, and the QAMS method was accurate and reliable, and had a good repeatability. In addition, compared with the results calculated by the difference method between total anthraquinone and free anthraquinone in the content determination of P. multiflorum in Chinese Pharmacopoeia, the results of direct determination combined anthraquinone by QAMS were very close to that by measured the external standard method. Therefore, simultaneous quantification of four main anthraquinones by using QAMS is suitable to evaluate the quality of P. multiflorum. Then the optimized assay method of the combined anthraquinone contents showed simple and feasible, which could be replaced and improved the quantification method of the combined anthraquinone in the current Chinese Pharmacopeia.

Phytochemistry and fungitoxic potential of extract and fractions of pouteria ramiflora on fusarium solani f. sp. phaseoli / Fitoquímica e potencial fungitóxico de extrato e frações de Pouteria ramiflora SOBRE Fusarium solani f. sp. phaseoli

Among the fungus that attack the common bean crop there is Fusarium solani f.sp. phaseoli (Fsp), causing major losses. Taking into account its importance, the objective of this work was to determine the secondary metabolites class and the potential fungicide of the Pouteria ramiflora leaves on Fsp. The ethanol extract was dissolved in methanol/water and partitioned successively with hexane, dichloromethane, chloroform, ethyl acetate and butanol, subjected to the chemical profile. The solutions were prepared in concentrations of 800, 1200, 1600, 2000 and 2400 µ g/mL and poured into Petri dishes; then, 0.5 cm disc of diameter with spores and mycelia of Fsp was deposited. These dishes were incubated in temperature of 25 ± 2 °C and the evaluations, performed by measuring the colonies diameter (five replications) until reaching the dishes border (three days) with a completely randomized experimental design. Through the Mycelial Index Growth Speed (MIGS) data, analysis of variance was performed and when significant, applied the regression analysis. The results indicated that all fractions and the extract have the phenolic compounds and/or derivatives as one of the major constituents except the hydroethanolic fraction. The extract and its fractions decreased the Fsp MIGS, in the same proportion in which concentrations were increased; the greatest reduction occurred in the butanol fraction at the concentration of 2400 µg/mL, with growth close to zero, indicating its potential use for the Fsp control, probably due to the presence of anthraquinones.

Entre os fungos que acometem a cultura do feijão está Fusarium solani f.sp. phaseoli, causando perdas na produtividade. Levando-se em consideração sua importância, o objetivo deste trabalho foi determinar a classe de metabólitos secundários e o potencial fungicida das folhas de curriola (Pouteria ramiflora) sobre F. solani, em condições de laboratório. O extrato etanólico foi dissolvido em metanol/água e particionado sucessivamente com hexâno, diclorometano, clorofórmio, acetato de etila e butanol, submetidos ao perfil químico. As soluções foram preparadas nas concentrações de 800, 1200, 1600, 2000 e 2400 µ g/mL evertidas em placas de petri (10 mL); em seguida, foi depositado um disco de 0,5 cm de diâmetro com esporos e micélio de F. solani. As placas foram incubadas a temperatura de 25 ± 2 °C e as avaliações, realizadas por meio da medição do diâmetro das colônias (cinco repetições) até atingir a borda da placa (três dias), com delineamento experimental inteiramente casualizado. Através dos dados do Índice de Velocidade de Crescimento Micelial (IVCM), foi realizada a análise de variância e quando significativa, aplicada a análise de regressão. Os resultados indicaram que todos as frações e o extrato possuem os compostos fenólicos e/ou derivados como um dos constituintes majoritários, exceto a fração hidrometanólica. O extrato e suas frações diminuíram o IVCM de F. solani, à medida que se aumentava as concentrações; a maior redução ocorreu na fração butanolica para a concentração de 2400 µ g/mL, com crescimento próximo a zero, indicando seu potencial de uso para controle de F. solani, provavelmente devido a presença de antraquinonas em sua composição.

Anti-cancer Effect of Anthraquinones in Polygoni Multiflori Radix / 中国实验方剂学杂志

Objective: Polygoni Multiflori Radix is the drought root of a perennial vine, Polygonum multiflorum, which is belong to Polygonaceae. According to Chinese Pharmacopeia (edition 2015), its preparations are divided into two types based on the processing:crude Polygoni Multiflori Radix, and processed Polygoni Multiflori Radix Praparata. This study aimed to explore the substantial bases, effect and the mechanisms of anthraquinones in Polygoni Multiflori Radix. Method: Based on the literatures retrieved in PubMed, CNKI and Chongqing VIP in past 15 years, the chemical composition, pharmacological effects and mechanisms of anthraquinones in Polygoni Multiflori were reviewed and summarized. Result: There were 20 anthraquinones in Polygoni Multiflori. Compared with other Polygonaceae plants, there were higher contents of emodin and physcion, both were qualification markers in Polygoni Multiflori. In terms of the anti-cancer effect, emodin, emodin-8-O-β-D-glucoside, aloe-emodin, chrysophanol, physcion, physcion-8-O-β-D-glucoside and rhein induced cancer cell apoptosis. Emodin, aloe-emodin, chrysophanol, physcion and rhein inhibited cancer cell cycles. Emodin, aloe-emodin, physcion and emodin-8-O-β-D-glucoside blocked migration, invasion and metastasis. And emodin, chrysophanol and rhein blocked energy metabolism, and emodin reversed multidrug resistance in cancer. Conclusion: Polygoni Multiflori may play a potential anti-cancer value, and its underlying mechanisms and the interaction between components are worth of further studies.

Research on contents of anthraquinones,dianthrones and tannins in Rheum tanguticum on PCA and CA / 中国中药杂志

Anthraquinones,dianthrones and tannins are the main active ingredients of Rheum tanguticum. In this study the three components were determined by HPLC,and the results were analyzed by multiple comparisons,principal components analysis(PCA)and correspondence analysis(CA). The results showed that the contents of components in different growing areas and types(wild and cultivated) reached a significant level(P<0. 05). Baiyu county,Xiaojin county and Ruoergai county had obvious advantages in the accumulation of catechin hydrate,rhien and sensenoside A respectively. The principal component was different in two growing type and the wild environment was conducive to combined anthraquinones accumulation. For active components,normalized planting was better than retail cultivating. Therefore,the effect on the accumulation of chemical components in Rh. tangusticum,should be taken into full account in the selection of the cultural base of Rh. tanguticum. The standardized cultivating is superior to retail cultivating in terms of the accumulation of active ingredients,and standardized planting is inferior to the wild.

Evaluation of chemical quality profile of Polygoni Multiflori Radix at different processing degrees based on its classic processing method "nine-steaming and nine-sun-curing" / 中国中药杂志

Based on the ancient method of nine-steaming and nine-sun-curing,the chemical composition changes and quality profiles in different processes of Polygoni Multiflori Radix were studied. Their contents of stilbene glycoside,anthraquinones and polysaccharides were determined by nine-steaming and nine-sun-curing with black bean juice and pharmacopoeia method. HPLC chemical fingerprints were established,and orthogonal partial least squares-discriminant analysis( OPLS-DA) was performed on different processed products using SIMCA 14. 1 software to evaluate the quality difference between samples. The results of content determination show that,with the increase of the number of processing and steaming times,the stilbene glycoside and the combined anthraquinone showed a decreasing trend,and the free anthraquinone,total anthraquinone and polysaccharide showed an upward trend in the different preparations of Polygoni Multiflori Radix and Pharmacopoeia. Six-steamed and six-sun-cured products can be used as the finishing point for the classic steaming. Fingerprint results showed that there were significant differences in chemical composition in Polygoni Multiflori Radix at different processing processes. It can be identified stilbene glycoside( peak 13),emodin( peak 21),and physcion( peak 24). By comparing the relative peak areas of the 26 chromatographic peaks in the sample after normalization( the reference is peak 7),it was found that the relative peak areas of 12 peaks in the processed products were higher than the raw products,13 peaks were reduced; according to statistical analysis of OPLS-DA,Polygoni Multiflori Radix at different processing degrees was further divided into three categories,sample S1 was class I,S2-S5 were class Ⅱ,and S6-S11 were class Ⅲ． And 8 peaks with the VIP value higher than 1. 0 were peak 13,21,4,3,11,14,5,and 24 in order. The eight chemical components were the main components to distinguish the difference between Polygoni Multiflori Radix in the process of nine-steaming and nine-sun-curing,suggesting that it was rational to use stilbene glycoside,emodin and emodin methyl ether as quality control indicators of Polygoni Multiflori Radix. The method established in this experiment conformed to the methodological verification requirements,established a method of multi-component content determination combined with fingerprint,and clarified that six-steaming and six-sun-curing was used as an improved classical processing technology,and more clearly defined the whole dynamic change of chemical composition in Polygoni Multiflori Radix by nine-steaming and ninesun-curing process. It provides a basis for the chemical quality evaluation model about different processed products of Polygoni Multiflori Radix.

Design and evaluation of rhubarb total free anthraquinones oral colon-specific drug delivery granules to improve the purgative effect

Rhubarb is commonly used as a cathartic in Asian countries. However, researchers have devotedextensive concerns to the quality control and safety of rhubarb and traditional Chinese preparations composed of rhubarb due to the instable purgative effect and potential nephrotoxicity of anthraquinones. In this study, we aimed to prepare rhubarb total free anthraquinones (RTFA) oral colon-specific drug delivery granules (RTFA-OCDD-GN) to delivery anthraquinones to colon to produce purgative effect. RTFA-OCDD-GN were prepared using chitosan and Eudragit S100 through a double-layer coating process and the formulation was optimized. Continuous release studies were performed in a simulated gastric fluid (pH 1.2), followed by a small-intestinal fluid (pH 6.8) and a colonic fluid (pH 7.4, containing rat cecal contents). The purgative effect test was performed in rats. The dissolution profile of RTFA-OCDD-GN showed that the accumulative dissolution rate of RTFA was about 83.0% in the simulated colonic fluid containing rat cecal contents and only about 9.0% in the simulated gastrointestinal fluids. And the RTFA-OCDD-GN could produce the comparative purgative activity as rhubarb, suggesting it could deliver the free AQs to the colon. The RTFA-OCDD-GN was a useful media to enhance the purgative activity of free anthraquinones after administered orally.

Quality evaluation of rhubarb pieces by hplc fingerprint, q-tof/ms qualitative and multi-component quantitative analysis / 中草药

Objective To establish a more comprehensive quality evaluation method for rhubarb pieces. Methods Fingerprints of 35 batches of rhubarb pieces were determined by HPLC, the control fingerprint was established, the common peaks were calibrated and the similarity was evaluated. Components of common peaks were identified by Q-TOF/MS and the content of 13 anthraquinones confirmed by reference substance was determined. Results There were 45 common peaks in the fingerprint of rhubarb pieces, including 17 anthraquinones (among them, eight bound anthraquinones of aloe-emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe-emodin-3-hydroxymethyl-O-glucoside, emodin-8-O- glucoside, physcion-8-O-glucoside and five free anthraquinones of aloe-emodin, rhein, emodin, chrysophanol, physcion were confirmed by reference substance), two anthrones (sennoside B and sennoside A, confirmed by reference substances), 17 tannins (among them, gallic acid, catechin, epigallocatechin gallate, epicatechin and epicatechin gallate were confirmed by reference substance), two stilbenes [resveratrol 4′-O-glucoside and resveratrol 4′-O-β-D-(6″-O-galloyl)-glucoside, confirmed by reference substances], four butyrophenone [among them, 4-(4′-hydroxyphenyl)-2-butanone 4′-O-β-D-(2″-O-galloyl-6″-O-p-hydroxy-cinnamyl) glucoside, confirmed by reference substances], two chromones and one naphthalenes. The content of 13 anthraquinones in 35 batches of rhubarb pieces was quite different. Conclusion The established method can be used to evaluate the quality of rhubarb pieces.