Mass Spectrometry Techniques in Flavonoid Analysis

Mass spectrometry has become indispensable for elucidating the structural diversity, abundance and biological roles of flavonoids in plants and food products. Typically coupled with chromatographic separation—such as high-performance or ultra-high-performance liquid chromatography—modern mass spectrometers combine high resolution, accurate mass measurement and tandem fragmentation to distinguish isobaric and isomeric flavonoid species. Electrospray ionisation affords gentle ion generation in both positive and negative modes, enabling intact molecular ions and characteristic fragment ions to be recorded. Time-of-flight and orbitrap analysers deliver sub-ppm mass accuracy, allowing elemental composition assignments and metal-adduct profiling. Multistage MSn experiments reveal diagnostic neutral losses—such as sugar moieties or ring-cleavage fragments—thus clarifying glycosylation sites and aglycone substitution patterns. Advancements in chip-based nanospray, ion mobility separation and integration with theoretical modelling have refined interpretation of complex spectra and energy barriers in fragmentation pathways. These developments underpin applications ranging from chemotaxonomy and quality control of herbal medicines to metabolic profiling in ecological and nutritional studies, reinforcing the global significance of flavonoid research.

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Mass Spectrometry Techniques in Flavonoid Analysis publication trend

The graph below shows the total number of articles in mass spectrometry techniques in flavonoid analysis across all publications each year (not limited to Nature Index journals).

Technical terms

Electrospray ionisation (ESI): A soft ionisation technique that produces charged droplets of analyte suitable for mass analysis, allowing detection of intact flavonoid ions and their fragments.

Ultra-high-performance liquid chromatography (UHPLC): A chromatographic method using sub-2 µm particle columns for high-resolution separation of complex flavonoid mixtures prior to MS detection.

High-resolution mass spectrometry (HRMS): Analysers (e.g. orbitrap, time-of-flight) providing accurate mass measurements with sub-ppm precision to determine elemental compositions of flavonoid ions.

Tandem mass spectrometry (MS/MS or MSn): Sequential fragmentation of selected precursor ions to generate diagnostic product ions, enabling structural elucidation of glycosidic linkages and ring substituents.

Retro-Diels–Alder fragmentation: A concerted gas-phase cleavage of the C-ring in flavonoids yielding two complementary fragments, informative of ring connectivity and substitution patterns.

References

1. Metabolic profiling of Ochradenus baccatus Delile. utilizing UHPLC-HRESIMS in relation to the in vitro biological investigations. Food Chemistry (2023).
2. Structural Characterization of Flavonoid Glycoconjugates and Their Derivatives with Mass Spectrometric Techniques. Molecules (2016).
3. Chemotaxonomic Classification Applied to the Identification of Two Closely-Related Citrus TCMs Using UPLC-Q-TOF-MS-Based Metabolomics. Molecules (2017).
4. Characterisation of Flavonoid Aglycones by Negative Ion Chip‐Based Nanospray Tandem Mass Spectrometry. International Journal of Analytical Chemistry (2012).
5. The Mechanism of a Retro-Diels–Alder Fragmentation of Luteolin: Theoretical Studies Supported by Electrospray Ionization Tandem Mass Spectrometry Results. Molecules (2022).
6. The Use of Mass Spectrometric Techniques to Differentiate Isobaric and Isomeric Flavonoid Conjugates from Axyris amaranthoides. Molecules (2016).