Anti-depressant components and mechanism of Rehmanniae Radix based on UPLC-Q-Orbitrap HRMS and network pharmacology / 中国中药杂志

This study aimed to explore the anti-depressant components of Rehmanniae Radix and its action mechanism based on network pharmacology combined with molecular docking. The main components of Rehmanniae Radix were identified by ultra-high performance liquid chromatography-quadrupole/Orbitrap high resolution mass spectrometry(UPLC-Q-Orbitrap HRMS), and the related targets were predicted using SwissTargetPrediction. Following the collection of depression-related targets from GeneCards, OMIM and TTD, a protein-protein interaction(PPI) network was constructed using STRING. GO and KEGG pathway enrichment analysis was performed by Metascape. Cytoscape 3.7.2 was used to construct the networks of "components-targets-disease" and "components-targets-pathways", based on which the key targets and their corresponding components were obtained and then preliminarily verified by molecular docking. Rehmanniae Radix contained 85 components including iridoids, ionones, and phenylethanoid glycosides. The results of network analysis showed that the main anti-depressant components of Rehmanniae Radix were catalpol, melittoside, genameside C, gardoside, 6-O-p-coumaroyl ajugol, genipin-1-gentiobioside, jiocarotenoside A1, neo-rehmannioside, rehmannioside C, jionoside C, jionoside D, verbascoside, rehmannioside, cistanoside F, and leucosceptoside A, corresponding to the following 16 core anti-depression targets: AKT1, ALB, IL6, APP, MAPK1, CXCL8, VEGFA, TNF, HSP90 AA1, SIRT1, CNR1, CTNNB1, OPRM1, DRD2, ESR1, and SLC6 A4. As revealed by molecular docking, hydrogen bonding and hydrophobicity might be the main action forms. The key anti-depression targets of Rehmanniae Radix were concentrated in 24 signaling pathways, including neuroactive ligand-receptor interaction, neurodegenerative disease-multiple diseases pathway, phosphatidylinositol 3-kinase/protein kinase B pathway, serotonergic synapse, and Alzheimer's disease.

Analysis and Identification of Flavonoids in Citri Reticulatae Pericarpium with Different Aging Time by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective:To analyze and identify the flavonoids of Citri Reticulatae Pericarpium with different aging time by an ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS）. Method:Compounds were separated on Agilent Extend-C₁₈ column （3.0 mm×100 mm， 1.8 μm）， mobile phase was 0.1% acetic acid aqueous solution （A）-0.1% acetic acid methanol solution （B） for gradient elution （0-25 min， 5%-95%B； 25-30 min， 95%B； 30-30.1 min， 95%-5%B； 30.1-35 min， 5%B）， the flow rate was 0.4 mL·min^-1， and the column temperature was set at 30 ℃. High resolution mass spectrometry was performed with electrospray ionization （ESI）， and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS²）. The multistage ion fragment information combined with mzCloud network database， local high resolution mass spectrometry database of traditional Chinese medicine components （OTCML）， literature information and relevant reference materials were used for accurate qualitative analysis. Result:Totally 43 flavonoids in Citri Reticulatae Pericarpium were identified， including 24 flavones， 5 flavonols， 13 dihydroflavones and 1 chalcone. The flavonoids in samples with different aging time were basically consistent in material types， but the peak area was different. According to the comparison of relative content in the peak area， it was found that the relative contents of 30 flavonoids showed an overall increasing trend with the increase of aging time. Among them， the relative contents of 24 flavonoids （such as hesperidin， diosmin， 6-demethoxytangeretin， nobiletin and tangeretin） increased significantly. There was no significant change in the relative contents of the other 13 flavonoids （such as naringenin and neohesperidin）. Conclusion:An efficient method is established in this paper to identify flavonoids in Citri Reticulatae Pericarpium with different aging time and their relative content changes rapidly and accurately. The findings provide a methodological reference for the study on pharmacodynamic material base and quality control of Citri Reticulatae Pericarpium， and it provides experimental basis that drugs processed long time ago have better effect of Citri Reticulatae Pericarpium.

Identification of Chemical Constituents in Coptis chinensis Inflorescence by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective:An ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） was developed to analyze and identify the chemical constituents in <italic>Coptis chinensis</italic> inflorescence. Method:The chromatographic separation was performed on ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-15 min， 10%-22%B； 15-20 min， 22%B； 20-25 min， 22%-44%B； 25-35 min， 44%-50%B； 35-40 min， 50%-60%B； 40-55 min， 60%-85%B）， the flow rate was 0.15 mL·min^-1， the injection volume was 3 μL and the column temperature was 30 ℃. HRMS was equipped with electrospray ionization （ESI） and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS²）. Compound Discoverer 3.0 software combined with mzCloud， mzVault， ChemSpider databases and HRMS database of components in traditional Chinese medicine were used to analyze and identify the collected data by HRMS， based on accurate relative molecular mass， retention time and characteristic ion fragmentation of the compounds， as well as literature information and relevant reference materials. Result:A total of 51 chemical constituents were identified in <italic>C</italic>.<italic> chinensis</italic> inflorescence， including 16 alkaloids， 14 flavonoids， 7 phenylpropanoids， 7 organic acids and 7 others. Among them， 10 components ［berberine， palmatine， coptidine， rutin， quercetin， isoquercitrin， chlorogenic acid， cryptochlorogenic acid，<italic> D</italic>-（-） quinic acid and <italic>D</italic>-proline］ were unambiguously identified by comparing with reference standards. Conclusion:The established UPLC-Q-Orbitrap HRMS can be used to accurately analyze and identify chemical constituents of <italic>C. chinensis</italic> inflorescence. A total of 41 chemical constituents are reported from <italic>C. chinensis</italic> inflorescence for the first time and 6 alkaloids are found from the <italic>C. chinensis</italic> for the first time. These findings can provide methodological reference and experimental basis for the basic research of quality evaluation and efficacy materials of <italic>C. chinensis</italic> inflorescence， and lay a foundation for its further development and utilization.

Analysis and Identification of Chemical Constituents of Citri Sarcodactylis Fructus by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective::To analyze and identify the chemical constituents of Citri Sarcodactylis Fructus by an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbirap HRMS) method. Method::Compounds were separated on Thermo Scientific Accucore™ C18 column (3 mm×100 mm, 2.6 μm). The mobile phase was 0.1% formic acid solution and 0.1% formic acetonitrile solution. The flow rate was 0.3 mL·min-1, and the column temperature was set at 30 ℃. HRMS was performed using an electrospray ion source (ESI), and scanned in a positive ion mode by means of full scan/data dependent secondary scan (Full MS/dd-MS2). Compound Discoverer 3.0 software that could be linked to mzCloud network database and local high-resolution mass spectrometry database of traditional Chinese medicine components was used to analyze the data, based on accurate molecular mass, retention behaviors and characteristic ion fragmentation of the compounds, as well as literature information and relevant reference materials. Result::Totally 54 chemical constituents in Citri Sarcodactylis Fructus were identified, including 16 flavonoids, 17 coumarins, 3 limonoids, 6 nucleosides and nucleobases, 2 organic acids, 3 aromatic aldehydes, 1 amino acid and 6 other components. Conclusion::The established UPLC-Q-Orbitrap HRMS method can be used to effectively and rapidly identify main chemical constituents of Citri Sarcodactylis Fructus. The findings provide a methodological reference and theoretical foundation for defining the pharmacodynamic material base and optimizing quality control index of Citri Sarcodactylis Fructus, which is of guiding significance for further development and utilization of the resources.

Study on Mechanism of Reducing Excess Fire of Liver and Gallbladder of Bile Processed Coptidis Rhizoma Based on UPLC-Q-Orbitrap HRMS and Network Pharmacology / 中国实验方剂学杂志

Objective:Based on UPLC-Q-Orbitrap HRMS and network pharmacology, the material basis, processing principle and molecular mechanism of bile processed Coptidis Rhizoma (BPRC) for reducing excess fire of liver and gallbladder were elucidated.Method:The chemical ingredients of BPRC were analyzed by UPLC-Q-Orbitrap HRMS. Chromatographic separation was achieved with 0.1% formic acid solution (A)-acetonitrile (B) as the mobile phase in gradient elution (0-20 min, 5%-80%B; 20-30 min, 80%-95%B; 30-30.1 min, 95%-5%B; 30.1-35 min, 95%-5%B). The flow rate was 0.2 mL·min-1, electrospray ionization (ESI) was applied and operated in positive and negative ion modes, the acquisition range was m/z 100-1 500. Based on the clinical manifestations and pathogenic factors of excess fire of liver and gallbladder, the potential effective ingredients, targets and functional characteristics of BPRC were predicted and analyzed by online database. Based on the characteristics of the new active ingredients after processing, the processing principle of BPRC was investigated by network pharmacology.Result:A total of 19 ingredients in BPRC were identified, six of which were newly added cholic acids after processing. It was determined that the alkaloids, including worenine, epiberberine, jatrorrhizine, coptisine, berberrubine, berberine, palmatine and cholic acids, including glycohyodeoxycholic acid, taurohyodeoxycholic acid, glycochenodeoxycholic acid, hyodeoxycholic acid and taurochenodeoxycholic acid, were identified as material basis of BPRC. A total of 66 targets of reducing excess fire of liver and gallbladder of BPRC were screened. There were 16 common targets and multiple same signaling pathways between cholic acids and alkaloids of BPRC, and many lesions of excess fire of liver and gallbladder were target organs of cholic acids. By acting on some targets, including albumin (ALB), Caspase-3 (CASP3), mitogen-activated protein kinase 14 (MAPK14), glucocorticoid receptor (NR3C1) and other targets and some signaling pathways, including interleukin (IL)-17, phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), MAPK and other pathways, BPRC could reduce excess fire of liver and gallbladder.Conclusion:BPRC has the characteristics of multi-component, multi-target and multi-pathway on reducing excess fire of liver and gallbladder. Bile and Coptidis Rhizoma have synergistic effect and bile can enhance the intensity of BPRC in lesions, which confirms the processing theory that the effect of BPRC on excess fire of liver and gallbladder enhance after being processed by bile.

Analysis and Identification of Sesquiterpenes in Alpinia oxzphylla Miq Based on UPLC-Q-Orbitrap HRMS / 中草药

Objective: To establish a rapid and accurate analytical method for the identification of complex system of traditional Chinese medicine, and to systematically clarify the chemical composition of sesquiterpenes in Alpinia oxyphylla. Method: On the basis of optimizing the extraction process of sesquiterpenes, the accurate molecular weight and secondary fragment ions information of unknown compounds were captured by ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Compared with the relative retention time and mass spectrometry data of the reference substance, combined with relevant references and databases, the sesquiterpene unknown compounds in the fruits of A. oxyphylla were accurately and rapidly characterized. Results: A total of 24 sesquiterpenes were identified and classified into four categories according to their skeleton structure, including nine eudesmane-type, six cadinane-type, eight eremophilane-type, and one oplopanone-type. Conclusion: In this study, the established analytical method was used to realize the rapid and accurate identification of sesquiterpenes in the fruits of A. oxyphylla, which provided a theoretical basis for the research on the pharmacodynamic substance basis and quality control of the fruits of A. oxyphylla.

The Chemical Constituents Study of Naoxintong Capsules Based on UHPLC-Q-Orbitrap HRMS / 中国药学杂志

OBJECTIVE: To establish a novel ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) method to study the chemical composition of Naoxintong capsules rapidly and accurately. METHODS: The information of accurate mass in full-MS and multistage fragment ions was obtained by the novel UHPLC-Q-Orbitrap technology. Chemical constituents were identified by comparing the relative retention time and the mass data of the reference substances, meanwhile consulting the reference literature or the Mass Bank, Chemical Book network database. RESULTS: Forty-four compounds were finally identified in this study, mainly including flavones, authraquinones, terpenoids and organic acids. CONCLUSION: A method is established in this study to identify various chemical constituents of Naoxintong capsules rapidly, accurately and systematically. What's more, our research will lay a sound theoretical foundation and propose a scientific study idea for the quality control improvement, bioactive components recognition and further clinical application of this herbal formulation.

Study on chemical constituents of Tiandan Tongluo Capsule based on UPLC-Q-Orbitrap HRMS / 中草药

Objective: To clarify the chemical constituents of Tiandan Tongluo Capsule and establish a precise and effective identification method for the complex composition of TCM. Methods: This objective was achieved mainly depending on the information of the accurate mass and the multistage fragment ions obtained by ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS), comparing the relative retention time and the mass spectrometric data of the standard substance and consulting the reference literature. Results: Forty compounds were identified in this study, including the phenoliacids, anthraquinones, flavones, phthalides, fatty acids, and the others. Conclusion: This study can identify various chemical constituents of Tiandan Tongluo Capsule systematically, accurately, and rapidly. Moreover, the scientific theory basis will be provided for the pharmacodyamic material basis and the quality control of this drug at the same time.

Chemical constituents of Shenkun Yangxue Granules based on UPLC-Q-Orbitrap HRMS / 中草药

Objective To study the chemical compositions of Shenkun Yangxue Granules. A novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was established to rapidly and accurately recognize the complex chemical constituents of Chinese materia medica. Methods The information of accurate mass in Full-MS and multistage fragment ions was obtained by UPLC-Q-Orbitrap HRMS technology. Chemical constituents were identified by comparing their relative retention times and the mass data with those of reference substances, as well as consulting the reference literature or Chemical Book Network Database. Results Fifty-one compounds were finally identified from Shenkun Yangxue Granules, including the organic acids, flavones, quinones, glycosides, and others. Conclusion The method established in this study can identify various chemical constituents of Shenkun Yangxue Granules rapidly, accurately and systematically. What’s more, our research will lay a sound theoretical foundation and scientific study idea for improving the quality control, bioactive components recognition and further clinical application of this herbal formulation.

The multiple constituents study of Danhuang Quyu Capsule based on UPLC-Q-Orbitrap HRMS / 中草药

Objective In order to clarify the chemical constituents in traditional Chinese medicine (TCM) compound preparation of Danhuang Quyu Capsule and establish an efficient analysis method for the identification of the complex components in TCM accurately and rapidly. Methods An ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to perform an identification analysis of the chemical constituent in the Danhuang Quyu Capsule.This objective was achieved mainly depending on the information of the accurate mass and the multistage fragment ions obtained by UPLC-Q-Orbitrap, comparing the relative retention time and the massspectrometricdata of the standardsubstance and consulting the reference literature. Results Fifty nine compounds were identified in this study, including the flavones, phthalides, anthraquinones, alkaloids, monoterpene glycosides, organic acids, and other categories. Conclusion This study can identify various chemical constituents of Danhuang Quyu Capsule systematically, accurately and rapidly. What's more, the scientific theory basis will be provided for thepharmacodyamic material basis and the quality control of this drug at the same time. Key words: Danhuang Quyu Capsule; UPLC-Q-Orbitrap HRMS; flavones; phthalides; monoterpene glycosides