Integrated UPLC-Q-TOF-MS and network pharmacology approach-driven quality marker discovery of Danggui Shaoyao San for primary dysmenorrhea.

Danggui Shaoyao San (DSS), a famous prescription, has been clinically proved to be effective in treating primary dysmenorrhea (PD). Currently there is no valid quality control data available for DSS. The main aim of the current research was to explore quality markers (Q-markers) of DSS. The chemical constituents of DSS were qualitatively identified using ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) technology. On this basis, the targets of DSS and PD were predicted and screened using the TCMSP, SwissTargetPrediction, GeneCards, OMIM and TTD databases. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis was performed on the core intersection targets using string and Cytoscape 3.7.1 software. Then molecular docking was conducted to screen the Q-markers of DSS in PD. A total of 126 chemical constituents, including 22 organic acids, 14 phthalides, 24 monoterpenoids, five sesquiterpene lactones, 22 triterpenoids, four phenylpropanoids and 35 other compounds were preliminarily characterized. According to network pharmacology prediction analysis, six compounds containing polyporenic acid C, senkyunolide P, alisol B 23-acetate, naringenin, gallic acid, ferulic acid and albiflorin were regarded as Q-markers of DSS. The present research established an integrative UPLC-Q-TOF-MS and network pharmacology method to discover the latent Q-markers of DSS and provided a theoretical data for the follow-up quality control of DSS.

Comprehensive Characterization and Identification of Chemical Constituents of Xiangsha Pingwei Pills by UPLC-Q-TOF-MS.

BACKGROUND: Xiangsha Pingwei Pills (XPP) is a traditional Chinese medicine (TCM) prescription, which is widely used to treat epigastric pain in China. Its systematic chemical characteristics have rarely been reported, which hinders the interpretation of the material basis of its prescription. OBJECTIVE: To establish a rapid and effective component characterization method for XPP using ultra-HPLC-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) and the data post-processing program, Peakview 1.2 software. METHODS: A UPLC-Q-TOF-MS method coupled with Peakview 1.2 software was successfully established for the first time to investigate the complex constituents of XPP. Accurate MS and MS/MS data were detected in positive and negative ion mode. The compounds were tentatively identified based on their retention times, MS, and MS/MS data, as well as reference standards and from the literature. RESULTS: The chemical profile of XPP was acquired, and a total of 130 compounds in XPP were preliminarily identified for the first time, including 6 organic acids, 59 flavonoids, 13 lignans, 20 terpenoids, 9 phenylpropanoids, 6 alkaloids, 5 amino acids, and 12 other compounds. CONCLUSION: A rapid and effective UPLC-Q-TOF-MS method for the main chemical components of XPP has been established for further characterizing constituents and the overall quality control of XPP. HIGHLIGHTS: This is the first report of a comprehensive analysis method for the main chemical components of XPP, which aims to lay a solid foundation for the chemical basis and overall quality control of XPP.

Uncovering the effects and mechanism of Danggui Shaoyao San intervention on primary dysmenorrhea by serum metabolomics approach.

Danggui Shaoyao San (DSS) is a well-known prescription for relieving primary dysmenorrhea (PD) of women in China. However, its pharmacological mechanism has not been thoroughly uncovered. Here, an integrative UPLC-Q-TOF-MS-based serum metabolomics approach coupled with multivariate data analysis has been proposed to investigate the effects and mechanism of DSS on estradiol benzoate and oxytocin-induced PD rats. 31 potential biomarkers of PD were screened and identified, mainly involving phenylalanine, tyrosine and tryptophan biosynthesis, glycerophospholipid metabolism, primary bile acid biosynthesis, and the occurrence of PD could destroy biological homeostasis in vivo by monitoring these pathways. After DSS treatment, 18 identified different metabolites were restored to the nomal state in varying degrees and could be potential biomarkers contributing to the treatment of DSS. These findings implyed that DSS exhibited a therapeutic effect on PD rats through regulating multiple abnormal pathways. Of note, this study discovered some potential biomarkers related to PD for the first time, such as L-tyrosine, glycocholic acid, citric acid, palmitoylcarnitine, cholesterol. It preliminarily proved the pathophysiology of PD and action mechanisms of DSS on PD, and provided a novel insight into the effectiveness of DSS on PD.

Integrated Serum Pharmacochemistry, Metabolomics, and Network Pharmacology to Reveal the Material Basis and Mechanism of Danggui Shaoyao San in the Treatment of Primary Dysmenorrhea.

Danggui Shaoyao San (DSS), a well-known formula, has been successfully applied in treating primary dysmenorrhea (PD) in China. However, its material basis and mechanism are still unrevealed. This current research aims to reveal the material basis and mechanism of DSS in treating PD by an integrative approach of serum pharmacochemistry, metabolomics, and network pharmacology. The results showed that DSS markedly relieved the physiological and pathological symptoms of PD as confirmed by the improvement of writhing behavior, inhibition of uterine edema, callback of clinical biochemical indexes, and metabolic profiles. Furthermore, a metabolomic analysis demonstrated that the therapeutic effect of DSS was attributed to the modulation of arachidonic acid metabolism, pentose and glucuronate interconversions, and phenylalanine metabolism. Meanwhile, 23 blood ingredients were identified after the oral administration of DSS. By analyzing the correlation coefficient of the identified biomarkers and blood components, active compounds closely associated with core metabolic pathways were extracted. Taking these active compounds as a basis, network pharmacology prediction was executed. It was found that active components of DSS including alisol B,23-acetate, chlorogenic acid, levistilide A, cianidanol, senkyunolide A, atractylenolide II, and sedanolide, were germane to steroid hormone biosynthesis, arachidonic acid metabolism, sphingolipid signaling pathway, etc. Interestingly, PTGS2 and PTGS1 related to the arachidonic acid metabolism may be pivotal targets of DSS. The current study proved that the integration of serum pharmacochemistry, metabolomics, and network pharmacology, was a powerful approach to investigate the material basis and the molecular mechanisms of DSS, and provided a solid basis for DSS application.