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1.
Article in Chinese | WPRIM | ID: wpr-940187

ABSTRACT

ObjectiveTo identify the chemical constituents of Alismatis Rhizoma before and after processing with salt-water by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and to investigate the changes of terpenoids in Alismatis Rhizoma before and after processing with salt-water. MethodUPLC-Q-TOF-MS was used to detect with 0.1% formic acid aqueous solution (A)-acetonitrile (B)as mobile phase for gradient elution (0-0.01 min, 20%B; 0.01-5 min, 20%-40%B; 5-40 min, 40%-95%B; 40-42 min, 95%B; 42-42.1 min, 95%-20%B; 42.1-45 min, 20%B), electrospray ionization (ESI) was selected for collection and detection in positive ion mode with the scanning range of m/z 100-1 250 and ion source temperature at 500 ℃. The data were analyzed by PeakView 1.2.0.3, the components were identified according to the primary and secondary MS data, and combined with the reference substance and literature. After normalized treatment by MarkerView 1.2.1, the MS data were analyzed by principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), and then the differential components before and after processing were screened. The content changes of differential components were analyzed according to the relative peak area. ResultA total of 30 components were identified under positive ion mode, including 28 prototerpene triterpenes and 2 sesquiterpenes. The results of PCA and OPLS-DA showed that there were significant differences in components from Alismatis Rhizoma before and after processing with salt-water, and 10 differential components (alisol B 23-acetate, alisol I, alismol, 11-deoxy-alisol B 23-acetate, alisol B, alisol C, 11-deoxy-alisol B, alisol G, 11-deoxy-alisol C and alisol A) were screened, and the contents of alisol G and alisol A decreased significantly after processing. ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents in raw and salt-processed products of Alismatis Rhizoma. It takes a great difference in the contents of chemical constituents before and after processing, and the difference of substituents is the main reason for this differences, which can provide reference for determining the material basis of efficacy changes of Alismatis Rhizoma before and after processing with salt-water.

2.
Article in Chinese | WPRIM | ID: wpr-940183

ABSTRACT

ObjectiveTo evaluate the success of Qi-deficiency model of Balb/C-nu mice established by swimming exhaustion test from the view of biomarkers and metabolic pathways by metabonomics. MethodBalb/C-nu mice were randomly divided into the normal group and Qi-deficiency group, Qi-deficiency model was established by swimming with 5% body weight metal fixed at the tail for 15 consecutive days until exhaustion (nose tip immersion time>5 s). The urine metabonomics was analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and the mobile phase was acetonitrile (A)-0.1% formic acid aqueous solution (B) for gradient elution (0-1 min, 5%-8%A; 1-4 min, 8%-8.5%A; 4-5 min, 8.5%-12%A; 5-10 min, 12%-40%A; 10-12 min, 40%-100%A; 12-15 min, 100%A), the flow rate was 0.3 mL·min-1, the injection volume was 10 μL, electrospray ionization (ESI) in positive and negative ion modes was used in MS analysis, the MS data were acquired in full-scan mode from m/z 50-1 000. Principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), human metabolome database (HMDB), high collision energy ion fragments collected by MSE and tandem MS (MS/MS) ion information were used to identify potential biomarkers. Kyoto Encyclopedia of Genes and Genomes (KEGG) database and MetaboAnalyst 5.0 were used to analyze the corresponding metabolic pathways and pathway enrichment of biomarkers. ResultEndogenous substances in urine of mice in normal group and Qi-deficiency group were obviously separated, and there were 24 biomarkers with significant difference. The metabolic pathways involved in these biomarkers were tricarboxylic acid cycle, glycolysis metabolism, amino acid metabolism, fatty acid metabolism, pyrimidine metabolism, steroid hormone biosynthesis and tryptophan metabolism. Among them, the metabolic pathways related to energy were tricarboxylic acid cycle, glycolysis metabolism, amino acid metabolism, fatty acid metabolism, pyrimidine metabolism and steroid hormone biosynthesis. ConclusionThrough the investigation of urine metabonomics, combined with the physical signs, the Qi-deficiency model established by swimming exhaustion test in Balb/C-nu mice is successfully evaluated, and it is also verified that there is a close correlation between Qi-deficiency and energy metabolism.

3.
Article in Chinese | WPRIM | ID: wpr-940155

ABSTRACT

ObjectiveTo identify the chemical constituents of Alismatis Rhizoma before and after processing with salt-water by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and to investigate the changes of terpenoids in Alismatis Rhizoma before and after processing with salt-water. MethodUPLC-Q-TOF-MS was used to detect with 0.1% formic acid aqueous solution (A)-acetonitrile (B)as mobile phase for gradient elution (0-0.01 min, 20%B; 0.01-5 min, 20%-40%B; 5-40 min, 40%-95%B; 40-42 min, 95%B; 42-42.1 min, 95%-20%B; 42.1-45 min, 20%B), electrospray ionization (ESI) was selected for collection and detection in positive ion mode with the scanning range of m/z 100-1 250 and ion source temperature at 500 ℃. The data were analyzed by PeakView 1.2.0.3, the components were identified according to the primary and secondary MS data, and combined with the reference substance and literature. After normalized treatment by MarkerView 1.2.1, the MS data were analyzed by principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), and then the differential components before and after processing were screened. The content changes of differential components were analyzed according to the relative peak area. ResultA total of 30 components were identified under positive ion mode, including 28 prototerpene triterpenes and 2 sesquiterpenes. The results of PCA and OPLS-DA showed that there were significant differences in components from Alismatis Rhizoma before and after processing with salt-water, and 10 differential components (alisol B 23-acetate, alisol I, alismol, 11-deoxy-alisol B 23-acetate, alisol B, alisol C, 11-deoxy-alisol B, alisol G, 11-deoxy-alisol C and alisol A) were screened, and the contents of alisol G and alisol A decreased significantly after processing. ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents in raw and salt-processed products of Alismatis Rhizoma. It takes a great difference in the contents of chemical constituents before and after processing, and the difference of substituents is the main reason for this differences, which can provide reference for determining the material basis of efficacy changes of Alismatis Rhizoma before and after processing with salt-water.

4.
Article in Chinese | WPRIM | ID: wpr-940151

ABSTRACT

ObjectiveTo evaluate the success of Qi-deficiency model of Balb/C-nu mice established by swimming exhaustion test from the view of biomarkers and metabolic pathways by metabonomics. MethodBalb/C-nu mice were randomly divided into the normal group and Qi-deficiency group, Qi-deficiency model was established by swimming with 5% body weight metal fixed at the tail for 15 consecutive days until exhaustion (nose tip immersion time>5 s). The urine metabonomics was analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and the mobile phase was acetonitrile (A)-0.1% formic acid aqueous solution (B) for gradient elution (0-1 min, 5%-8%A; 1-4 min, 8%-8.5%A; 4-5 min, 8.5%-12%A; 5-10 min, 12%-40%A; 10-12 min, 40%-100%A; 12-15 min, 100%A), the flow rate was 0.3 mL·min-1, the injection volume was 10 μL, electrospray ionization (ESI) in positive and negative ion modes was used in MS analysis, the MS data were acquired in full-scan mode from m/z 50-1 000. Principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), human metabolome database (HMDB), high collision energy ion fragments collected by MSE and tandem MS (MS/MS) ion information were used to identify potential biomarkers. Kyoto Encyclopedia of Genes and Genomes (KEGG) database and MetaboAnalyst 5.0 were used to analyze the corresponding metabolic pathways and pathway enrichment of biomarkers. ResultEndogenous substances in urine of mice in normal group and Qi-deficiency group were obviously separated, and there were 24 biomarkers with significant difference. The metabolic pathways involved in these biomarkers were tricarboxylic acid cycle, glycolysis metabolism, amino acid metabolism, fatty acid metabolism, pyrimidine metabolism, steroid hormone biosynthesis and tryptophan metabolism. Among them, the metabolic pathways related to energy were tricarboxylic acid cycle, glycolysis metabolism, amino acid metabolism, fatty acid metabolism, pyrimidine metabolism and steroid hormone biosynthesis. ConclusionThrough the investigation of urine metabonomics, combined with the physical signs, the Qi-deficiency model established by swimming exhaustion test in Balb/C-nu mice is successfully evaluated, and it is also verified that there is a close correlation between Qi-deficiency and energy metabolism.

5.
Article in Chinese | WPRIM | ID: wpr-928019

ABSTRACT

Ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF/MS) was used to investigate the effect of Pterocephalus hookeri on serum metabolism of adjuvant arthritis(AA) model rats induced by complete Freund's adjuvant. After the AA model was properly induced, the serum of rats was collected 30 days after treatment. UPLC-Q-TOF-MS chromatograms were collected and analyzed by principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). The results revealed that compared with the control group, the model group showed increased content of 12 biomarkers in the serum(P<0.05) and reduced content of the other nine biomarkers(P<0.05). P. hookeri extract could recover the above-mentioned 19 biomarkers to a certain range. Pathway enrichment showed that these markers mainly involved eight metabolic pathways, including valine, leucine, and isoleucine degradation, arachidonic acid metabolism, arginine and proline metabolism, glycerol phospholipid metabolism, primary bile acid biosynthesis, bile acid biosynthesis, tryptophan metabolism, and unsaturated fatty acid biosynthesis. The findings of this study demonstrate that P. hookeri extract can regulate metabolic disorders and promote the regression of metabolic phenotype to the normal level to exert the therapeutic effect on AA rats. This study is expected to provide a certain scientific basis for the biological research on the treatment of rheumatoid arthritis by P. hookeri.


Subject(s)
Animals , Arthritis, Rheumatoid/drug therapy , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/pharmacology , Medicine, Tibetan Traditional , Metabolomics , Rats
6.
Article in Chinese | WPRIM | ID: wpr-906524

ABSTRACT

Objective:To analyze the chemical constituents in Euodiae Fructus by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). Method:The chromatographic separation was performed on an ACQUITY UPLC BEH C<sub>18</sub> column (2.1 mm×100 mm, 1.7 μm) with acetonitrile (A)-0.1% formic acid aqueous solution (B) as mobile phase (0-3 min, 6%A; 3-4 min, 6%-10%A; 4-7 min, 10%-12%A; 7-8 min, 12%-14%A; 8-13 min, 14%-15%A; 13-15 min, 15%-20%A; 15-18 min, 20%-30%A; 18-21 min, 30%-49%A; 21-25 min, 49%-51%A; 25-27 min, 51%-73%A; 27-30 min, 73%-80%A; 30-31 min, 80%-100%A; 31-32 min, 100%A) for gradient elution. The column temperature was 35 ℃, and the flow rate was 0.4 mL·min<sup>-1</sup>. Mass spectrometry was performed using an electrospray ionization and data were collected in positive and negative ion modes, and the detection range was <italic>m</italic>/<italic>z</italic> 100-1 200. The chemical constituents in Euodiae Fructus were identified rapidly and comprehensively based on the accurate relative molecular mass and combined with literature data and reference substances. Result:A total of 92 chemical constituents were speculatively identified from the 70% methanol extract of Euodiae Fructus, including 39 alkaloids, 19 flavonoids, 12 limonoids, 20 phenolic acids and 2 organic acids. Among them, 26 compounds were confirmed by the reference substances. Conclusion:The compound types of Euodiae Fructus are multiple and quite different in polarity. The chemical compositions of Euodiae Fructus from different regions and species are similar. The established method is rapid and accurate, with which the chemical compositions of Euodiae Fructus have been identified comprehensively. Therefore, this study provides an experimental reference for further clarifying active and toxic constituents of Euodiae Fructus.

7.
Article in Chinese | WPRIM | ID: wpr-906522

ABSTRACT

Objective:To investigate the intervention of <italic>Hedyotis diffusa</italic> (HDW) on colitis associated cancer (CAC) model mice and explore its mechanism. Method:The CAC mouse model was established by synergistic action of azoxymethane (AOM) and dextran sulfate sodium (DSS). The intervention of HDW on CAC mice was evaluated by disease activity index (DAI), colonic tissue morphology, pathological injury score and tumorigenesis rate. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and multivariate statistical analysis were used to analyze the metabonomics of mice serum and to explore the mechanism of HDW intervention on CAC. Result:HDW could significantly improve the general condition of CAC mice, decrease DAI, colon gross morphological score, histopathological score and tumorigenesis rate. Compared with the normal group, 38 kinds of differential metabolites were screened in the model group, including 11 potential biomarkers, involving 11 main metabolic pathways. HDW could significantly regulate 9 kinds of differential metabolites [niacinamide, uridine, 4-pyridoxic acid, LysoPC (18∶0), LysoPE (0∶0/20∶0), myo-inositol, purine, sphinganine 1-phosphate and tetradecanedioic acid] in the model group, including 2 kinds of potential biomarkers (myo-inositol and niacinamide), and HDW could regulate nicotinate and nicotinamide metabolism and inositol phosphate metabolism. Conclusion:HDW has a therapeutic effect on CAC, which may be achieved by regulation of energy metabolism and glucose metabolism.

8.
Article in Chinese | WPRIM | ID: wpr-905840

ABSTRACT

Objective:To quickly analyze and identify the components in raw and wine-processed products of <italic>Polygonatum cyrtonema</italic> (PC) dried rhizomes by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and then find out the differential components before and after processing. Method:The ACQUITY UPLC BEH C<sub>18</sub> column (2.1 mm×100 mm, 1.8 μm) was used with 0.1% formic acid aqueous solution-acetonitrile as the mobile phase for gradient elution at a flow rate of 0.25 mL·min<sup>-1</sup>. Electrospray ionization was selected for collection and detection in positive and negative ion modes, and the data were analyzed by PeakView 1.2.0.3. According to the retention time, accurate relative molecular weight and fragmentation ion information provided by MS, and combined with the reference substance and literature, the components were identified. After normalized treatment, the MS data of each sample were analyzed with principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), and then the differential components before and after processing were screened according to the principle that variable importance in the projection (VIP) value was >1. Result:A total of 38 components were identified from raw and wine-processed products of PC dried rhizomes, including 15 steroidal saponins, 6 alkaloids, 3 flavonoids, 2 amino acids, 2 organic acids and 10 others. The results of PCA and OPLS-DA showed that there were significant differences in the contents of components in PC dried rhizomes before and after processing, and 16 differential components such as kingianoside Z, disporopsin and linoleic acid were screened. Conclusion:UPLC-Q-TOF-MS technique can accurately and comprehensively identify the components in PC dried rhizomes, these components are mainly steroidal saponins, flavonoids and alkaloids. It takes a great difference in the contents of components before and after processing, and transformation of the same category components is the main reason for the differences of raw and wine-processed products, which will provide reference for the researches on material basis and processing chemistry of PC dried rhizomes.

9.
Article in Chinese | WPRIM | ID: wpr-873066

ABSTRACT

Objective:To compare the effects of different processing techniques on the chemical constituents of Aurantii Fructus for screening the dominant decoction pieces. Method:UPLC-Q/TOF-MS was used to detect the chemical constituents of Aurantii Fructus, chromatography separation was achieved on an ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm), and gradient elution was performed with 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) as mobile phase (0-10 min, 5%-35%B; 10-18 min, 35%-75%B; 18-21 min, 75%-100%B; 21-24 min, 100%B; 24-24.1 min, 100%-5%B; 24.1-28 min, 5%B). Data acquisition was carried out in electrospray ionization (ESI) under the positive ion mode, the scanning range was m/z 50-1 200. The chemical constituents in methanol extract of Aurantii Fructus were identified according to reference substance, relative molecular weight, mass spectrometric cleavage rule and literature information. SIMCA-P 13.0 software was used to establish principal component analysis (PCA) model and partial least squares-discriminant analysis (PLS-DA) model of Aurantii Fructus processed products, PCA score plot, PLS-DA loading plot and variable importance in the protection (VIP) values were obtained to screen the material basis for the main differences before and after processing of Aurantii Fructus. Result:A total of 54 chemical components were identified by UPLC-Q/TOF-MS. PCA indicated that there were significant differences among different groups of Aurantii Fructus processed by different methods. A total of 14 chemical components with VIP value >1 were screened by PLS-DA as the main chemical markers for the differences before and after processing, including hesperidin, poncirin, narirutin, etc. The comprehensive weighted score showed that the content of effective components in Aurantii Fructus processed with honey bran was the highest. Conclusion:The contents of chemical constituents in Aurantii Fructus before and after processing are significantly changed. Flavonoids are the most important compounds to distinguish different processed products of Aurantii Fructus. Aurantii Fructus processed with honey bran is the dominant variety.

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