Study on effective sites of anti-gonococcus in Polygonum capitatum extract / 中草药

Objective To analyze the components of active anti-gonococcal fractions in the extracts of Polygonum capitatum. Methods Drug sensitive paper method and Neisseria gonorrhoeae culture method were used to detect the antibacterial activity of different parts of the flower bud extract against Neisseria gonorrhoeae. UPLC-TOF-MS was used to identify the effective parts of the flower bud extract with anti-gonococcal effect. Results As a result, in the eluate obtained by extracting with a specific macroporous resin, the hydrolysable tannin was contained in the methanol (35%) eluate (fraction F3), such as trigallocyl glucose. This fraction had a good anti-gonococcal effect. The fractions of F2 and F4-6 had a weak antibacterial effect, while other fractions have no antibacterial effect. Conclusion This study provides a basis for the use of trigalactosyl glucose in the preparation of anti-gonococcal drugs, and lays a foundation for the development of a clinical drug of P. capitatum extracts against Neisseria gonorrhoeae.

Untargeted Urinary Metabolomic Study on Toxicity-alleviation Effect of Huangqi Oral Solution in Cisplatin-exposed Rats / 分析化学

A liquid chromatography-quadrupole-time-of-flight mass spectrometer(LC-Q/TOF-MS) based urinary metabolomic approach was employed to assess the toxicity-alleviation effect of Huangqi oral solution(HOs) on cisplatin-exposed rats and explore its possible mechanisms. Rat toxicity model was developed by multiple intraperitoneal injection of low-dose cisplatin, while HOs was orally administrated to rats simultaneously for 16 consecutive days to attenuate or reduce the cisplatin-induced toxicity. 24-hour urine samples on day 18 were collected and analyzed using LC-Q/TOF-MS to obtain the dataset of urinary metabolites. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to assess the quality of the dataset and screen the potential toxicity-alleviation biomarkers. The serum level of rat creatinine and urea nitrogen on day 20 was determined, and the results showed that successive administration of HOs significantly reduced the cisplatin-induced increase of creatinine and urea nitrogen. PCA cluster analysis clearly demonstrated that HOs could partly improve the CDDP-induced abnormality of metabolic profiling. 35 urinary metabolites were finally screened as the potential biomarkers associated with the toxicity-attenuation effect of HOs, according to the combination of the analysis results of OPLS-DA, t-test and fold change analysis. Further metabolic pathway analysis revealed that HOs could restore the metabolic disorders of amino acid, energy and nucleotide, thereby exerted its toxicity-alleviation effect.

Plasma Metabonomics Research of Astragalus Flavonoids Intervention on Rats of Dampness Stagnancy Due to Spleen Deficiency / 分析化学

A method of high performance liquid chromatography coupled with time-of-flight mass spectrometry was used to detect the endogenous metabolites changes in plasma of normal rats, rats of dampness stagnancy due to spleen deficiency and Astragalus flavone component intervention rats. Metabolism map of rat plasma was obtained and the mechanism of Astragalus flavonoids on dampness stagnancy due to spleen deficiency was studied. Rat model with dampness stagnancy due to spleen deficiency was established by high fat and low protein diet plus load swimming. Liquid chromatography tandem mass spectrometry was used for the analysis of rat plasma sample, and 0.05％ formic acid water with 0.05％ formic acid acetonitrile as the mobile phase was applied in gradient elution with Halo C18 chromatographic column. In this study, partial least squares discriminant analysis and variance analysis were used to screen the potential biomarkers, it was found that the metabolic profile of the Astragalus flavonoids was different from that of the model group, which was close to that of the normal group. A total of 11 potential biomarkers were identified, including glycerol phospholipids, sphingolipids, amino acids, and so on. The metabolic pathways of biomarkers including three tricarboxylic acid cycle, glycerol phospholipid metabolism, fatty acid metabolism, amino acid metabolism and so on, which mainly related to energy metabolism and fat metabolism in the body. Related indexes of rats with syndrome of spleen deficiency of water and dampness were significantly callback after Astragalus flavone intervention, including macro indicators such as body weight, independent activities and micro indicators such as metabolic markers, blood lipids and others. The result showed that Astragalus flavonoids played the role of strengthening the spleen and draining the water mainly through regulating the energy metabolism, fat metabolism and so on.

Evaluation of Brodifacoum-induced Toxicity by Metabonomics Approach Based on HPLC-TOF-MS / 法医学杂志

OBJECTIVES@#To analyse the metabolic changes in urine of rats with brodifacoum intoxication, and to reveal the molecular mechanism of brodifacoum-induced toxicity on rats.@*METHODS@#By establishing a brodifacoum poisoning rats model, the urine metabolic profiling data of rats were acquired using high performance liquid chromatography-time of flight mass spectrometry （HPLC-TOF-MS）. The orthogonal partial least squares analysis-discrimination analysis （OPLS-DA） was applied for the multivariate statistics and the discovery of differential metabolites closely related to toxicity of brodifacoum.@*RESULTS@#OPLS-DA score plot showed that the urinary metabolic at different time points before and after drug administration had good similarity within time period and presented clustering phenomenon. Comparing the urine samples of rats before drug administration with which after drug administration, twenty-two metabolites related to brodifacoum-induced toxicity were selected.@*CONCLUSIONS@#The toxic effect of brodifacoum worked by disturbing the metabolic pathways in rats such as tricarboxylic cycle, glycolysis, sphingolipid metabolism and tryptophan metabolism, and the toxicity of brodifacoum is characterized of accumulation effect. The metabonomic method based on urine HPLC-TOF-MS can provide a novel insight into the study on molecular mechanism of brodifacoum-induced toxicity.

HPLC-TOF/MS in identifying steroidal saponins in rat urine after oral administration of Paris polyphylla extract / 第二军医大学学报

Objective To use high-performance liquid chromatography-time of flight mass spectrometry (HPLC-TOF/ MS) for examining the steroidal saponins in rat urine after oral administration of Paris polyphylla extract, so as to lay a foundation for studying the metabolism of steroidal saponins in vivo. Methods SD rats were administered with an oral dose of 1 6 g Paris polyphylla extracts/kg body weight. The urine samples were collected 24 h after administration by oral gavage. The sample analysis was carried out on a reverse phase MG-C18 column (3. 0 mm× 100 3. 0μm) using a gradient mobile phase system of acetonitrile-water containing 0. 1% formic acid. TOF/MS was applied for qualitative analysis under positive and negative ion modes. The steroidal saponins in rat urine were identified by using the accurate molecular weight obtained by TOF/ MS and formula database. Results A total of 20 steroidal saponins were identified in the rat urine, and two pairs of isomers were deduced through their fragment ions and standards: polyphyllin VI and pennogenin-3-0-α-L-rhamnopyranosyl(l →4)-α-L-rhamnopyranosyl (1 → 3)-[-α--rhamnopyranosyl (1 → 2)]-β-D-glucopyranoside; and gracillin and pennogenin-3-O-α-L-rhamnopyranosyKl → 4)-α-L-rhamnopyranosyl (1 → 4)-β-D-glucopyranoside. Conclusion The established analysis method is accurate, reliable for identifying steroidal saponins in vivo, which paves a way for further pharmacokinetics and pharmacodynamics study of Paris polyphylla.

HPLC-TOF/MS in identification of chemical constituents of Fuzhengpingxiao Capsule / 第二军医大学学报

Objective To use high-performance liquid chromatography-time of flight mass spectrometry (HPLC-TOF/ MS) for analyzing the chemical constituents of Fuzhengpingxiao Capsule. Methods The separation was performed on a Agilent Eclipse plus C18 reverse phase column (4. 6 mm×250 mm, 5 μm). The mobile phase consisted of water containing 0. 1% formic acid and acetonitrile was used as gradient elute. The flow rate was 1 ml/min, the post-column split ratio was 3:1, and the temperature of column was 25°C. Time-of-flight mass spectrometer(TOF/MS) and electrospray ion source (ESI) was employed for qualitative analysis under both positive and negative ion mode, and mass scan range was m/z 100-1 100. Results A total of 247 major chemical constituents were identified in Fuzhengpingxiao Capsule by HPLC-TOF/MS, including 168 in positive mode, 103 in negative mode, and 24 in both. The source herb of the components were identified. Conclusion An efficient HPLC-TOF/MS approach has been established for studying the chemical constituents in Fuzhengpingxiao Capsule, which paves a way for the quality control and further in vivo studies of the preparation.

RRLC-TOF/MS in identification of constituents and metabolites of Radix Saposhnikoviae in rat plasma and urine / 第二军医大学学报

Objective: To analyze the constituents and metabolites of Radix Sa poshnikoviae (RS) in rat plasma and urine by rapid-resolution liquid chromatography-time of flight mass spectrometry (RRLC-TOF/MS), so as to explore the active ingredients and metabolites of RS in vivo. Methods: The separation was performed on a Angilent Zorbax Extend-C14 (5 μm, 250 mm x 4.6 mm id) column, with a methanol-water mobile phase system used for gradient elution. Time-of-flight mass spectrometer (TOF/MS) was applied for qualitative analysis under positive ion mode. Based on the accurate molecular weight of TOF/MS detection and the compound list of RS established previously, the constituents and metabolites of RS in different matrix in vivo were identified. Results: Six constituents of RS were identified in the plasma, sucrose, prim-O-glucosylcimifugin, cimifugin, nodakenetin, 5-O-methylvisamminol, and 3′-O-i-butyrylhammaudol. Eight constituents were identified in the urine, prim-O-glucosylcimifugin, divaricatacid, cimifugin, 4′-O-glucosyl-5-O- methylvisamminol, (3S)-2,2-dimethyl-3, 5-dihydroxy-8-hydroxymethyl-3, 4-dihydro-2H, 6H-benzo-[1, 2-b: 5, 4-b′] dipyran-6-one, 5-O-methylvisamminol, see-O-β-D-glucosylhammaudol, and wogonin. Two metabolites were identified in the urine, glucuronide of cimifujin and an isomer of it. Conclusion: The present method is reliable and effective for identifying compounds of RS in vivo, and it can provide a reference and evidence for the further pharmacodynamics experiments.