Rapid identification of components in Wuzhuyu Decoction using UHPLC Q-Exactive Orbitrap MS~n and molecular network technology / 中国中药杂志

Wuzhuyu Decoction, the classical formula recorded in the Treatise on Febrile Diseases(Shang Han Lun), has been included in the Catalogue of Ancient Classic Prescriptions(the First Batch). Consisting of Euodiae Fructus, Ginseng Radix et Rhizoma, Zingiberis Rhizoma Recens, and Jujubae Fructus, it is effective in warming the middle, tonifying deficiency, dispelling cold, and descending adverse Qi, and is widely applied clinically with remarkable efficacies. For a classical formula, the chemical composition is the material basis and an important premise for quantity value transfer. This study aimed to establish a rapid identification method of chemical components in Wuzhuyu Decoction by high-resolution mass spectrometry(HR-MS) and molecular network. AQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was used for sample separation, and acetonitrile-0.1% formic acid in water was used as mobile phases for gradient elution. Q-Exactive Orbitrap MS data were collected in positive and negative ion modes, and GNPS molecular network was plotted according to the similarity of MS/MS fragmentation modes. Cytoscape 3.6.1 was used to screen molecular clusters with similar structures. Finally, the chemical components of Wuzhuyu Decoction were rapidly identified according to the controls, as well as the information of retention time, accurate relative molecular weight of HR-MS, and MS/MS multistage fragments. A total of 105 chemical components were identified in Wuzhuyu Decoction. This study can provide data for the follow-up quality control, standard substance research, and pharmacodynamic material research on Wuzhuyu Decoction, as well as references for the rapid qualitative analysis of the chemical components of Chinese medicine.

Comparison of alkaloids in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Praeparata based on UHPLC-Q-Exactive Orbitrap MS/MS / 中国中药杂志

UHPLC-Q-Exactive Orbitrap MS/MS was used to systematically analyze and compare the alkaloids in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Praeparata. After the samples were pretreated in the solid-phase extraction cartridges, 0.1% ammonium hydroxide(A)-acetonitrile(B) was used for gradient elution. The LC-MS method for characterization of alkaloids in the three herbal medicines was established in ESI positive ion mode to collect high resolution MS data of reference substances and samples. On the basis of the information of reference substance cracking behavior, retention time, accurate molecular mass, and related literature, a total of 155 alkaloids were identified in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Prae-parata. Specifically, 130, 127, and 92 alkaloids were identified in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Praeparata, respectively. Monoester alkaloids and amino-alcohol alkaloids were dominant in the three herbal medicines, and the alkaloids in Aconiti Kusnezoffii Radix and Aconiti Radix were similar. This paper can provide a reference for elucidating the pharmacological effects and clinical application differences of the three herbal medicines produced from plants of Aconitum.

Identification of Four Diterpenoid Alkaloids in Radix Aconiti Kusnezoffii by SCX-SPE Combined with UHPLC-Q-Exactive Orbitrap MS / 世界科学技术-中医药现代化

Objective To comprehensively characterize the diterpene alkaloids in Radix Aconiti Kusnezoffii.Methods The diterpene alkaloids were isolated and purified by strong acid cation exchange resin solid phase extraction column(SCX-SPE),and identified by ultra high performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry(UHPLC-Q-Exactive Orbitrap MS).Results A total of 99 diterpene alkaloids were identified from Radix Aconiti Kusnezoffii,including 27 diester diterpene alkaloids(DDA),29 monoester diterpene alkaloids(MDA),40 amide diterpene alkaloids(ADA),2 polyester diterpene alkaloids(PDA)and 1 long-chain ester diterpene alkaloid(LDA).Conclusion The SCX-SPE combined with UHPLC-Q-Exactive Orbitrap MS method,established in this paper,can rapidly identify a large number of diterpene alkaloids in Radix Aconiti Kusnezoffii,which provides scientific proof for the study of pharmacodynamic substance basis and quality control of Radix Aconiti Kusnezoffii.

Chemical analysis of classical prescription Qianghuo Shengshi Standard Decoction by UHPLC-Q Exactive Orbitrap MS / 中国中药杂志

A UHPLC-Q Exactive Orbitrap MS method was used to analyze the chemical constituents of the classical prescription Qianghuo Shengshi Standard Decoction(QHSS). UHPL conditions were as follows: Waters~(TM) UPLC~(TM) HSS T3 C_(18) column(2.1 mm×100 mm, 1.7 μm) and mobile phase of acetonitrile-0.1% formic acid aqueous solution. Mass spectrometry data of QHSS, each herb extract, and negative sample were collected in both positive and negative ion modes. The chemical constituents of QHSS were identified or tentatively identified based on the accurate molecular weight, retention time, MS fragmentation, comparison with reference substances, and literature reports. A total of 141 compounds were identified, including 18 amino acids, oligosaccharides, oligopeptides, and their derivatives, 19 phenolic acids, 44 coumarins, 18 flavonoids and chromones, 13 saponins, 17 phthalides, and 12 other components. This study comprehensively characterized the chemical constituents of QHSS, laying an experimental basis for the in-depth research on the material basis and quality control of QHSS.

UHPLC-Q-Exactive Orbitrap MS/MS-based rapid identification of chemical components in substance benchmark of Kaixin San / 中国中药杂志

Ultra-performance liquid chromatography-quadrupole-electrostatic field Orbitrap mass spectrometry(UHPLC-Q-Exactive Orbitrap MS/MS) was used for rapid identification of the chemical components in Kaixin San substance benchmark. The gradient elution was performed through a Waters ACQUITY~(TM) BEH C_(18) column(2.1 mm×150 mm, 1.7 μm) with water-acetonitrile as mobile phase, a column temperature of 30 ℃, a flow rate of 0.3 mL·min~(-1), and a sample size of 1 μL. The scanning was performed in the negative ion mode. The complex component groups in Kaixin San substance benchmark were quickly and accurately identified and clearly assigned based on the comparison of the retention time and MS data with those of the reference substance as well as the relative molecular weight of the same or similar components in the mass spectrum database and literature. A total of 77 compounds were identified, including 26 saponins, 13 triterpenoid acids, 20 oligosaccharide esters, 5 xanthones, and 13 other compounds. The qualitative method established in this study can systematically, accurately, and quickly identify the chemical components in Kaixin San substance benchmark, which can provide a basis for the further analysis of its active components in vivo and the establishment of its quality control system.

Study on effective components of Ganoderma applanation and its mechanism of action in preventing COVID-19 based on UHPLC-Q- Exactive-Orbitrap-MS integrated network pharmacology / 中国药理学通报

Aim To elucidate the effective components of Ganoderma applanatum and its mechanism of preventing the coronavirus disease 2019(COVID-19).Methods To begin with, UHPLC-Q-Exactive-Orbitrap-MS was established to identify the main chemical constituents of G.applanatum.Then, the predicted targets of G.applanatum were selected by Swiss Target Prediction.GO analysis and KEGG analysis of core target genes were performed using the DAVID database.Finally, to explore the potential mechanism of G.applanatum against COVID-19, core functional components-core target-metabolism path network diagram was constructed using Cytoscape 3.8.0, and molecular docking was used to analyze the binding force of the core effective compounds with angiotensin-converting enzyme II(ACE2)and three SARS CoV-2 proteins, nonstructural protein-15 Endoribonuclease(NSP15), the receptor-binding domain of spike protein(RBD of S protein), and main protease(Mpro/3CLpro).Results Sixty-two components were identified from G.applanatum by UHPLC-Q-Exactive-Orbitrap-MS study; 30 active components were closely associated with 32 core targets including IL6, PTGS2, and MAPK1; KEGG analysis showed that it might treat COVID-19 through signaling pathways, such as PI3K-Akt signaling pathway, TNF signaling pathway, tuberculosis, and so on; molecular docking analysis showed that 1,4-Dihydroxy-2-naphthoic acid, parthenolide, 7,8-Dihydroxycoumarin, and other vital compounds had a certain degree of affinity with ACE2 and three SARS CoV-2 proteins.Conclusion This study clarifies the chemical composition and the potential mechanism of G.applanatum, providing a scientific basis for screening the effective ingredients of G.applanatum.

Analysis of Metabolites and Metabolism Pathway of Tetrahydroxystilbene Glucoside in Rats with UHPLC-MS and Mass Defect Filter Method / 中国药房

OBJECTIVE：To analyze the metabolites of tetrahydroxystilbene glucoside （THSG）and speculate its metabolism pathway in rats. METHODS ：Male SD rats were randomly divided into plasma group （n＝3），urine group （n＝3），bile group （n＝3），and tissue group （n＝9）. Each group was given single dose of THSG 200 mg/kg intragastrically. Plasma samples 10，30 min and 1，1.5，2，4 h after medication ，the unrine 0-6 h after medication ，the bile 0-4 h after medication ，the tissue of heart ， liver，spleen，lung，kidney and stomach 30 min and 1，2 h after medication （3 at each time point ）were collected respectively.After precipitated with methanol ，the metabolites of samples were analyzed and identified by UHPLC-Q-Exactive Orbitrap MS and mass loss filtration （MDF）. Its metabolism pathway was speculated. RESULTS：In the blood ，urine，bile，heart，liver，spleen， lung，kidney，stomach samples ，6，7，11，1，5，1，3，4，4 metabolites were detected ，including two phase Ⅰ（hydrolysis， hydrogenation and hydroxylation ）metabolites，18 phase Ⅱ（glucuronic acid binding and sulfation ）metabolites. There were 12 glucuronic acid binding products. CONCLUSIONS：Most of the metabolites of THSG are found in bile ，mainly glucuronic acid binding products of phase Ⅱ metabolite THSG ； main metabolic pathways involve glucose hydrolysis ， hydrogenation， hydroxylation，glucuronic acid binding and sulfation.

Phospholipidomics study of Jiegeng Decotion for LPS-induced acute lung injury in mice based on UHPLC-Q-Exactive Orbitrap/MS / 药学学报

Using the lipidomics method based on UHPLC-Q-Exactive Orbitrap/MS, the change of phospholipid metabolism in lung tissue of mice induced by lipopolysaccharide (LPS)-induced acute lung injury was analyzed to observe the regulation of abnormal lipids by Jiegeng Decoction and to explore the regulation effect of Jiegeng Decoction on LPS-induced acute lung injury. The lung tissue samples from control group, model group, dexamethasone (positive drug) group, and Jiegeng Decoction group were collected and the lipid components of the sample were extracted. All procedures over mice were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of Nanjing University of Chinese Medicine, and the experiments were approved by the Animal Ethics Committee of our university. The lipidomics technique of UHPLC-Q-Exactive Orbitrap/MS was used to study change of phospholipids in lung tissue of each group. LPS induced acute lung injury in mice with metabolic abnormalities of phospholipids, the specific performance of the PC was significantly upregulated, phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), phosphatidyl serine (PS),phosphatidylinositol (PI) and other metabolic disorders, Jiegeng Decoction have a certain role in these phospholipids. LPS-induced acute lung injury caused disturbances of phospholipid in vivo, and Jiegeng Decoction regulates metabolic phospholipids.