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Isolation, separation, identification, and quantification of bioactive methylated flavone regioisomers by UHPLC-MS/MS

Abstract:
Methylated flavones, commonly found in many plants of the Brassicaceae family, have potent antioxidant and anticancer activity with diverse therapeutic potential. However, the specific regioisomers of methylated flavones can have significantly different biochemical and potentially therapeutic properties as shown by various bioassays but analytically differentiating these compounds has been technically challenging and rarely reported. In this study, we demonstrate differentiation and identification of selected bioactive methylated flavone regioisomers, namely 5,7,3′-trihydroxy-4′-methoxyflavone, and 5,7,4′-trihydroxy-3′-methoxyflavone extracted from Coronopus didymus, a member of the Brassicaceae family, using ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-QTOF-MS/MS). Characteristic MS/MS product ions produced from neutral loss of carbon monoxide, and a methyl radical from the [M-H]– ion, exhibited differential relative abundances attributed to different structural stabilities under the same activation and collision-induced dissociation conditions. MS/MS also provided structural information which was sufficient to differentiate the methylated regioisomers and determine the position of the methyl group based on interpretation of their respective fragmentation patterns. Quantification showed 5,7,4′-trihydroxy-3′-methoxyflavone was at least 1.60 mg per 10 g plant material in C. didymus extracts. This study demonstrates a straightforward and novel approach to rapidly differentiate, identify and quantify regio-isomeric methylated flavone natural products using reversed-phase UPLC-MS/MS.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1002/ansa.202100016

Authors


More by this author
Institution:
University of Oxford
Department:
Plant Sciences
Sub department:
Biochemistry
Oxford college:
Magdalen College
Role:
Author
ORCID:
0000-0002-6628-323X
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Chemistry
Oxford college:
Merton College
Role:
Author
ORCID:
0000-0003-4733-1205


Publisher:
Wiley
Journal:
Analytical Science Advances More from this journal
Volume:
2
Issue:
7-8
Pages:
364-372
Publication date:
2021-05-05
Acceptance date:
2021-04-15
DOI:
EISSN:
2628-5452


Language:
English
Keywords:
Pubs id:
1176800
Local pid:
pubs:1176800
Deposit date:
2021-07-06

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