Discovery of human pancreatic lipase inhibitors from root of Rhodiola crenulata via integrating bioactivity-guided fractionation,chemical profiling and biochemical assay / 药物分析学报

Although herbal medicines(HMs)are widely used in the prevention and treatment of obesity and obesity-associated disorders,the key constituents exhibiting anti-obesity activity and their molecular mechanisms are poorly understood.Recently,we assessed the inhibitory potentials of several HMs against human pancreatic lipase(hPL,a key therapeutic target for human obesity),among which the root-extract of Rhodiola crenulata(ERC)showed the most potent anti-hPL activity.In this study,we adopted an integrated strategy,involving bioactivity-guided fractionation techniques,chemical profiling,and biochemical assays,to identify the key anti-hPL constituents in ERC.Nine ERC fractions(retention time=12.5-35 min),obtained using reverse-phase liquid chromatography,showed strong anti-hPL activity,while the major constituents in these bioactive fractions were subsequently identified using liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-Q-TOF-MS/MS).Among the identified ERC constituents,1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose(PGG)and catechin gallate(CG)showed the most potent anti-hPL activity,with pIC50 values of 7.59±0.03 and 7.68±0.23,respectively.Further investigations revealed that PGG and CG potently inhibited hPL in a non-competitive manner,with inhibition constant(Ki)values of 0.012 and 0.082 μM,respectively.Collectively,our integrative analyses enabled us to efficiently identify and characterize the key anti-obesity constituents in ERC,as well as to elucidate their anti-hPL mechanisms.These findings provide convincing evidence in support of the anti-obesity and lipid-lowering properties of ERC.

Determination of catechins in cosmetics by high⁃performance liquid chromatography / 上海预防医学

ObjectiveTo establish a high⁃performance liquid chromatography （HPLC） quantitative method for the determination of epicatechin （EC）， （-）⁃epicatechin gallate （ECG）， （-）⁃epigallocatechin （EGC）， （-）⁃epigallocatechin gallate （EGCG）， （-）⁃gallocatechin （GC）， （-）⁃gallocatechin gallate （GCG）， （-）⁃catechin gallate （CG）， cianidanol （CD） and gallic acid （GA） in cosmetics. MethodsSamples were prepared by ultrasonic extraction and followed by high-speed centrifugation of the extraction solution. The supernatant was filtered by 0.45 μm Millipore filter. The continued filtrate was taken for analysis. A reversed phase column， Kromasil 100-5 C18 （5 μm， 4.6 mm×250 mm） was used with 0.05% trifluoroacetic acid buffer and methanol as mobile phase under the condition of gradient elution. Diode array detection （DAD） method was used for the determination. Qualitative and quantitative determination was conducted in 10 batches of commercially available cosmetics. ResultsThe relative standard deviations （RSD） were in the range of 0.11%-6.30% （n=3）； the recoveries were in the range of 84.4%-114.7%. The method showed a good linearity within the concentration range of 0.49-105.39 mg·L-1 （r>0.995）. The detection limit was 5 μg·g-1. In 10 batches of commercially available cosmetics， three batches showed positive result， which was consistent with the UV spectrum of the standard. ConclusionThis method is efficient， sensitive and accurate. It is applicable to the determination of EC， ECG， EGC， EGCG， GC， GCG， CG， CD and GA in cosmetics.

Ellagic acid production using polyphenols from orange peel waste by submerged fermentation

Background: Biotechnological processes are part of modern industry as well as stricter environmental requirements. The need to reduce production costs and pollution demands for alternatives that involve the integral use of agro-industrial waste to produce bioactive compounds. The citrus industry generates large amounts of wastes due to the destruction of the fruits by microorganisms and insects together with the large amounts of orange waste generated during the production of juice and for sale fresh. The aim of this study was used orange wastes rich in polyphenolic compounds can be used as source carbon of Aspergillus fumigatus MUM 1603 to generate high added value compounds, for example, ellagic acid and other molecules of polyphenolic origin through submerged fermentation system. Results: The orange peel waste had a high concentration of polyphenols, 28% being condensed, 27% ellagitannins, 25% flavonoids and 20% gallotannins. The major polyphenolic compounds were catechin, EA and quercetin. The conditions, using an experimental design of central compounds, that allow the production of the maximum concentration of EA (18.68 mg/g) were found to be: temperature 30°C, inoculum 2 × 107 (spores/g) and orange peel polyphenols 6.2 (g/L). Conclusion: The submerged fermentation process is an effective methodology for the biotransformation of molecules present in orange waste to obtain high value-added as ellagic acid that can be used as powerful antioxidants, antibacterial and other applications.

Antibacterial substances from leaves of Protium spruceanum (Burseraceae): in vitro and in silico evaluation

Due to the increase of bacterial resistance, the search for new antibiotics is necessary and the medicinal plants represent its most important source. The aim of this study was to evaluate the antibacterial property of extract and fractions from Protium spruceanum leaves, against pathogenic bacteria. By means of diffusion and microdilution assays, the crude extract was active against the nine bacteria tested being the hydromethanolic fraction the most active. During phytochemical procedures, procyanidin (1) and catechin (2) were identified as the main antibacterial constituents of this fraction. In silico results obtained using PASSonline tool indicated 1 and 2 as having good potential to interact with different targets of currently used antibiotics. These results no indicated potential to none DNA effect and indicated the cell wall as mainly target. Electrophoresis result supported that had no DNA damage. Cell wall damage was confirmed by propidium iodide test that showed increased membrane permeability and by cell surface deformations observed in scanning electronic microscopy. The in vitro assays together with the in silico prediction results establish the potential of P. spruceanum as source of antibacterial compounds that acts on important bacterial targets. These results contribute to the development of natural substances against pathogenic bacteria and to discovery of new antibiotics.

Identification and comprehensive quality evaluation of Cynomorium songaricum in different producing areas based on OPLS combined with entropy weight TOPSIS method / 中草药

Objective: To identify and comprehensively evaluate Cynomorium songaricum from different producing areas in order to provide reference for the quality evaluation of C. songaricum and the determination of the suitability of the origin. Methods: A total of 40 samples from five provinces (regions) were collected to measure the content of gallic acid, protocatechuic acid, catechins, total polysaccharides, total flavonoids, Na, K, Ca, Mg, Fe, Zn, Mn, Co, Sr, Ni, Ag, Ba, Ti, Cu, Pb, Cr, Cd, As, and Hg. The data reflecting the quality of C. songaricum were analyzed by orthogonal partial least squares discriminant analysis (OPLS-DA) and entropy weight TOPSIS analysis. Results: The contents of Mn, Zn, Co, catechin, Pb, Cr, Ca, Ti, total flavonoids, protocatechuic acid, Mg and Cu in C. songaricum are important for distinguishing different producing areas. The quality of C. songaricum in Inner Mongolia was the best in all provinces (regions), followed by Gansu, Ningxia, Xinjiang, and Qinghai Provinces. Conclusion: The results of OPLS-DA combined with entropy weight TOPSIS analysis are reasonable, objective and effective, and can be applied to the comprehensive evaluation of multiple indicators of C. songaricum.

Dissolution rule of Siwu Decoction by different decocting methods based on chemistry holography / 中草药

Objective: To study the content variation and chemical composition of Siwu Decoction between mixed decoction and single decoction comprehensively, and then explore variation rule of Siwu Decoction by different decocting methods based on material basis. Methods: Components of Siwu Decoction were identified by LC-MS/MS and an UPLC wavelength switching method for simultaneously determining the contents of multiple compounds in Siwu Decoction was established based on the idea of TCM chemistry holography. The mixed and single decoction samples were prepared and tested. Experimental data was compared to analyze material basis differences and variation rule of Siwu Decoction by different decocting methods. Results: A total of 72 compounds were identified and assigned, 18 compounds were quantitative detected and all of 18 analytes showed good linearity (R2 ≥ 0.999) within the test range. The relative standard deviations of the precision, repeatability and stability were not exceeding 2.0%, and the recoveries were in the range of 97%-105%. Analysis of Siwu Decoction samples showed dissolution of ligustilide, 3-n- butylphthalide, catechin, gallic acid and paeoniflorin was affected by the change of solvent volume and dissolution of aucubin, catechin, oxypaeoniflorin, paeoniflorin and acteoside were higher in mixed decoction than single decoction obviously. Compared to single decoction, the kinds of compounds in mixed decoction did not change significantly but the content showed notable variety. Conclusion: Through the study of chemistry holography, the composition and content of compounds in TCM mixed decoction and herbs single decoction can be compared and analyzed comprehensively to provide a new perspective for the study on the rule of TCM decoction and dissolution. TCM chemistry holography study may become a useful exploration of the TCM quality study.

Mechanism of Wuling Powder in treatment of chronic heart failure based on network pharmacology / 中草药

Objective: To explore the mechanism of Wuling Powder in the treatment of chronic heart failure (CHF) based on network pharmacology. Methods: Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) and literature mining were used to search the chemical components and targets of Wuling Powder, and a single drug-active ingredients-target network was established. The related targets of chronic heart failure were collected through Genecards and OMIM databases, the network model of active components-CHF-targets was constructed and analyzed by Cytoscape 3.7.1 software. The protein-protein interaction (PPI) network was constructed by STRING database platform, and the gene oesthetics (GO) function annotation and Kyoto Encyclopedin of Genes and Genomes (KEGG) signal pathway enrichment analysis were performed by DAVID online tools. The molecular docking was performed using Surflex software. Results: Fifty components, 29 potential targets, 8 243 targets related to chronic heart failure, and 27 targets of Wuling Powder-CHF were obtained. The network analysis results showed that the key targets of Wuling Powder in the treatment of chronic heart failure included CASP3, RELA, AR, ESR1, CHRM1 and CASP8, etc. Biological processes mainly involved signal transduction, nervous system development, transcription initiation from RNA polymerase II promoter in response to estradiol, synaptic transmission, cholinergic synaptic transmission, etc. KEGG enrichment involved neuroactive ligand-receptor interaction, PI3K-Akt signaling pathway, cholinergic synapse, etc. The molecular docking results showed that (+)-catechin, taxifolin and other key compounds in Wuling Powder had better binding ability with key targets such as CASP8, CHRM1, and NR3C1. Conclusion: The material basis and mechanism of Wuling Powder in the treatment of chronic heart failure were revealed based on network pharmacology, which provided a certain theoretical basis for the clinical application of Wuling Powder.

Quality study on Liuwei Dihuang series preparations via network pharmacology and active ingredient determination / 中草药

Objective: According to the thinking of research on quality control of traditional Chinese medicine based on “components reflect activity, activity points to efficacy”, the active ingredients obtained through network pharmacology screening were used as evaluation indicators to conduct a re-evaluation study on Liuwei Dihuang series preparations (LDSP). Methods: The network pharmacology technology was used to screen out LDSP, such as pills (big honey pills and concentrated pills), capsules (capsules and soft capsules), granules and oral liquids for the treatment of type 2 diabetes, hypertension and nephrotic syndrome of the common core active ingredients, and the amount of 13 active ingredients (quercetin, kaempferol, β-sitosterol, alisol B, alisol B23-acetate, taxifolin, hydroxygenkwanin, stigmasterol, cerevisterol, hederagenin, catechin, diosgenin and ethyl linolenate) with higher degrees was used as an index of investigation, the HPLC method was used to simultaneously measure the wavelength by switching, the determination was performed on Waters 2695 Supersil C18 (250 mm × 4.6 mm, 5 μm) column with mobile phase consisted of methanol-0.08% phosphoric acid (gradient elution), volume flow rate was 1.0 mL/min, column temperature was 30 ℃, sample size was 10 μL, detection wavelength: first measurement was at full wavelength 200—400 nm, then maximum absorption was switched to detect wavelength according to the reference wavelength; The principal components analysis (PCA) and cluster analysis (CA) method were used to comprehensively evaluate and compare the quality differences of LDSP. Results: Under the same crude drug amount, the content of ingredients in LDSP was significantly different (P < 0.05), and some ingredients were missing in some preparations. Quercetin and catechin were not detected in soft capsules, β-sitosterol were detected only in big honey pills and granules. In the correlation analysis results, the similarity between LDSP was 0.170—0.897, and most of them were concentrated in 0.500—0.600. Big honey pills had the highest similarity with granules, concentrated pills and capsules, soft capsules and the oral solution showed a very significant correlation (P < 0.01), and was significantly related to big honey pills and granules (P < 0.05); The PCA and CA results of LDSP were classified into two categories, big honey pills and granules were classified into one category, and concentrated pills, capsules, soft capsules, and oral liquids were classified into one category. Conclusion: There are certain inherent quality differences in LDSP, which reveals that the phenomenon of completely consistent functional indications in the instructions of LDSP may not be reasonable.

Mechanism of Shaoyao Decoction in treatment of ulcerative colitis based on network pharmacology and molecular docking technology / 中草药

Objective: Network pharmacology and molecular docking technology were used to study the material basis and possible mechanism of Shaoyao Decoction in treatment of ulcerative colitis (UC). Methods: TCMSP and TCMID were used to obtain the potential active components and drug targets of Shaoyao Decoction. GeneCard and OMIM were used to search disease targets. The common targets obtained by matching drug targets and disease targets were imported into String to construct a PPI network, and Cyto NCA plug-in was used to screen key targets. The network diagram was drawn by connecting the key targets with the corresponding components, so as to screen the key components. GO and KEGG enrichment analyses were performed on key targets. SYBYL-X2.0 software was used to dock the molecules of the key components with the key targets. The rat UC model was replicated in vivo. After the intervention of Shaoyao Decoction, the disease activity index was observed, the colonic pathological damage was evaluated, and the levels of TNF-α, IL-4 and CXCR4 were detected by ELISA. Results: A total of 424 potential active components were found in Shaoyao Decoction. The key components included quercetin, palmitic acid, catechin, and procyanidins, etc. Its 41 key targets for UC were mainly related to the positive regulation of transcription, the negative regulation of apoptosis process, the signal transduction, and other biological processes. The key targets played a role in treating UC through signaling pathways such as TNF, HIF-1, cancer pathway, TLR, PI3K-Akt, et al. Molecular docking results showed that key components had good binding activity with corresponding targets. Shaoyao Decoction improved colon pathological damage, down-regulated the level of TNF-α and CXCR4, and up-regulated the level of IL-4 in vivo. Conclusion: Shaoyao Decoction has the characteristics of "multi-components, multi-targets, multi-pathways" in treatment of UC, which lays a foundation for further study of its mechanism.

Selective phytochemicals targeting pancreatic stellate cells as new anti-fibrotic agents for chronic pancreatitis and pancreatic cancer

Activated pancreatic stellate cells (PSCs) have been widely accepted as a key precursor of excessive pancreatic fibrosis, which is a crucial hallmark of chronic pancreatitis (CP) and its formidable associated disease, pancreatic cancer (PC). Hence, anti-fibrotic therapy has been identified as a novel therapeutic strategy for treating CP and PC by targeting PSCs. Most of the anti-fibrotic agents have been limited to phase I/II clinical trials involving vitamin analogs, which are abundant in medicinal plants and have proved to be promising for clinical application. The use of phytomedicines, as new anti-fibrotic agents, has been applied to a variety of complementary and alternative approaches. The aim of this review was to present a focused update on the selective new potential anti-fibrotic agents, including curcumin, resveratrol, rhein, emodin, green tea catechin derivatives, metformin, eruberin A, and ellagic acid, in combating PSC in CP and PC models. It aimed to describe the mechanism(s) of the phytochemicals used, either alone or in combination, and the associated molecular targets. Most of them were tested in PC models with similar mechanism of actions, and curcumin was tested intensively. Future research may explore the issues of bioavailability, drug design, and nano-formulation, in order to achieve successful clinical outcomes with promising activity and tolerability.

Effects of regular green tea intake on body fat and arterial stiffness in young adults: A randomized intervention study / 体力科学

Green tea catechins are well known to be one of polyphenols, and its regular ingestion induces body fat reduction in obese individuals. Cocoa polyphenols of high-cocoa chocolate can also improve arterial stiffness. However, it is unclear whether green tea catechins improve body fat and arterial stiffness even in healthy young adults. Therefore, this study aimed to examine the effects of regular green tea intake on body fat and arterial stiffness in young adults. This randomized, controlled, parallel-group intervention study included 53 Japanese college students (mean age, 21.1 ± 0.1; men, n = 40; women, n = 13). They were randomly divided into three groups: control group (n = 14), intervention I group (n = 19), and intervention II group (n = 20). The participants ingested 500 ml/day of commercially available natural water (0 mg of catechin/day) or green tea (intervention I group, 200 mg of catechin/day; intervention II group, 400 mg of catechin/day) for 4 weeks. After 4 weeks of intervention period, no significant changes in body weight, body mass index, pulse wave velocity, and cardio-ankle vascular index were observed in all groups. However, body fat and carbon dioxide output significantly reduced only in the intervention II group. With the level of catechin concentrations increasing, significant decreasing trends were found in body fat changes and respiratory exchange ratio changes. Therefore, these findings suggest that four weeks of regular green tea intake would reduce body fat, but not arterial stiffness, in young adults.

In vitro tissue culture, preliminar phytochemical analysis, and antibacterial activity of Psittacanthus linearis (Killip) J.K. Macbride (Loranthaceae) / Cultivo de tejidos in vitro, análisis fitoquímico preliminar y actividad antibacteriana de Psittacanthus linearis (Killip) J.K. Macbride (Loranthaceae)

ABSTRACT Hemiparasitic plants commonly known as mistletoe (muérdago in Spanish) in the families Santalaceae and Loranthaceae are common in various kinds of plants or trees, and many hemiparasitic plants are used for medicinal purposes in various parts of the world. The objective of the present work, carried out in Psittacanthus linearis (suelda con suelda), a representative species in the seasonally dry forest (SDF) from the north of Perú, was to study aspects of in vitro tissue culture, carry out preliminary phytochemical analysis, and assess antibacterial activity. Seeds of individuals of P. linearis, which used Prosopis pallida (algarrobo) as host plant, were collected and used to induce in vitro seed germination, clonal propagation, callus induction and organogenesis. Stems, leaves and fruits of individuals of P. linearis were dried, powdered, and subjected to ethanol extraction. Posteriorly the extract was first recovered with ethanol and the remnant with chloroform, which formed the ethanolic and chloroformic fraction. A preliminary phytochemical screening was performed and preliminary antibacterial studies with Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were carried out and their results are discussed. This is the first report about in vitro tissue culture, phytochemical analysis and antibacterial activity of P. linearis. The results may have important implications for understanding physiological and biochemical interactions between host and hemiparasitic species as well as P. linearis with P. pallida and other SDF species.

RESUMEN Las plantas hemiparásitas o 'mistletoe' o 'muérdago' son comunes en varios grupos vegetales o árboles, perteneciendo a las familias Santalaceae and Loranthaceae y muchas plantas hemiparásitas son usadas como medicina en varios lugares del mundo. El objetivo del presente trabajo realizado en Psittacanthus linearis or 'suelda con suelda', especie representativa en el bosque estacionalmente seco (BES) del norte del Perú, fue estudiar algunos aspectos en el cultivo de tejidos in vitro, el análisis fitoquímico preliminar y su actividad antibacterial. Semillas de P. linearis teniendo a Prosopis pallida 'algarrobo' como hospedero, fueron colectadas y utilizadas en la germinación in vitro, propagación clonal, inducción de callos y procesos organogénicos. Tallos, hojas y frutos de plantas silvestres fueron secados, pulverizados y sometidos a extracción con etanol y el extracto fue recuperado primero con etanol y el remanente con cloroformo formando las fraciones etanólica y clorofórmica. Se realizó un estudio fitoquímico y antibacteriano preliminar utilizando Staphylococcus aureus, Escherichia coli y Pseudomonas aeruginosa y los resultados son discutidos. Este trabajo es el primer estudio sobre cultivo de tejidos, análisis fitoquímico y actividad antibacteriana de P. linearis. Los resultados obtenidos tienen importantes implicancias para el conocimiento de las interacciones fisiológicas y bioquímicas entre las especies hospederas y las plantas hemiparásitas, como P. linearis con P. pallida y otras especies del BES.

Influence of green tea extract with different concentrations of epigallocatechin gallate on calvaria bone repair of ovariectomized rats / Influencia del extracto de té verde con diferentes concentraciones de galato de epigalocatequina sobre la reparación ósea de calvaria de ratas ovariectomizadas

Impairing osteoporosis progression is a challenge, and recently the role of antioxidants has been associated to bone metabolism. Green tea extract is rich in catechins, especially epigallocatechin gallate (EGCG), which may help control osteoporosis damage in bone tissue. This investigation evaluated the efficacy of green tea ingestion containing different concentrations of EGCG in calvaria bone repair of ovariectomized rats. Wistar rats (n=15) were ovariectomized and divided into 3 groups: ovariectomized (OVX), ovariectomized + GTE 15 % EGCG (OVX/GTE15), and ovariectomized + GTE 94 % EGCG (OVX/GTE94). Green tea extract was administered by gavage in the concentration of 50 mg/kg and sham group (n=5) received water. Bone defects were performed in the calvaria 60 days after ovariectomy followed by 4 weeks until euthanasia. Bone samples were collected to perform qualitative and quantitative histological analysis of bone formation. Data obtained were submitted to normality and ANOVA statistical test for p<0.05. The mean values of neoformed bone for Sham, OVX, OVX/GTE15 and OVX/GTE94 were respectively: 21.11 ± 3.91; 19.92 ± 2.20; 33.05 ± 1.26 e 34.75 ± 0.54 (p<0.05). Results show that continuous ingestion of green tea extract immediately after ovariectomy shows positive effects in the prevention of bone loss in osteoporosis, even with low concentrations of EGCG.

La disminución en la progresión de la osteoporosis es un desafío, y recientemente el papel de los antioxidantes se ha asociado al metabolismo óseo. El extracto de té verde es rico en catequinas, especialmente el galato de epigalocatequina (EGCG), lo que puede ayudar a controlar el daño de la osteoporosis en el tejido óseo. Esta investigación evaluó la eficacia de la ingesta de té verde con diferentes concentraciones de EGCG en la reparación ósea de calvaria de ratas ovariectomizadas. Las ratas Wistar (n = 15) fueron ovariectomizadas y divididas en 3 grupos: ovariectomizadas (OVX), ovariectomizadas + GTE 15 % EGCG (OVX / GTE15), y ovariectomizadas + GTE 94 % EGCG (OVX / GTE94). El extracto de té verde se administró por sonda en una concentración de 50 mg/kg y el grupo simulado (n = 5) recibió agua. Los defectos óseos se realizaron en la calvaria 60 días después de la ovariectomía, seguido de 4 semanas hasta la eutanasia. Se obtuvieron muestras de hueso para realizar un análisis histológico cualitativo y cuantitativo de la formación ósea. Los datos obtenidos se sometieron a normalidad y prueba estadística ANOVA (p<0,05). Los valores medios de hueso neoformado para Sham, OVX, OVX / GTE15 y OVX / GTE94 fueron: 21,11 ± 3,91; 19,92 ± 2,20; 33,05 ± 1,26 y 34,75 ± 0,54 (p <0,05), respectivamente. Los resultados muestran que la ingesta continua de extracto de té verde, inmediatamente después de la ovariectomía, muestra efectos positivos en la prevención de la pérdida ósea ocurrida en la osteoporosis, incluso con concentraciones bajas de EGCG.

Mechanical-physicochemical properties and biocompatibility of catechin-incorporated adhesive resins

Abstract Several anti-proteolytic dentin therapies are being exhaustively studied in an attempt to reduce dentin bond degradation and improve clinical performance and longevity of adhesive restorations. Objectives This study assessed the effect of epigallocatechin-3-gallate (EGCG) on long-term bond strength when incorporated into adhesives. Material and Methods Adhesive systems were formulated with EGCG concentrations of 0 wt%: (no EGCG; control); 0.5 wt% EGCG; 1.0 wt% EGCG, and 1.5 wt% EGCG. Flexural strength (FS), modulus of elasticity (ME), modulus of resilience (MR), compressive strength (CS), degree of conversion (DC), polymerization shrinkage (PS), percentage of water sorption (%WS), percentage of water solubility (%WL) and cytotoxicity properties were tested. Dentin microtensile bond strength (µTBS) was evaluated after 24 h and again after 6 months of water storage. The adhesive interface was analyzed using scanning electron microscopy (SEM). Results No significant differences were found among the groups in terms of FS, ME, MR, CS and PS. EGCG-doped adhesives increased the DC relative to the control group. EGCG concentrations of 1.0 wt% and 0.5 wt% decreased the WS of adhesives. WL decreased in all cases in which EGCG was added to adhesives, regardless of the concentration. EGCG concentrations of 1.0 wt% and 0.5 wt% reduced cytotoxicity. EGCG concentrations of 1.0 wt% and 0.5 wt% preserved µTBS after 6 months of storage, while 1.5 wt% EGCG significantly decreased µTBS. SEM: the integrity of the hybrid layer was maintained in the 0.5 wt% and 1.0 wt% EGCG groups. Conclusion EGCG concentrations of 1.0 wt% and 0.5 wt% showed better biological and mechanical performance, preserved bond strength and adhesive interface, and reduced cytotoxicity.

A new rhamnoside from Aspidopterys obcordate / 中草药

Objective To study the chemical constituents of Aspidopterys obcordate. Methods The chemical constituents from 95% EtOH extract of the stem of A. obcordate were isolated by repeated chromatograph with silica gel, Sephadex LH-20, and ODS. The structures were elucidated by spectroscopic analysis. Results Eight compounds were isolated and identified as 3,4-dihydroxybenzoic acid-3-O-α-L-rhamnoside (1), 3,4-dihydroxyphenyl acetate (2), 3,4-dihydroxybenzoic acid (3), catechin (4), β-sitosterol (5), daucosterol (6), cinnacasolide C (7), and alantan (8). Conclusion Compound 1 is a new compound, named as apobcoroside. All compounds are isolated from the plant of A. obcordate for the first time.

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.

Determination of 25 characteristic components in Danhe Granules by LC-MS and consistency analysis of preparations / 中草药

Objective: A high performance liquid chromatography coupled to triple quadrupole mass spectrometry method (HPLC- MS/MS) was established to simultaneously determine the content of 25 characteristic components (gallic acid, tanshinol, catechin, chlorogenic acid, caffeic acid, epicatechin, rutin, polydatin, hyperin, astragalin, naringin, hesperidin, rosmarinic acid, resveratrol, salvianolic acid B, quercetin, emodin-8-O-β-D-glucoside, isorhamnetin, emodin, coclaurine, nuciferine, cryptotanshinone, tanshinone I, dehydronuciferine, tanshinone IIA) in Danhe Granules (DG), and the consistency between different batches was investigated. Methods The analysis was conducted on Agilent Rapid Resolution HD C18 column (50 mm × 2.1 mm, 1.8 μm) with the mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile. The dynamic multi-response detection (dMRM) scanning mode was used in the mass spectrometry. Results: Based on the established HPLC-MS/MS method, the simultaneous quantitative analysis of 25 characteristic components could be completed within 10 min, with the quantitative limits of isorhamnetin, coclaurine, and dehydronuciferine of 0.025 ng/mL; emodin and tanshinone I of 0.050 ng/mL; emodin-8-O-β-D-glucoside of 0.200 ng/ml; gallic acid, chlorogenic acid, caffeic acid, epicatechin, rutin, hyperin, astragalin, resveratrol, quercetin, nuciferine, tanshinone IIA of 0.250 ng/ml; catechin, rosmarinic acid, and cryptotanshinone of 0.500 ng/mL; tanshinol, hesperidin, and salvianolic acid B of 1.000 ng/mL; polydatin of 2.000 ng/mL; naringin of 5.000 ng/mL, respectively. The linear relationships of the 25 constituents within their respective mass concentrations were good, with the average recovery of 85.16%-113.46% and the RSD of 2.01%-8.80%. Furthermore, this method also included the main components named monarch, minister, assistant and guide of herbs in a relatively comprehensive way. The total content of the 25 components was 31.49 mg/g, among which the content of salvianolic acid B (9.44 mg/g) and hesperidin (7.60 mg/g) was the highest, and the content of isorhamnetin (0.79 μg/g) was the lowest. According to boxplot analysis, the content of 25 components in 10 different batches of DG fluctuated (P value) within 75% < P < 125%; and the RSD value of 25 components ranged from 2.58% to 13.10% by statistical analysis. The above results showed that the consistency of the component content among 10 batches of DG was acceptable. Conclusion: The analytical method established in this study is fast and sensitive. Furthermore, the results of this study are reliable and can provide scientific methods and basis for quality control and consistency analysis of DG.

Simultaneous Determination of Gallic Acid ，Cinnamic Acid and Catechin in 3 Processed Products of Rheum officinale by RP- HPLC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of gallic acid， cinnamic acid and catechin in 3 processed products of Rheum officinale. METHODS： RP-HPLC method was established. The determination was performed on Thermo ScientificTM Hypersil GOLD Dim column with mobile phase consisted of methanol-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 278 nm， and the column temperature was 30 ℃. The sample size was 10 μL. RESULTS： The linear range of gallic acid， cinnamic acid and catechin were 0.126 2-1.262 0 μg（r＝0.999 9）， 0.036 2-0.362 0  μg（r＝0.999 9） and 0.177 9-1.779 4 μg（r＝0.999 8）， respectively. Quantitative limits were 25.4， 28.2， 62.5 ng， and detection limits were 6.2， 3.6， 11.8 ng， respectively. RSDs of precision， stability， repeatability and durability tests were all less than 3％. The recoveries ranged from 94.64％-102.71％（RSD＝2.74％， n＝9）， 95.35％-102.49％（RSD＝2.44％， n＝9）， 93.56％-103.66％（RSD＝3.27％， n＝9）. The determination results showed that the contents of gallic acid and cinnamic acid in prepared R. officinale were higher， and the order of both were prepared R. officinale＞steamed R. officinale＞raw R. officinale. The content of catechin in raw R. officinale was higher， and the order of it was raw R. officinale＞ steamed R. officinale＞prepared R. officinale. CONCLUSIONS： The method is sensitive， reliable and reproducible. It can be used to determine the contents of gallic acid， cinnamic acid and catechins in 3 processed products of R. officinale simultaneously.

Simultaneous Determination of Contents of 8 Components in Eucommia ulmoides Leaves by HPLC / 中国药房

OBJECTIVE： To establish content determination method for simultaneously determining aucubin，geniposidic acid，catechin，chlorogenic acid，asperuloside，rutin，isoquercitrin and astragalin in Eucommia ulmoides leaves. METHODS：HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with the mobile phase consisted of acetonitrile-0.1％ phosphoric acid（gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for aucubin and catechin，239 nm for geniposidic acid and asperuloside，220 nm for chlorogenic acid，354 nm for rutin and isoquercitrin，and 266 nm for astragalin. The sample size was 5 μL. RESULTS： The linear range of aucubin， geniposidic acid， catechin， chlorogenic acid， asperuloside， rutin， isoquercitrin and astragalin were 0.812-6.090 μg（r＝0.999 3），0.438-3.285 μg（r＝0.999 2），0.045-0.336 μg（r＝0.999 2），0.882-6.615 μg（r＝0.999 3），0.097-0.726 μg（r＝0.999 1），0.064-0.483 μg（r＝0.999 3），0.048-0.360 μg（r＝0.999 1） and 0.014-0.108 μg（r＝0.999 7），respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.5％（n＝6）. The average recovery rates were 101.60％，103.06％，99.77％，96.93％，98.17％，96.75％，98.97％ and 99.60％，with RSDs of 1.42％，2.65％，2.78％，2.05％，2.26％，0.93％，2.79％ and 3.08％，respectively（n＝6）. The contents of aucubin， geniposide， catechin， chlorogenic acid， plantain， rutin， isoquercetin and astragaloside in 12 batches of E. ulmoides leaves from different collection time and planting varieties were 10.903-17.245， 5.578-7.892， 0.198-0.440， 13.890-19.782， 1.008-1.547， 1.102-2.396， 0.267-0.701， 0.150-0.412 mg/g， respectively. The content fluctuated greatly. The contents of aucubin， geniposide， catechin， chlorogenic acid， rutin and pinoresinol diglucoside in cortex of E. ulmoides were 0.299， 0.123， 0.580， 0.112， 0.026，1.961 mg/g， respectively. CONCLUSIONS： The method is simple， reproducible and accurate. It can be used to evaluate the quality of E. ulmoides leaves. There are obvious differences in composition and content of components in different medicinal parts （cortex， leaves） of E. ulmoides.

Simultaneous Determination of 8 Non-anthraquinone Constituents in Rheum palmatum by HPLC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of 8 non-anthraquinone constituents in Rheum palmatum. METHODS： HPLC method was adopted. The determination was performed on Symmetry C18 column with mobile phase consisted of methanol-0.1％ phosphoric acid （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ and detection wavelength was 280 nm. Sample size was 30 μL. RESULTS： The linear range of gallic acid， catechin， epicatechin， resveratrol 4′-O-glucopyranoside， epicatechin gallate， resveratrol 4′-O-β-D-（6″-O-galloyl）-glucopyranoside， sennoside A， 4′-hydroxyphenyl-2-butanone-4′-O-β-D-（2″-O-galloyl-6″-O-p-hydroxy cinnamyl）-glucopyranoside were 6.16-2 464 ng（r＝0.999 9）， 37.4-14 960 ng（r＝0.999 9）， 7.635-3 054 ng（r＝0.999 7）， 7.63-3 052 ng（r＝0.999 9）， 8.32-3 328 ng（r＝0.999 9）， 11.5-4 600 ng（r＝0.999 9）， 16.08-6 432 ng（r＝0.999 9）， 29.3-11 720 ng（r＝0.999 9）， respectively. The limits of quantitation were 3.48， 4.30， 6.40， 4.40， 3.39， 2.87， 8.40 and 4.95 ng， respectively. The limits of detection were 2.32， 2.58， 2.40， 2.64， 2.26， 1.23， 4.20， 2.97 ng， respectively. RSDs of precision， stability and reproducibility tests were all lower than 5％. Recoveries were 94.32％- 100.54％（RSD＝2.78％，n＝6）， 91.15％-99.36％（RSD＝3.72％，n＝6）， 92.16％-98.04％（RSD＝2.39％，n＝6）， 93.41％-100.73％（RSD＝3.17％，n＝6）， 93.89％-98.40％（RSD＝1.99％，n＝6）， 92.61％-101.74％（RSD＝3.71％，n＝6）， 92.66％-103.40％（RSD＝3.76％，n＝6）， 95.45％-102.70％（RSD＝3.06％，n＝6）， respectively. CONCLUSIONS： The established method is simple， accurate and specific， and can be used for the simultaneous determination of 8 non-anthraquinone constituents in R. palmatum.