Fast screening of tyrosinase inhibitors from traditional Chinese medicinal plants by ligand fishing in combination with in situ fluorescent assay

A simple and rapid method for screening of tyrosinase (TYR) inhibitors present in traditional Chinese medicines (TCMs) was developed by combining ligand fishing and the fluorescent enzymatic assay based on dopamine-functionalized carbon quantum dots (CQDs-Dopa). Ligands of the enzyme present in the...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2022-03, Vol.414 (6), p.2265-2273
Main Authors: Zhao, Yan, Hu, Jin-Jie, Bai, Xiao-Lin, Liu, Hao-Peng, Qi, Xu-Wei, Liao, Xun
Format: Article
Language:English
Subjects:
Analysis
Analytical Chemistry
Assaying
Baicalin
Beads
Biochemistry
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
China
Chromatography, High Pressure Liquid - methods
Dopamine
Enzyme inhibitors
Enzymes
Fishing
Fluorescence
Food Science
Herbal medicine
Identification and classification
Laboratory Medicine
Ligands
Ligands (Biochemistry)
Liquid chromatography
Mass spectrometry
Mass spectroscopy
Medicinal plants
Medicine, Chinese
Methods
Monitoring/Environmental Analysis
Monophenol Monooxygenase
Oxidases
Plants, Medicinal
Properties
Quadrupoles
Quantum dots
Quercetin
Research Paper
Screening
Traditional Chinese medicine
Tyrosinase
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Summary:A simple and rapid method for screening of tyrosinase (TYR) inhibitors present in traditional Chinese medicines (TCMs) was developed by combining ligand fishing and the fluorescent enzymatic assay based on dopamine-functionalized carbon quantum dots (CQDs-Dopa). Ligands of the enzyme present in the TCM extractions were firstly adsorbed on the enzyme-modified magnetic beads, and then the beads were magnetically separated and subjected directly to the CQDs-Dopa-based fluorescent assay. Finally, compounds were desorbed from the “active” beads and identified with ultra-performance liquid chromatography-triple quadrupole mass spectrometry. A known natural TYR inhibitor quercetin was selected to assess the feasibility and quantification performance of this method, and good linearity in the range of 0.01–0.16 mM ( R 2  = 0.992) with a low detection limit of 0.004 mM was obtained. This method was then applied to screen TYR inhibitors present in Scutellaria baicalensis and Sophora flavescens . Six TYR inhibitors including baicalin ( 1 ), baicalein ( 2 ), wogonin ( 3 ), oroxylin A ( 4 ), kurarinone ( 5 ), and sophoraflavanone G ( 6 ) were found, among which 1 – 4 were firstly discovered in this work. This is the first report on the in situ assessment of the target compounds obtained by ligand fishing in the form of a mixture, which exhibited the combined advantages of specific extraction ability of ligand fishing and the high sensitivity of CQDs-based fluorescent assay, showing great potential for fast screening of enzyme inhibitors from TCMs. Graphical abstract
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-021-03864-w