Imperatorin alleviates chronic endometrial inflammation by inhibiting the NLRP3 inflammatory pathway in rats / 免疫学杂志

This study was designed to explore the treatment effect of imperatorin(Imp)on chronic endometritis(CE)and the possible mechanism.A rat model of CE was established by injecting phenol gel into the ovary and was grouped into CE group,Imp low dose group(ImpL group,5 mg?kg-1?d-1),Imp high dose group(ImppH group,10 mg?kg-1?d-1),and ImpH+lipopolysaccharide(LPS,10 mg/kg)groups,while sham-treated rats were grouped as control group(CT group).Uterine organ index of all groups were measured;hematoxylin eosin(HE)staining was applied to observe the pathological changes of uterine tissue;immunohistochemistry was applied to detect the expression of vascular endothelial growth factor(VEGF)in the endometrium;ELISA was used to measure the levels of serum IL-6 and TNF-α;RT-qPCR and Western blot were applied to verify the expression of IL-1β,nuclear factor-κB(NF-κB),NOD-like receptor protein 3(NLRP3),and matrix metalloproteinase(MMP)-9 in uterine tissue.Compared with the CT group,the uterus and vagina of the CE group were swollen and congested,and the expression of VEGF in the endometrium was decreased(P<0.05),the levels of IL-6 and TNF-α in serum and the expressions of IL-1β,p-NF-κB/NF-κB,NLRP3,and MMP-9 in uterine tissue were increased(P<0.05).Imp could reverse these pathological changes mentioned above in CE model,while LPS was able to partially suppress the improvement effect of ImpH on CE rats.In conclusion,Imp may have a certain therapeutic effect on CE by inhibiting the activation of NLRP3 inflammatory pathway.

Study on improvement effects and mechanism of imperatorin on cachexia model mice / 中国药房

OBJECTIVE To investigate the improvement effects and mechanism of imperatorin on cachexia model mice. METHODS Fifteen male C57BL/6J mice were randomly divided into blank control group， model group and imperatorin group， with 5 mice in each group. Except for blank control group， the remaining mice were inoculated with LLC cell suspension subcutaneously on the dorsal surface， and the drug was administered by gavage daily from the 7th day of inoculation. The imperatorin group was gavaged with imperatorin suspension （0.5% sodium carboxymethylcellulose solution as solvent） at 60 mg/kg； blank control group and model group were given an equal volume of 0.5% sodium carboxymethylcellulose solution， for 13 d of continuous administration. During the administration period， food intake and body mass of mice were recorded daily and regularly， tumor long and short diameters were measured every two days， and tumor volume was calculated. The skeletal muscle mass and tumor mass of each group were weighed and the tumor-free body weight was calculated； the pathological changes of skeletal muscle were observed and the cross-sectional area of skeletal muscle fibers was calculated； the phosphorylation levels of signal transduction and activator of transcription 3 （STAT3） （measured as p-STAT3/STAT3 ratio）， muscle atrophy box F gene （MAFbx）， myostatin （Myog）， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， Caspase3 protein and mRNA expression were all detected. RESULTS Compared with blank control group， body mass and skeletal muscle mass of model group were decreased significantly （P＜0.05）， and reduced food intake， loose arrangement of skeletal muscle， large cell space were observed； the cross-sectional area of skeletal muscle fiber was significantly reduced， while p-STAT3/STAT3 ratio， protein and mRNA expressions of MAFbx， Bax and Caspase3 were somo_amour@163.com increased significantly （P＜0.05）. The protein and mRNA expressions of Myog and Bcl-2 were significantly reduced （P＜ 0.05）. Compared with model group， body weight， tumor-free weight and skeletal muscle weight were increased significantly in imperatorin group （P＜0.05）； food intake increased， while the expressions of tumor weight and volume were decreased significantly （P＜0.05）； the expressions of above proteins and genes were improved significantly （P＜0.05）. CONCLUSIONS Imperatorin can improve the tumor cachexia state， the mechanism of which may be related to the regulation of ubiquitin-proteasome pathway and anti-apoptosis.

Mechanisms of imperatorin on regulating P-glycoprotein in blood-brain barrier based on network pharmacology and in vitro experi-ment / 中国药理学与毒理学杂志

OBJECTIVE To explore mecha-nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net-work pharmacology combined with in vitro experi-ment.METHODS Drug targets were predicted using the Pharmapper and Swiss targets data-bases;disease targets were obtained through the Genecards database;intersections between drugs and disease targets were screened by Cytoscape software;the obtained core targets were used to construct protein-protein interaction(PPI)network,gene ontology(GO)functions,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The effects of imperatorin(20,50,100 μ mol·L-1)on P-gp activity were monitored in hCMEC/D3in vitro BBB model,and the effects of imperatorin on the expression of target proteins were verified using Western blot method.RESULTS 55 drug targets and 3102 disease targets were obtained from the network pharmacology screening,and 37 core targets were obtained after the combination.Enrichment analysis showed that core targets were closely related to chemical synaptic trans-mission regulation,neurotransmitter receptor activity,proteinkinaseregulationactivity,G protein-coupled receptor signaling pathway,neural active ligand receptor interaction pathway,PI3K-Akt sig-naling pathway,VEGF signaling pathway,etc..In vitro experimental validation suggested that all tested concentration groups of imperatorin signifi-cantly reduced the activity and expression of P-gp,which were achieved by significantly downregu-lating the phosphorylation levels of PI3K and Akt,and repressing the expression of VEGFR2 pro-tein.CONCLUSION Network pharmacology was used to predict the core targets and signaling pathways of imperatorin on regulating P-gp in BBB and relevant validation was conducted through in vitro experiments,providing a refer-ence basis for further exploration of the mecha-nisms of imperatorin on regulating P-gp in BBB.

Effect of Angelica dahurica extract and imperatorin on gouty arthritis / 中国药理学通报

Aim To investigate the effects of Angelica dahurica extract and imperatorin on gouty arthritis induced by monosodium urate(MSU)crystal.Methods The model of gouty arthritis was established by injecting MSU crystals into the left ankle of rats.The degree of ankle swelling was measured at 0-24 h in rats.The histopathological changes of synovial tissues were observed.Meanwhile the mRNA and protein expression levels of Nod-like receptor protein 3(NLRP3), interleukin-1β(IL-1β), cysteinyl aspartate specific proteinase-1(caspase-1), interleukin-1 receptor type 1(IL-1R1), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)were detected by Western blot and RT-PCR in synovial tissues.Results Angelica dahurica extract and imperatorin significantly reduced MSU induced left ankle swelling in rats, and improved inflammatory infiltration in synovial tissues.The expression of NLRP3, IL-1β, caspase-1, IL-1R1, IL-6, TNF-α protein or mRNA in synovial tissues decreased significantly.Conclusion Angelica dahurica extract and its active component imperatorin could ameliorate gouty arthritis, which supplys basic data support for its clinical application.

Study on HPLC fingerprints of Biyuanjing capsules / 药学实践杂志

Objective To establish a HPLC fingerprints of Biyuanjing capsules. Methods The column was Agilent SB-C18(4.6mm×250 mm, 5 µm). The mobile phase was acetonitrile-water with gradient elution at a flow rate of 1.0 ml/min. The detection wavelength was 210 nm. The detection time was 80 min. Results The HPLC fingerprints of Biyuanjing capsules were established. Twenty common peaks were confirmed, of which, 15 peaks were belonging to each crude drug and 5 peaks were identified as chemical components. The overall similarity of the fingerprints of 10 batches of samples was above 90% comparing with the control. Conclusion This method can be used for the quality control of Biyuanjing capsules.

Research Progress on Pharmacological Effect and Mechanism of Imperatorin / 中国实验方剂学杂志

Imperatorin, which is extracted from umbelliferous plants such as Radix Angelicae Dahuricae, Radix Saposhnikoviae and Fructus Cnidii, belongs to furanocoumarins and is especially rich in Radix Angelicae Dahuricae. Research has shown that imperatorin possesses functions of anti-inflammatory, analgesic, antibactrial, antiviral, antiallergic, anti-tumor, reverse drug resistance in tumor cells, interaction with drug metabolizing enzymes, affecting cardiovascular and nervous system effect. It is also one of the standard components in quality control of various analgesics. In recent years, research findings related to imperatorin is increasing fast. A number of patent applications have been approved for the application of imperatorin in the treatment of anti-tumor and cardiovascular diseases. In addition, since the water insolubility of imperatorin affects its bioavailability, most researchers have gradually attached importance to this aspect of research, such as modifying its structure or synthesizing its derivatives. The literatures on the pharmacological effects and mechanisms of imperatorin at home and abroad in recent years were consulted and summarized in this paper. Imperatorin was found not only to display other pharmacological effects like furanocoumarins but also could cure osteoporosis, skin diseases and show photosensitization. Moreover, the mechanism of its action has the effect of multi-pathway and multi-target, but most of the studies have not identified its targets, which still needs further study. Extensive and significant pharmacological effect make imperatorin show a great potential for development of new drugs. This paper reviews the basic properties, the progress on pharmacological effects and mechanisms of imperatorin, proposes the research status and direction of future reseach. Hopes to provide ideas for researchers and beneficial references for the future development and utilization of imperatorin.

Chemical constituents of Artemisia rupestris / 中草药

Objective: To investigate the chemical constituents of the whole plants of Artemisia rupestris. Methods: The chemical constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography and semi-preparative HPLC. The structures of the isolated compounds were identified by NMR and MS spectroscopic method. Results: Eighteen compounds were isolated and purified, which structures were identified as 2α-(2',4'-hexadiynoyl)-1,6-dioxaspiro [4,5]-deca-3-ene (1), chrysophanol (2), ethyl vanillate (3), methyl linolenate (4), 8,11-octadecadienoic acid methyl ester (5), eicosane coumaric acid ester (6), emodin (7), 2,5-di-tert-butylphenol (8), phellopterin (9), syringic acid (10), 7-methoxycoumarin (11), vanillin (12), imperatorin (13), oroxylin A (14), 5-hydroxy-3,4',6,7-tetramethoxyflavone (15), erucylamide (16), octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate (17) and 3,5,3',4'-tetrahydroxy-6,7-dimethoxyflavone (18). Conclusion: Compounds 1, 4, 11-14 and 18 are isolated from A. rupestris for the first time, and all the other compounds were isolated from the genus Artemisia for the first time.

Study on coumarins and alkaloids from stems of Clausena lenis / 中草药

Objective: To study the coumarins and alkaloids from the stems of Clausena lenis. Methods: The chemical constituents from the stems of C. lenis were separated and purified by silica gel, ODS, Sephadex LH-20 gel column chromatographies and preparative HPLC. Their structures were identified by physicochemical properties, spectroscopic analysis, as well as the comparisons with the data reported in literatures. Results: A total of 18 compounds were isolated from the 90% ethanol extract of the stems of C. lenis, which were identified as 6,8-diprenylumbelliferone (1), byakangelicin (2), tert-O-methylheraclenol (3), pabularinone (4), isogosferol (5), heraclenol (6), imperatorin (7), decursinol (8), xanthyletin (9), marmesin (10), skimmianine (11), kokusaginine (12), N-metilatanina (13), clausine Z (14), murrayanine (15), 2-methoxy-1-(3-methyl-buten-1-yl)-9H-carbazole-3-carbaldehyde (16), claulansine I (17) and clausehainanine A (18). Among them, compounds 1-10 were coumarins and 11-18 were alkaloids. Conclusion: All compounds are isolated from C. lenis for the first time, compounds 2, 3 and 8 are separated from the genus Clausena for the first time.

HPLC fingerprint of Cnidii Fructus formula granule and determination of six main coumarin components / 中草药

Objective: To establish an HPLC fingerprint of Cnidii Fructus formula granule analysis method for simultaneous determination of six main coumarin components, including osthol, xanthotoxin, xanthotol, bergapten, imperatorin and isopimpinellin, in order to provide reference for the study of the material basis of Cnidii Fructus formula granule. Methods: The method was performed by high performance liquid chromatography with a Waters XBridge C18 (250 mm × 4.6 mm, 5 μm) column and methanol (A)-0.1% acetic acid (B) as the mobile phase for gradient elution. The flow rate was 0.5 mL/min, the injection volume was 10 μL and the column temperature was 40 ℃. The detection wavelength was set at 320 nm. The chromatographic fingerprint evaluation system published by the State Pharmacopoeia Commission (2012 Edition) was used to establish the fingerprint of Cnidii Fructus formula granule, and the content of six main coumarin components was simultaneously determined. Results: The research on the 18 batches of Cnidii Fructus formula granule showed that the fingerprint similarity was greater than 0.992 and 19 common peaks were calibrated with satisfied peak resolution. The content determination results showed that the content of both xanthotoxin and osthol were the main coumarin components in Cnidii Fructus formula granule. According to the methodological investigation, the precision RSD values were all less than 1.6%. The sample was stable within 48 h and this method had good repeatability. The average recovery rates of xanthotol, xanthotoxin, imperatorin, isopimpinellin, bergapten and osthol were 100.69%, 101.03%, 99.48%, 100.88%, 101.27% and 100.35%, respectively. All of these coumarin components’ RSD were less than 2.5%. The six components showed a good linear relationship within a certain concentration range. The results of the content determination of xanthotol, xanthotoxin, isopimpinellin, bergapten, imperatorin and osthol respectively were 8.01-8.29, 2.37-2.63, 4.30-4.61, 4.04-4.40, 3.45-3.90 and 6.02-6.80 mg/g among the 18 batches of the Cnidii Fructus formula granule. Conclusion: The fingerprint method and the determination method of six main coumarin components in the Cnidii Fructus formula granule established in this study are simple, stable, accurate and reliable. This method can be used for the quality control of the Cnidii Fructus formula granule.

Preparation and Quality Evaluation of Imperatorin Ultradeformable Liposomes Gel / 中国药房

OBJECTIVE：To prepare Imperatorin ultradeformable liposomes gel （IMP-UDLs-Gel），and to evaluate its quality. METHODS：Based on single factor test ，using 12 h accumulative penetration amount （Q12h）as evaluation index ，the proportion of carbomer 940，glycerol and propyl glycol in formulation of IMP-UDLs-Gel were investigated by orthogonal test. The optimal formulation was screened. The quality of IMP-UDLs-Gel prepared with the optimal formulation was evaluated. RESULTS ：The optimal formulation of IMP-UDLs-Gel included carbomer 940 proportion of 1％，glycerol proportion of 15％ and propyl glycol proportion of 10％. Q12 h of IMP-UDLs-Gel was （11.543±0.241）μg/cm2；the appearance was milky white and translucent ；the particle size was （93.13±1.68）nm，PDI was 0.268±0.012，Zeta potential was （－24.96±1.99）mV；pH was 7.32±0.03； viscosity was （45.37±1.27）g·s；steady flow was （0.727±0.002）μg·h/cm2，lag time was （4.358±0.175）h，apparent permeability coefficient was 1.392×10－3 cm/h，and it has good physical and optical stability. CONCLUSIONS ：The preparation method is stable and feasible ，and the prepared IMP-UDLs-Gel has good adhesion ，stability and transdermal property.

Antidepressant effect and mechanism of imperatorin / 中国药理学通报

Aim To investigate the antidepressant effect of imperatorin(IMP) and explore its mechanism. Methods The model of depression was established by prenatal stress (PS) on late pregnant mother, then male offspring rats were randomly divided into five groups: control, PS model, fluoxetine group(5 mg • kg"1), IMP( 15, 30 mg • kg"1). Sucrose preference test, forced swimming test, and open field test were used to evaluate the behavioral changes. 5- hydroxytryptamine ( 5-HT ) concentration, serotonin transporters(5-HTT) and 5-HT,A receptor(5-HT,AR) mRNA expression in the hippocampus and frontal cortex were measured using an enzyme-linked immunosorbent assay and real-time PCR. Results Compared with PS group, the percentage of sucrose preference and the number of total crossings, center crossings, rearing, grooming and 5-IIT concentration significantly increased in IMP group(/> <0. 01). The im-mobility time was significantly reduced in IMP group compared with PS group ( P < 0. 01). The 5-HT concentration and 5-HT1A R mRNA expressions of PS offspring rats significantly increased by the administration ofIMP (P<0.05,P<0.01). And, the increased 5- HTT mRNA markedly decreased in IMP group compared with PS group (P < 0. 05 , P < 0. 01 ). Conclusions IMP can obviously improve rat behavior and show anti-depressant effect, which may relate to the 5- HT concentration, 5-HTT and 5-HT,AR mRNA expression in hippocampus and prefrontal cortex.

Chemical constituents and analgesic activity of the aerial parts of skimmia multinervia / 中国药学杂志

OBJECTIVE: To study the chemical constituents and analgesic activity of the aerial parts of Skimmia multinervia. METHODS: The chemical constituents were isolated by column chromatography packed with MCI and silica gel, respectively, and the structures of the purified compounds were elucidated by NMR and MS analysis. The analgesic activities of the extract, fractions and selected compounds from the aerial parts of S. multinervia were evaluated by using acetic acid-induced writhing test in mice model. RESULTS: The structures of compounds 1-13 are identified as skimmin (1), imperatorin (2), auraptene (3), umbelliferon (4), scopoletin (5), limonin (6), limonexic acid (7), nomilin (8), citrusin (9), taraxerol (10), kanalpin (11), skimmianine (12), and methyl vanillate (13). The extract, ethyl acetate fraction, and alkaloid fraction could reduce the times of mice writhing induced by acetic acid, and the analgesic effect of compound 3, with inhibition rate of 88.18%, was much better than that of the positive control, aspirin. CONCLUSION: Compounds 1-13 are isolated from the plant for the first time, while 7, 9, 11, and 13 are firstly isolated from Skimmia genus. The analgesic activities of S. multinervia and compound 3 are reported for the first time.

Analysis of nationwide random inspection results of Angelicae dahurica and establishment of a quantitative analysis method of multi-component with a single-maker / 中草药

Objective: To analyze the quality of Angelicae Dahuricae Radix (ADR) from national drug evaluation sampling, continuously improve and revise the standard determination method, and ensure the safety of public drug use, six index components of ADR medicinal material and decoction pieces were determined by combining HPLC fingerprint chromatography and quantitative analysis of multi-components by single marker (QAMS). Methods: The determination and exploratory study were carried out according to the National Standard. The establishment of steamed Angelicae dahurica HPLC fingerprint, method for multiple assessment of six index components to conduct a methodological survey determination of multi-index component content of 20 batches of Angelicae dahurica tablets, the final determination of the quality of Angelicae dahurica. Result:s A total of 239 batches of ADR were received from 189 enterprises, in which 225 batches were qualified and 14 batches were unqualified. The qualified rate was 94%. A method that determined content of six kinds of coumarin in ADR decoction pieces using imperatorin as control was established, which could solve the problem of difficulty in obtaining control and consuming long determination time. It would better indicate the drug quality. Conclusio:n The method we had established could be used in qualitative and quantitative analysis of oxypeucedanin hydrate, byakangelicin, oxypeucedanin, berapten, imperatorin and isoimperatorin in ADR decoction pieces. Through this random inspection, it is found that there were still some quality problems in ADR decoction pieces. The quality supervision of traditional Chinese medicine should be strengthened further.

Optimization of extraction process of Xiaoruzeng Capsules based on orthogonal design in information entropy theory / 中草药

Objective: To optimize the extraction process parameters of seven herbs in Xiaoruzeng Capsules (XC). Methods: The content of paeoniflorin, ferulic acid, hesperidin, imperatorin and ligustilide was determined by HPLC. The extraction rate of five kinds of index components, the dry extract yield of extracted herbs and the fingerprint similarity of extracts were comprehensively evaluated. The orthogonal experiment was carried out to investigate the effects of ethanol concentration, extraction solvent, extraction time and extraction times on the extraction process. The information entropy weighting method was used to determine the objective weight of each index, and the extraction process parameters of seven herbs in XC were optimized. Results: According to the comprehensive scoring results, it was determined that the best extraction process of the preparation was to add six times the amount of 50% ethanol, and decocted two times for 2 h each time. The average score of the three batches of verification scores was 99.72, and the RSD was 0.24%. Conclusion: The preferred process has high extraction rate, with good stability and repeatability, which is suitable for mass production of the preparation.

Fingerprinting study on Niuhuang Shangqing Pills based on comprehensive two-dimensional liquid chromatography / 中草药

Objective: To develop a new three-dimensional fingerprinting method and its assessing Methods: based on comprehensive two-dimensional liquid chromatography using Niuhuang Shangqing Pills (NSP) as an example. The developed method could offer new method for the quality control of NSP. Methods: In first dimension, the separation was achieved with an Acquity UPLC HSS CYANO column (100 mm × 2.1 mm, 1.8 μm), methanol-0.1% formic acid in water were used as mobile phases, flow rates were 0.1 mL/min. In second dimension, the separation was achieved with a Kinetex Phenyl-Hexyl column (50 mm × 3 mm, 2.6 μm), acetonitrile-0.1% formic acid in water were used as mobile phases, flow rates were 1.5 mL/min, detection wavelength was set at 254 nm, and acquiring frequency was at 12.5 Hz. Column temperature for each dimension was 40 ℃ and volume of loop linking the two dimensions was 100 μL. Three similarity-calculating Methods:, Euclidean Distance, Cosine, and Correlation Coefficient, were employed to assess the similarities among the 21 samples on the market using medians with arithmetic means of peak volumes of the common peaks as control fingerprints. Results: The three-dimensional fingerprints of 21 batches of NSP samples on the market were developed; Eighteen common peaks were assigned and five of them were identified, which were geniposide (1), pulegone (8), baicalin (9), imperatorin (15), and wogonin (16). Conclusion: A three-dimensional fingerprinting method and its assessing Methods: based on comprehensive two-dimensional liquid chromatography using NSP as an example were successfully developed for the first time, suggesting that it is a feasible method for developing fingerprints for Chinese materia medica. This work improves and supplements the traditional liquid-chromatography fingerprints.

Content Determination of Four Index Components in Compound Baizhi Zhitong Capsules by QAMS Method / 中国中医药信息杂志

Objective To develop a new method for content determination of chlorogenic acid, ferulic acid, and isoimperatorin in Compound Baizhi Zhitong Capsules through quantitative analysis of multi-components by single-marker (QAMS) method by setting imperatorin as internal standard substance. Methods HPLC was adopted. The chromatographic column of Thermo Hypersil GOLD (4.6 mm×250 mm, 5 μm) was used; the mobile phase was composed of acetonitrile and 0.2％ formic acid in gradient elution; the flow speed was 1 m L/min; the detection wavelength were 320 nm (chlorogenic acid, ferulic acid) and 300 nm (imperatorin and isoimperatorin). The relative analysis correction factors of imperatorin to other three components were established by setting imperatorin as internal standard substance. Results The relative analysis correction factors of chlorogenic acid, ferulic acid, and isoimperatorin in Compound Baizhi Zhitong Capsules were 1.16, 2.00, and 1.00, which had a high reproducibility. There was no significant difference between QAMS and external standard method. Conclusion The QAMS method for Compound Baizhi Zhitong Capsules by setting imperatorin as internal standard substance is accurate and feasible, which can be used as a useful quality control method for Compound Baizhi Zhitong Capsules.

Study on Quality Standard of Cnidium monnieri Liniment / 中国药房

OBJECTIVE:To formulate the quality standard of Cnidium monnieri liniment (called "liniment" for short)preliminarily. METHODS:The property of liniment was observed. TLC was used for qualitative identification of osthole in liniment;relative density,alcohol content and pH value were also determined. The osthole and imperatorin in liniment were analyzed qualitatively by HPLC. RESULTS:3 batch of samples were reddish brown liquid and fragrant smelling. In TLC of test sample,the same color fluorescent spots were found in corresponding position as chromatogram of control sample. The linear range of osthole and imperatorin were 0.101 2-0.910 8(r=0.999 6)and 0.006 2-0.124 4 mg/L(r=0.999 6). RSDs of precision tests were 1.38% and 0.79%(n=6). RSDs of stability tests were 0.33% and 0.41%(n=6). RSDs of reproducibility tests were 0.83% and 1.98%(n=6),respectively. Average recoveries rate were 98.73%(RSD=1.29%,n=6)and 99.25%(RSD=1.22%,n=6).Results of content determination showed that the content of osthole and imperatorin were 2.20-2.35 mg/mL and 0.310-0.340 mg/mL,respectively. CONCLUSIONS:Established method is simple,rapid,accurate and can be used for quality control of C. monnieri liniment.

Simultaneous Determination of Chlorogenic Acid,Baicalin and Imperatorin in Biyanling Tablets by HPLC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of chlorogenic acid, baicalin and imperatorin in Biyanling tablets. METHODS:HPLC method was adopted. The separation was performed on a Waters SunFire C18 column with mobile phase consisted of methanol-0.20% phosphoric acid(gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm and column temperature was 30 ℃. The sample size was 20 μ L. RESULTS:Linear ranges of chlorogenic acid,baicalin and imperatorin were 1.618-16.18 μg/mL,1.624-16.24 μg/mL and 1.762-17.62 μg/mL(r were 0.999 9),respectively. The limits of quantitation were 2.67,2.03,1.87 μg/mL,and the limits of detection were 1.34,1.06, 0.98 μ g/mL,respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 98.01%-101.80%(RSD＝1.38%,n＝9),98.38%-101.40%(RSD＝1.26%,n＝9)and 98.06%-101.40%(RSD＝1.28%,n＝9), respectively. RSDs of tolerance tests were all lower than 2.0%. CONCLUSIONS:The method is simple,accurate,precise,stable, reproducible and durable. It can be used for simultaneous determination of hlorogenic acid,baicalin and imperatorin in Biyanling tablets.

Simultaneous Determination of Isoalantolactone,Alantolactone,Xanthotoxol,Oxypeucedanin, Imperator-in and Osthole in Kangfu Ointment by HPLC / 中国药师

Objective:To establish an HPLC gradient elution method for the determination of isoalantolactone,alantolactone,xan-thotoxol,oxypeucedanin,imperatorin and osthole in Kangfu ointment simultaneously. Methods:Isoalantolactone,alantolactone,xan-thotoxol,oxypeucedanin,imperatorin and osthole were determined by HPLC with a chromatographic column of Agilent TC-C18(250 mm ×4.6 mm,5 μm),the mobile phase was methanol-0.1% formic acid solution with gradient elution at a flow rate of 0.9 ml·min-1. The detection wavelengths were 220 nm and 300 nm. The column temperature was 35℃. Results: The linear range of isoalantolac-tone,alantolactone,xanthotoxol,oxypeucedanin,imperatorin and osthole was 6.16-123.20 μg·ml-1,3.78-75.60 μg·ml-1,1.87-37.40 μg·ml-1,4.06-81.20 μg·ml-1,9.27-185.40 μg·ml-1and 13.89-277.80 μg·ml-1,respectively. The correlation coef-ficient was 0.999 4,0.999 8,0.999 6,0.999 5,0.999 9 and 0.999 7, respectively. The average recovery was 98.04% (RSD=1.06%),97.10%(RSD = 1.53%), 96.73% (RSD = 0.90%), 98.92% (RSD = 1.36%), 99.12% (RSD = 0.83%) and 100.27%(RSD=0.58%),respectively. Conclusion:The method is simple and accurate,which can be used to improve the quality standard for Kangfu ointment.

Vasodilatation Effects and Mechanism of Imperatorin Derivative OW1 / 中国药学杂志

OBJECTIVE To investigate vasodilation effect and mechanism of OW1 was investigated on rat aorta, renal artery, cerebral middle artery, coronary artery, pulmonary artery and mesenteric artery. METHODS Isometric tension was measured in a 610 M-DMT Wire Myograph System. 60 mmol·L-1 KCl was added to the baths to induce pre-contraction of the 6 arteries. The concentration-response curves of OW1 were constructed on endothelium-denuded aorta rings or blocking endothelial, beta adrenergic receptor, three potassium channels and changing the concentration of intracellular or external calcium respectively. RESULTS OW1 could relax the 6 arteries completely, and Emax were all above 95%. pEC50 were (6.11±0.09), (5.76±0.01), (6.22±0.08), (5.78±0.05), (5.65±0.01), and (6.59±0.07) respectively. β-Adrenoceptor, ATP sensitive potassium channel and inwardly rectifying potassium channel were not involved in the vasodilatation, whereas blockage of calcium-activated potassium channel with tetraethylammonium had effect. OW1 could inhibit the influx of extracellular calcium and the release of intracellular calcium, thereby inhibiting vasoconstriction. CONCLUSION OW1 could relax the six arteries. The possible mechanisms of the vasodilatation are mainly involved with inhibiting voltage dependent calcium channel and receptor-mediated Ca2+ influx and release, and might be partly due to opening calcium-activated potassium channel.