Study on Metabolism of Miao Medicine Laportea bulbifera Extract in Isolated Human Intestinal Flora / 中国药房

OBJECTIVE：To explore the metabolic charact eristics of Miao medicine Laportea bulbifera extract in isolated human intestinal flora. METHODS ：L. bulbifera was extracted with 70％ ethanol reflux extraction. After concentration，extraction with n-butanol and drying ，L. bulbifera extract was obtained. Taking 0.05 g/mL L. bulbifera extract 1 mL mixed with isolated human intestinal flora fluid 10 mL and cultured for 36 h in anaerobic environment （setting up blank control without drugs or human intestinal bacterial solution ），so as to simulate the metabolic process of the extract in human intestine. The metabolites were detected by UPLC-Q-TOF/MS. The determination was performed on Agilent Eclipse Plus C 18 RRHD column with mobile phase consisted of 0.01％ formic acid water solution- 0.01％ formic acid acetonitrile solution （gradient eluetion ）at the flow rate of 0.25 mL/min. The column temperature was set at 40 ℃，and the sample size was 1 µL. ESI detection was adopted and scanned by negative ion mode （ESI－）；the capillary voltage was 4.5 kV，the ion source temperature was 120 ℃，the collision energy was 15-32 V，and the scanning range was m/z 50-1 000. The “Strip”module of MassLynx V 4.1 software was used to analyze the differential chromatograms between the reaction solution and the blank control of L. bulbifera extract. Mass spectrum data and UNIFI so ftware were used to predict relative molecular weight and formula ；based on the information of substance control and related literature reports ， the structure and biotransformation pathway of L. bulbifera metabolites in isolated human intestinal flora were predicted and analyzed. RESULTS & CONCLUSIONS ： A total of 3 prototype ： products（rutin，quercetin，kaempferol-3-O-rutinoside）and 22metabolites （mainly the metabolites of quercetin ，mono- caffeoylquinic acid ，isoquercitrin，etc.） were detected after metabolized in isolated human intestinal flora. Itsbiotransformation pathway is phase Ⅰ reaction，which mainly consisted of reduction ，oxidation and hydrolysis.

Pharmacokinetics of three index components in flower of Polygonumorientale L.in rat plasma by UPLC-MS / 中国药理学通报

Aims To establish a UPLC-MS method for quantifying protocatechuie acid,kaempferol-3-O-β-D-glucoside and quercitrin and to investigate the pharma-cokinetics of three indix components in flower of Poly-gonumorientale L.in rat plasma.Methods The anal-ysis was achieved by BEH C18 column (2.1 mm ×100 mm,1.7 μm)with a mobile phase composed of 0.1%formic acid using step gradient elution.A TQD tandem mass spectrometry equipped with electrospray ionization source was used as detector and operated by selected ion recording(SIR)mode.Results In the selected linear range,calibration curves of the three markers components showed good linearity.Extraction recovery rate,precision,accuracy and stability reached the de-termination request.The parameters of Tmax (h ) in three index components were 0.46 ±0.1,0.79 ±0.33 and 2.63 ±4.6,respectively;Cmax (μg·L -1 )in three index components were 463.8 ±207.81,18.53 ±7.82 and 137.38 ±71.09,respectively.Conclusion The fully validated UPLC-MS method has been successfully applied to the pharmacokinetic study of the three index components in flower of Polygonumorientale L.in rat plasma.

Simultaneous determination of human plasma protein binding of bioactive flavonoids in Polygonum orientale by equilibrium dialysis combined with UPLC-MS/MS / 药物分析学报

A simple and selective ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) assay was developed for the determination of the human plasma protein binding of four bioactive flavonoids (such as orientin and vitexin) in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C18 column with a mobile phase composed of 0.1%formic acid in acetonitrile and 0.1%aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITY?TQD system was operated under the multiple reaction monitoring (MRM) mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations (r40.99) of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

Simultaneous determination of isoorientin, scutellarin and cynaroside in rat plasma and pharmacokinetics by UPLC-MS/MS / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a UPLC-MS/MS analysical method for simultaneous determination of concentrations of isoorientin, scutellarin and cynaroside in rat plasma and to study their pharmacokinetic characteristics after intravenous injection of 3 doses of Fufang Hongcao in rats.</p><p><b>METHOD</b>Acidified plasma samples were precipitated for protein with methanol. Waters Acquity BEH C18 column was adopted for spectrum, with mobile phase as 0. 1% formic acid acetonitrile-0. 1% formic acid-water gradient elution. Detection was carried out by the multiple reaction monitoring (MRM) positive ion mode with ESI ionization source.</p><p><b>RESULT</b>Three flavonoids show a good linear relationship, with the extraction recovery ranging between 78.56% and 101.91% and a high intra-and inter-day precisions and accuracy. The MRT of the three flavonoids were all lower than 22 min in rats.</p><p><b>CONCLUSION</b>The above men tioned method is so specific, rapid, sensitive that it is suitable for pharmacokinetic studies of Fufang Hongcao injection in rats.</p>

Chemical constituents in herb of Polygonum orientale II / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of the essential substance from the Polygonum oriental.</p><p><b>METHOD</b>Chromatographic techniques were employed for isolation and purification of the constituents and their structures were determined by spectral analysis and chemical evidence.</p><p><b>RESULT</b>Seven compounds were obtained and identified as ombuine-3-O-beta-D-galactopyranoside (1), ombuine-3-O-rutinoside (2), tryptophan (3), quercetin-3-O-methyl ether (4), kaempferol-3-O-(2"-O-alpha-L-rhamnopyranosyl) -beta-D-glucuronopyranoside (5), quercetin-3-O-(2"-O-alpha-L-rhamnopyranosyl)-beta-D-glucuronopyranoside (6), quercetin-3-O-beta-D-glucuronide (7).</p><p><b>CONCLUSION</b>Compounds 4-7 were isolated from P. oriental for the first time and compounds 1-3 were firstly obtained from the genus Polygonum. The total 1H and 13C-NMR date of compound 1 were assigned for first time.</p>

Design, synthesis and anti-oxidative evaluation of L-amino acid prodrugs of scutellarein / 药学学报

To design and synthesize a series of novel scutellarein 4'-L-amino acid prodrugs with more potent anti-oxidative activity and improved physicochemical properties. Scutellarein was used as lead compound, according to successful experience of improving bioavailability of oral administration drugs by active transport mechanism, principle of hybridization was used to introducing L-amino acid structural fragments at 4'-position of scutellarein to design and synthesize target scutellarein 4'-L-amino acid prodrugs. The synthetic compounds were tested on their physicochemical properties and in vitro anti-oxidative activity against H202 induced oxidative damage in PC12 cells. Five compounds were found to have more potent anti-oxidative activity than positive control VE. Moreover the physicochemical properties of synthesized compounds were evaluated, and the results revealed that L-amino acid ether derivatives are more stable (t1/2 9-92 h) than their corresponding ester derivatives (t1/2 0.5 h). Water solubility of scutellarein 4'-L-amino acid ester and ether derivatives were 1 796-4 100 microg.mL-1 and 27.7-81.1 microg.mL-1 respectively, in comparison with scutellarin, the solubility of compounds 18, 19 and 22, 24-27 increased about 120-280 fold and 2-6 fold respectively. All these results suggested that L-amino acid prodrug strategy has significant potential in scutellarein prodrug design.

Design, synthesis and anti-hBV evaluation of adefovir mono-L-amino acid ester, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs / 药学学报

A series of adefovir mono-L-amino acid esters, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs with more potent anti-HBV activity and lower nephrotoxicity were designed and synthesized. Adefovir bis (L-amino acid) ester was used as lead compound, according to pathological and pharmacological findings that non-steroidal anti-inflammatory drugs can effectively inhibit the organic anion transporter 1 (hOAT1)-mediated adefovir phosphonic acid pairs of anion transport across tubular basement membrane thereby reducing the nephrotoxicity of adefovir. Flatten design principle was used to introducing non-steroidal anti-inflammatory drugs structural fragments to design and synthesize target adefovir mixture ester prodrugs. HepG2 2.2.15 cell line was used as in vitro anti-HBV activity evaluation model. Five compounds exhibited antiviral activity, and compound 18 showed the most potent anti-HBV activity and relatively high selective index (EC50 3.92 micromol L(-1), SI 9.97). HK-2 cell line was used as in vitro model to evaluate nephrotoxicity. Results suggested the target compounds have lower cytotoxicity than the positive control. Moreover, by analyzing the primary structure and activity relationship of these compounds, it could suggest that mono-L-amino acid ester, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs strategy has significant potential in the acyclic nucleoside phosphonates prodrug design.

Chemical constituents of flowers from Polygonum orientale / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in the active portion from the flowers of Polygonum oriental.</p><p><b>METHOD</b>Chromatographic techniques were employed for isolation and purification of the constituents and their structures were determined by spectral analysis and chemical evidence.</p><p><b>RESULT</b>Nine compounds were obtained and identified as alphitonin (1), methyl 3, 4-dihydroxybenzoate (2), apocynin (3), kaempferol-3-O-beta-D-glucoside (4), 1,3,5-trihydroxybenzene (5), 3,3'-dimethoxyellagic-acid-4-O-beta-D-glucoside (6), kaempferol-3-O-alpha-L-rhamnoside (7), quercetin-3-O-alpha-L-rhamnoside (8), kaempferol (9).</p><p><b>CONCLUSION</b>Compounds 2, 4, 5 were isolated from P. oriental for the first time and compounds 1, 3 were firstly obtained from the genus Polygonum.</p>

Studies on extraction of Ligusticum chuanxiong in Hongye Xintong soft capsule by super critical fluid extraction / 中国中药杂志

<p><b>OBJECTIVE</b>To study the rationality of extraction prosess for the Ligusticum chuanxiong in Hongye Xintong Soft Capsule by super critical fluid extraction(SFE).</p><p><b>METHOD</b>Conditions for the extraction were optimized by orthogonal experimental design as guided by the extraction rate and content of ligustilid in the extract; Combined with the experiment of rats ligated the left coronary artery, the two compounds containing different extracts (steam distillation and SFE) were compared to determine the extraction prosess.</p><p><b>RESULT</b>The best extraction conditions were established as following: pressure 30 MPa, temperature 50 degrees C, extracting time 4 h, separate pressure 10 MPa. separate temperature 40 degrees C. Compared with steam distillation, the compound containing SFE extract showed stronger protective effects on rats ligated the left coronary artery.</p><p><b>CONCLUSION</b>Super critical fluid can extract active ingredients in Ligusticum chuanxiong effectively.</p>

Determination of baicalin in Fushuye by HPLC / 中国药学杂志

OBJECTIVE:To domminate the quality of Fushuye,the contents of baicalin in Fushuye was determined by HPLC.METHOD:Baicalin was separated on Spherisorb C18 column and detected at 278 nm,by using methanol-4g.L-1 phosphoric acid (50∶50) as mobile phase.RESULTS:The resolution was 4.5.The number of theoretical plates calculaled for baicalin peak was 3800.The standard curve was linear over the range of 20 ng～900ng,with the correlation coefficient 0.999 9.The average recovery was 99.88% with RSD=1.54%(n=5). CONCLUSION:The method is simple and fast,and can be used for quantitative analysis of Fushuye.

Pattern recognition of the fingerprint of Polygonum orientale in Guizhou Province / 中成药

AIM: To study pattern recognition of the fingerprint of Polygonum orientale from Guizhou Province. METHODS: 20 batches of samples,Polygonum orientale,come from different producing areas and harvest time were carried out to gain fingerprints by HPLC-DAD,as compared with retention time and ultraviolet spectra of standard substances.The pattern recognition of the characteristic fingerprint of Polygonum orientale,known as cluster analysis and principal component analysis,formed. RESULTS: The fingerprint of 20 batches of Polygonum orientale showed 12 characteristic peaks,in which 9 common peaks were confirmed.the samples were grouped into 3 types of harvest time. CONCLUSION: The quality of Polygonum orientale in Guizhou Province is stable,the pattern recognition of the fingerprint provides the experimental basis for manufacture and quality control of Polygonum orientale.

HPLC fingerprint analysis of Herba Polygoni Orientalis / 中成药

AIM: To study and establish the fingerprint of raw Herba Polygoni Orientalis by RP-HPLC. METHODS: The chromatographic conditions were as follow: an Inersil-ODS-3 column was used;the mobile phase was composed of acetontrile and 0.1% phosphoric acid with gradient elution;the flow was 1.0 mL?mim~(-1) and the UV absorbance detection was set at 300 nm. RESULTS: Under the selected chromatographic conditions. Similarity of 10 batches of good HPLC fingerprint of raw Herba Polygoni Orientalis were obtained. no less than 0.9.( CONCLUSION): Quality of raw Herba Polygoni Orientalis can be controlled effectively by HPLC-UV fingerprint.