Objective: The objective of this study was to decipher chemical interactions between Danshen and Danggui using liquid chromatography–mass spectrometry(LC-MS) and explore the mechanisms of Danshen–Danggui against str...Objective: The objective of this study was to decipher chemical interactions between Danshen and Danggui using liquid chromatography–mass spectrometry(LC-MS) and explore the mechanisms of Danshen–Danggui against stroke using network pharmacology and molecular docking. Materials and Methods: First, the chemical compounds of Danshen–Danggui were profiled using ultra-high-performance liquid chromatography(HPLC)-quadrupole time-of-flight MS. Accurately characterized compounds in various proportions of Danshen–Danggui were quantified using HPLC combined with triple quadrupole electrospray tandem MS. Network pharmacology was used to uncover the essential mechanisms of action of Danshen–Danggui against stroke. Discovery Studio Software was used for the molecular docking verification of key active chemicals and stroke-related targets. Results: A total of 53 compounds were characterized, and 22 accurately identified constituents(10 phenolic acids, 8 phthalides, and 4 tanshinones) were quantified in 15 proportions of Danshen–Danggui. The quantification results showed that Danggui significantly increased the dissolution of most phenolic acids(compounds from Danshen), whereas Danshen promoted the dissolution of most phthalides(compounds from Danggui). Overall, the combination of Danshen and Danggui at a 1:1 ratio resulted in the maximum total dissolution rate. Further network pharmacology and molecular docking results indicated that Danshen–Danggui exerted anti-stroke effects mainly by regulating inflammation-related(tumor necrosis factor, hypoxia-inducible factor, and toll-like receptor) signaling pathways, which ranked among the top three pathways based on Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. Conclusion: The chemical compounds in Danshen–Danggui could interact with each other to increase the dissolution of the most active compounds, which could provide a solid basis for uncovering the compatibility mechanisms of Danshen–Danggui and Danshen–Danggui-based formulae.展开更多
RNA modification has recently been proposed to play important roles in biological regulation. The detection and quantification of RNA modifications generally are challenging tasks since most of the modifications exist...RNA modification has recently been proposed to play important roles in biological regulation. The detection and quantification of RNA modifications generally are challenging tasks since most of the modifications exist in low abundance in vivo. Here we developed an on-line trapping/capillary hydrophilic-interaction liquid chromatography/electrospray ionization-mass spectrometry(on-line trapping/cHILIC/MS) method for sensitive and simultaneous quantification of RNA modifications of N^6-methyladenosine(m^6A) and 5-methylcytosine(5-mC) from human blood. The hydrophilic organic-silica hybrid monolith was prepared using sol-gel combined with "thiol-ene" click reaction for the separation of nucleosides. A poly(MAA-co-EGDMA) monolithic capillary was used as the on-line trapping column.With the developed on-line trapping/cHILIC/MS analytical platform, the detection limits of m^6A and 5-mC can reach to 0.06 fmol and 0.10 fmol. We then investigated the contents of m^6A and 5-mC in human blood RNA from healthy persons at the age of 6-14 and 60-68 years. Our results showed that both m^6A and 5-mC contents were significantly decreased in elder persons, suggesting the RNA modifications of m^6A and 5-mC are correlated to aging.展开更多
Metabolite analysis or metabolomics is an important component of systems biology in the post-genomic era.Although separate liquid chromatography（LC） methods for quantification of the major classes of polar metabolit...Metabolite analysis or metabolomics is an important component of systems biology in the post-genomic era.Although separate liquid chromatography（LC） methods for quantification of the major classes of polar metabolites of plants have been available for decades,a single method that enables simultaneous determination of hundreds of polar metabolites is possible only with gas chromatography-mass spectrometry（GC-MS） techniques.The rapid expansion of new LC stationary phases in the market and the ready access of mass spectrometry in many laboratories provides an excellent opportunity for developing LC-MS based methods for multitarget quantification of polar metabolites.Although various LC-MS methods have been developed over the last 10 years with the aim to quantify one or more classes of polar compounds in different matrices,currently there is no consensus LC-MS method that is widely used in plant metabolomics studies.The most promising methods applicable to plant metabolite analysis will be reviewed in this paper and the major problems encountered highlighted.The aim of this review is to provide plant scientists,with limited to moderate experience in analytical chemistry,with up-to-date and simplified information regarding the current status of polar metabolite analysis using LC-MS techniques.展开更多
As the roles of glycans in health and disease continue to be unraveled,it is becoming apparent that glycans’immense complexity cannot be ignored.To fully delineate glycan structures,we developed an integrative approa...As the roles of glycans in health and disease continue to be unraveled,it is becoming apparent that glycans’immense complexity cannot be ignored.To fully delineate glycan structures,we developed an integrative approach combining a set of cost-effective,widespread,and easy-to-handle analytical methods.The key feature of our workflow is the exploitation of a removable fluorescent label—exemplified by 9-fluorenylmethyl chloroformate(Fmoc)—to bridge the gap between diverse glycoanalytical methods,especially multiplexed capillary gel electrophoresis with laser-induced fluorescence detection(xCGELIF)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOFMS).Through the detailed structural analysis of selected,dauntingly complex N-glycans from chicken ovalbumin,horse serum,and bovine transferrin,we illustrate the capabilities of the presented strategy.Moreover,this approach“visualizes”N-glycans that have been difficult to identify thus far—such as the sulfated glycans on human immunoglobulin A—including minute changes in glycan structures,potentially providing useful new targets for biomarker discovery.展开更多
基金funded by the State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources (Guangxi Normal University) (CMEMR2022-B11)National Natural Science Foundation of China (822044593)Natural Science Foundation of Jiangsu Higher Education Institutions of China (22KJB360018)。
文摘Objective: The objective of this study was to decipher chemical interactions between Danshen and Danggui using liquid chromatography–mass spectrometry(LC-MS) and explore the mechanisms of Danshen–Danggui against stroke using network pharmacology and molecular docking. Materials and Methods: First, the chemical compounds of Danshen–Danggui were profiled using ultra-high-performance liquid chromatography(HPLC)-quadrupole time-of-flight MS. Accurately characterized compounds in various proportions of Danshen–Danggui were quantified using HPLC combined with triple quadrupole electrospray tandem MS. Network pharmacology was used to uncover the essential mechanisms of action of Danshen–Danggui against stroke. Discovery Studio Software was used for the molecular docking verification of key active chemicals and stroke-related targets. Results: A total of 53 compounds were characterized, and 22 accurately identified constituents(10 phenolic acids, 8 phthalides, and 4 tanshinones) were quantified in 15 proportions of Danshen–Danggui. The quantification results showed that Danggui significantly increased the dissolution of most phenolic acids(compounds from Danshen), whereas Danshen promoted the dissolution of most phthalides(compounds from Danggui). Overall, the combination of Danshen and Danggui at a 1:1 ratio resulted in the maximum total dissolution rate. Further network pharmacology and molecular docking results indicated that Danshen–Danggui exerted anti-stroke effects mainly by regulating inflammation-related(tumor necrosis factor, hypoxia-inducible factor, and toll-like receptor) signaling pathways, which ranked among the top three pathways based on Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. Conclusion: The chemical compounds in Danshen–Danggui could interact with each other to increase the dissolution of the most active compounds, which could provide a solid basis for uncovering the compatibility mechanisms of Danshen–Danggui and Danshen–Danggui-based formulae.
基金financially supported by the National Natural Science Foundation of China (Nos. 21522507, 21672166, 21728802, 21721005)
文摘RNA modification has recently been proposed to play important roles in biological regulation. The detection and quantification of RNA modifications generally are challenging tasks since most of the modifications exist in low abundance in vivo. Here we developed an on-line trapping/capillary hydrophilic-interaction liquid chromatography/electrospray ionization-mass spectrometry(on-line trapping/cHILIC/MS) method for sensitive and simultaneous quantification of RNA modifications of N^6-methyladenosine(m^6A) and 5-methylcytosine(5-mC) from human blood. The hydrophilic organic-silica hybrid monolith was prepared using sol-gel combined with "thiol-ene" click reaction for the separation of nucleosides. A poly(MAA-co-EGDMA) monolithic capillary was used as the on-line trapping column.With the developed on-line trapping/cHILIC/MS analytical platform, the detection limits of m^6A and 5-mC can reach to 0.06 fmol and 0.10 fmol. We then investigated the contents of m^6A and 5-mC in human blood RNA from healthy persons at the age of 6-14 and 60-68 years. Our results showed that both m^6A and 5-mC contents were significantly decreased in elder persons, suggesting the RNA modifications of m^6A and 5-mC are correlated to aging.
基金funded by the Dairy Futures Co-operative Research Centre
文摘Metabolite analysis or metabolomics is an important component of systems biology in the post-genomic era.Although separate liquid chromatography（LC） methods for quantification of the major classes of polar metabolites of plants have been available for decades,a single method that enables simultaneous determination of hundreds of polar metabolites is possible only with gas chromatography-mass spectrometry（GC-MS） techniques.The rapid expansion of new LC stationary phases in the market and the ready access of mass spectrometry in many laboratories provides an excellent opportunity for developing LC-MS based methods for multitarget quantification of polar metabolites.Although various LC-MS methods have been developed over the last 10 years with the aim to quantify one or more classes of polar compounds in different matrices,currently there is no consensus LC-MS method that is widely used in plant metabolomics studies.The most promising methods applicable to plant metabolite analysis will be reviewed in this paper and the major problems encountered highlighted.The aim of this review is to provide plant scientists,with limited to moderate experience in analytical chemistry,with up-to-date and simplified information regarding the current status of polar metabolite analysis using LC-MS techniques.
基金support from the German Federal Ministry of Education and Research(BMBF)under the project“Die Golgi Glykan Fabrik 2.0”(031A557C for Samanta Cajic and Erdmann Rapp)the European Commission(EC)under the project“HighGlycan”(278535 for RenéHennig and Erdmann Rapp)the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)under the project“The concert of dolicholbased glycosylation:from molecules to disease models”(FOR2509 for Valerian Grote and Erdmann Rapp).
文摘As the roles of glycans in health and disease continue to be unraveled,it is becoming apparent that glycans’immense complexity cannot be ignored.To fully delineate glycan structures,we developed an integrative approach combining a set of cost-effective,widespread,and easy-to-handle analytical methods.The key feature of our workflow is the exploitation of a removable fluorescent label—exemplified by 9-fluorenylmethyl chloroformate(Fmoc)—to bridge the gap between diverse glycoanalytical methods,especially multiplexed capillary gel electrophoresis with laser-induced fluorescence detection(xCGELIF)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOFMS).Through the detailed structural analysis of selected,dauntingly complex N-glycans from chicken ovalbumin,horse serum,and bovine transferrin,we illustrate the capabilities of the presented strategy.Moreover,this approach“visualizes”N-glycans that have been difficult to identify thus far—such as the sulfated glycans on human immunoglobulin A—including minute changes in glycan structures,potentially providing useful new targets for biomarker discovery.
