An integrated strategy for the identification and screening of anti-allergy components from natural products based on calcium fluctuations and cell extraction coupled with HPLC–Q–TOF–MS

AbstractExtracts of Cyclocarya paliurus (CP) leaves, a popular sweet tea, inhibit pancreatic β cell apoptosis and have potent hypoglycemic effects, but the identities of the anti-apoptotic bioactive components are still unknown. In the present study, a method using UPLC-Q-TOF/MS based on serum pharmacochemistry combined with target cell extraction was established to rapidly identify direct-acting pancreatic protectants from CP. After orally administering a set amount of CP extract to rats, blood samples were collected to characterize the components that can be absorbed into the blood using UPLC-Q-TOF/MS. Also, target cells (pancreatic β NIT-1 cells) were incubated with CP extract for 24 hours, and cells were collected to identify the components that can bind to the cells using UPLC-Q-TOF/MS. Finally, to evaluate the protective effect of the bioactive components of CP, MTT and TUNEL assays were performed on treated NIT-1 cell induced by streptozotocin (STZ). Three potential direct-acting pancreatic protectants -- kaempferol, quercetin, quadranoside IV -- were identified, and anti-apoptotic effects of kaempferol and quercetin were confirmed in STZ-induced NIT-1 cells. The findings indicate that this combined approach is a feasible, rapid, and expedient tool for capturing potential direct-acting components from natural products such as those from CP leaves.

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Mori Ramulus (Chin.Ph.)—the Dried Twigs of Morus alba L./Part 1: Discovery of Two Novel Coumarin Glycosides from the Anti-Hyperuricemic Ethanol Extract

Molecules ◽

10.3390/molecules24030629 ◽

2019 ◽

Vol 24 (3) ◽

pp. 629

Author(s):

Jianbiao Yao ◽

Houhong He ◽

Jin Xue ◽

Jianfang Wang ◽

Huihui Jin ◽

...

Keyword(s):

Natural Products ◽

Ethanol Extract ◽

Accurate Method ◽

Morus Alba ◽

Strongly Correlated ◽

Bioactive Constituents ◽

Antiplatelet Aggregation ◽

Pharmacologically Active ◽

First Time ◽

Tof Ms

In Traditional Chinese Medicine (TCM), Mori ramulus (Chin.Ph.)—the dried twigs of Morus alba L.—is extensively used as an antirheumatic agent and also finds additional use in asthma therapy. As a pathological high xanthine oxidase (XO, EC 1.1.3.22) activity is strongly correlated to hyperuricemy and gout, standard anti-hyperuremic therapy typically involves XO inhibitors like allopurinol, which often cause adverse effects by inhibiting other enzymes involved in purine metabolism. Mori ramulus may therefore be a promissing source for the development of new antirheumatic therapeutics with less side effects. Coumarins, one of the dominant groups of bioactive constituents of M. alba, have been demonstrated to possess anti-inflammatory, antiplatelet aggregation, antitumor, and acetylcholinesterase (AChE) inhibitory activities. The combination of HPLC (DAD) and Q-TOF technique could give excellent separating and good structural characterization abilities which make it suitable to analyze complex multi-herbal extracts in TCM. The aim of this study was to develop a HPLC (DAD)/ESI-Q-TOF-MS/MS method for the identification and profiling of pharmacologically active coumarin glycosides in Mori ramulus refined extracts for used in TCM. This HPLC (DAD)/ESI-Q-TOF-MS/MS method provided a rapid and accurate method for identification of coumarin glycosides—including new natural products described here for the first time—in the crude extract of M. alba L. In the course of this project, two novel natural products moriramulosid A (umbelliferone-6-β-d-apiofuranosyl-(1→6)-β-d-glucopyranoside) and moriramulosid B (6-[[6-O-(6-deoxy-α-l-mannopyranosyl)-β-d-glucopyranosyl]oxy]-2H-1-benzopyran-1-one) were newly discovered and the known natural product Scopolin was identified in M. alba L. for the first time.

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An integrated strategy to improve data acquisition and metabolite identification by time-staggered ion lists in UHPLC/Q-TOF MS-based metabolomics

Journal of Pharmaceutical and Biomedical Analysis ◽

10.1016/j.jpba.2018.05.020 ◽

2018 ◽

Vol 157 ◽

pp. 171-179 ◽

Cited By ~ 9

Author(s):

Yang Wang ◽

Ruibing Feng ◽

Chengwei He ◽

Huanxing Su ◽

Huan Ma ◽

...

Keyword(s):

Data Acquisition ◽

Metabolite Identification ◽

Integrated Strategy ◽

Tof Ms

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Powerful GC-TOF-MS Techniques for Screening, Identification and Quantification of Halogenated Natural Products

Mass Spectrometry ◽

10.5702/massspectrometry.s0018 ◽

2013 ◽

Vol 2 (Special_Issue) ◽

pp. S0018-S0018 ◽

Cited By ~ 9

Author(s):

Peter S. Haglund ◽

Karin Löfstrand ◽

Kevin Siek ◽

Lillemor Asplund

Keyword(s):

Natural Products ◽

Halogenated Natural Products ◽

Tof Ms ◽

Identification And Quantification

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Identification of Photorhabdus symbionts by MALDI-TOF mass spectrometry

10.1101/2020.01.10.901900 ◽

2020 ◽

Author(s):

Virginia Hill ◽

Peter Kuhnert ◽

Matthias Erb ◽

Ricardo A. R. Machado

Keyword(s):

Mass Spectrometry ◽

Natural Products ◽

Related Species ◽

Entomopathogenic Nematodes ◽

Species Level ◽

Closely Related Species ◽

Maldi Tof Ms ◽

Bacterial Genus ◽

Maldi Tof ◽

Tof Ms

AbstractSpecies of the bacterial genus Photorhabus live in a symbiotic relationship with Heterorhabditis entomopathogenic nematodes. Besides their use as biological control agents against agricultural pests, some Photorhabdus species are also a source of natural products and are of medical interest due to their ability to cause tissue infections and subcutaneous lesions in humans. Given the diversity of Photorhabdus species, rapid and reliable methods to resolve this genus to the species level are needed. In this study, we evaluated the potential of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of Photorhabdus species. To this end, we established a collection of 54 isolates consisting of type strains and multiple field strains that belong to each of the validly described species and subspecies of this genus. Reference spectra for the strains were generated and used to complement a currently available database. The extended reference database was then used for identification based on the direct transfer sample preparation method and protein fingerprint of single colonies. High discrimination of distantly related species was observed. However, lower discrimination was observed with some of the most closely related species and subspecies. Our results, therefore, suggest that MALDI-TOF MS can be used to correctly identify Photorhabdus strains at the genus and species level, but has limited resolution power for closely related species and subspecies. Our study demonstrates the suitability and limitations of MALDI-TOF-based identification methods for the assessment of the taxonomical position and identification of Photorhabdus isolates.Impact StatementSpecies of the bacterial genus Photorhabus live in close association with soil-born entomopathogenic nematodes. Under natural conditions, these bacteria are often observed infecting soil-associated arthropods, but under certain circumstances, can also infect humans. They produce a large variety of natural products including antibiotics, insecticides, and polyketide pigments that have substantial agricultural, biotechnological and medical potential. In this study, we implement MALDI-TOF MS-based identification method to resolve the taxonomic identity of this bacterial genus, providing thereby a rapid identification tool to understanding its taxonomic diversity to boost scientific progress in medical, agricultural, and biotechnological settings.

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Integrated Strategy of UHPLC-Q-TOF-MS and Molecular Networking for Identification of Diterpenoids from Euphorbia fischeriana Steud. and Prediction of the Anti-Breast-Cancer Mechanism by the Network Pharmacological Method

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/3829434 ◽

2021 ◽

Vol 2021 ◽

pp. 1-19

Author(s):

Tian-Cheng Ma ◽

Yu-Kun Ma ◽

Jin-Ling Zhang ◽

Lei Liu ◽

Jia Sun ◽

...

Keyword(s):

Breast Cancer ◽

Signaling Pathway ◽

Complementary Therapy ◽

Mapk Signaling ◽

Pathway Enrichment Analysis ◽

Molecular Networking ◽

Docking Simulations ◽

Integrated Strategy ◽

Euphorbia Fischeriana ◽

Tof Ms

Breast cancer is one of the most common malignancies in women worldwide. Traditional Chinese medicine has been used as adjunctive or complementary therapy for breast cancer. Diterpenoids from Euphorbia fischeriana Steud. have been demonstrated to possess anti-breast-cancer activity. This research was aimed to systematically explore the diterpenoids from E. fischeriana and study the multiple mechanisms on breast cancer. The structures of diterpenoids were identified by the integrated strategy of UHPLC-Q-TOF-MS and molecular networking. A total of 177 diterpenoids belonging to 13 types were collected. In silico ADME analysis was performed on these compounds. It indicated that 130 of 177 diterpenoids completely adjusted to Lipinski’s rule. The targets of compounds were obtained from PharmMapper. The targets of breast cancer were collected from GeneCards. Then, 197 compounds-related targets and 544 breast cancer-related targets were identified. After the intersection process, 58 overlapping targets between compounds-related targets and breast cancer-related targets were acquired. The STRING database was applied to predict the protein-protein interactions. The GO and KEGG pathway enrichment analysis were performed by using the KOBAS database. It indicated that these predicted pathways were closely related to breast cancer. The treatment effect of E. fischeriana on breast cancer might be performed through signaling pathways, such as IL-17 signaling pathway, MAPK signaling pathway, and PI3K-Akt signaling pathway. The predicted top genes such as EGFR, ESR, MAPK, SRC, CASP3, CDK2, and KDR were involved in cell proliferation, gene transcription, apoptosis, signal transduction, DNA damage and repair, tumor differentiation, metastasis, and cell cycle, which indicated that E. fischeriana might treat breast cancer comprehensively. A compounds-KEGG pathways-related targets network was built by using cytoHubba to analyze the hub compounds and targets. It concluded that E. fischeriana treated breast cancer not only by the main components but also by the microconstituents, which reflected the overall regulatory role of multicomponents treating breast cancer. To estimate the binding affinities, binding sites, and binding postures, molecular docking simulations between 177 diterpenoids and top 19 targets were carried out. The results are basically in line with expectations. In conclusion, these results can serve as references for researchers studying potential targets of diterpenoids from E. fischeriana on breast cancer in the future.

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Generation of a Collision Cross Section Library for Multi-Dimensional Plant Metabolomics Using UHPLC-Trapped Ion Mobility-MS/MS

Metabolites ◽

10.3390/metabo10010013 ◽

2019 ◽

Vol 10 (1) ◽

pp. 13 ◽

Cited By ~ 4

Author(s):

Mark Schroeder ◽

Sven W. Meyer ◽

Heino M. Heyman ◽

Aiko Barsch ◽

Lloyd W. Sumner

Keyword(s):

Natural Products ◽

Cross Section ◽

Ion Mobility ◽

Collision Cross Section ◽

Molecular Formula ◽

Trapped Ion ◽

Grand Challenges ◽

Plant Natural Products ◽

Data Matching ◽

Tof Ms

The utility of metabolomics is well documented; however, its full scientific promise has not yet been realized due to multiple technical challenges. These grand challenges include accurate chemical identification of all observable metabolites and the limiting depth-of-coverage of current metabolomics methods. Here, we report a combinatorial solution to aid in both grand challenges using UHPLC-trapped ion mobility spectrometry coupled to tandem mass spectrometry (UHPLC-TIMS-TOF-MS). TIMS offers additional depth-of-coverage through increased peak capacities realized with the multi-dimensional UHPLC-TIMS separations. Metabolite identification confidence is simultaneously enhanced by incorporating orthogonal collision cross section (CCS) data matching. To facilitate metabolite identifications, we created a CCS library of 146 plant natural products. This library was generated using TIMS with N2 drift gas to record the TIMSCCSN2 of plant natural products with a high degree of reproducibility; i.e., average RSD = 0.10%. The robustness of TIMSCCSN2 data matching was tested using authentic standards spiked into complex plant extracts, and the precision of CCS measurements were determined to be independent of matrix affects. The utility of the UHPLC-TIMS-TOF-MS/MS in metabolomics was then demonstrated using extracts from the model legume Medicago truncatula and metabolites were confidently identified based on retention time, accurate mass, molecular formula, and CCS.

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