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标题:Functionalized magnetic nanoparticles coupled with mass spectrometry for screening and identification of cyclooxygenase-1 inhibitors from natural products.
时间:2020-03-27 00:02:12
DOI:10.1016/j.jchromb.2014.04.032
PMID:24793085
作者:Zhang, Yuping;Shi, Shuyun;Chen, Xiaoqin
关键词:Cyclooxygenase-1; Ligand fishing; MS; Turmeric; Curcuminoid
出版源: 《Journal of Chromatography B》 ,960 :126-132
摘要:Development of simple and effective methods for high-throughput, high-fidelity screening and identification of cyclooxygenase-1 (COX-1) inhibitors from natural products are important for drug discovery to treat inflammation and carcinogenesis. Here, we developed a new screening assay based on cyclooxygenase-1 (COX-1) functionalized magnetic nanoparticles (i.e. Fe3O4@SiO2-COX-1) for solid phase ligand fishing, and then mass spectrometry (MS) was applied for structural identification. Incubation conditions were optimized. High specificity for isolating COX-1 inhibitors was achieved by testing positive control, indomethacin, with active and inactive COX-1. Moreover, high stability of immobilized COX-1 (remained 95.3% after ten consecutive cycles) allows the analysis reproducible. When applied to turmeric extract, four curcuminoids (i.e. curcumin, demethoxycurcumin, bisdemethoxycurcumin, and 1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)-(1E,6E)-1,6-heptadiene-3,5-dione), difficult to be distinguished from original MS spectrum of turmeric extract, were isolated as main COX-1 inhibitors. Their structures were characterized based on their accurate molecular weight and diagnostic fragment ions. The results indicated that the proposed method was a simple, robust and reproducible approach for the discovery of COX-1 inhibitors from complex matrixes. (C) 2014 Elsevier B.V. All rights reserved.
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目录:
  • Functionalized magnetic nanoparticles coupled with mass spectrometry for screening and identification of cyclooxygenase-1 ...
    • 1 Introduction
    • 2 Experimental
      • 2.1 Chemicals and reagents
      • 2.2 Preparation of turmeric extraction
      • 2.3 COX-1 inhibition assay
      • 2.4 COX-1 functionalized magnetic nanoparticles for ligand fishing
        • 2.4.1 Preparation of Fe3O4@SiO2–COX-1 nanoparticles
        • 2.4.2 Characterizations of Fe3O4@SiO2–COX-1 nanoparticles
        • 2.4.3 Assay verification
        • 2.4.4 Ligand fishing assay
      • 2.5 HPLC-DAD and ESI-MS analysis
    • 3 Results and discussion
      • 3.1 Characterization of Fe3O4@SiO2–COX-1 nanoparticles
      • 3.2 Assay verification
      • 3.3 Reusability and reproducibility of immobilized COX-1
      • 3.4 Analysis of curcuminoids by ESI-MS
      • 3.5 COX-1 inhibitors in turmeric extract
    • 4 Conclusion
    • Acknowledgements
    • Appendix A Supplementary data
    • Appendix A Supplementary data

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