Mass spectrometry is crucial for analysing physicochemical and sensory properties, including colour, astringency, taste, and flavour, predicting ageing characteristics, and addressing stability issues in wine. Polyphenols are key chemical constituents in wine that are associated with health benefits and improve circulatory conditions. Advances in mass spectrometry ionisation techniques such as matrix-assisted laser desorption and ionisation and direct analysis in real-time offer high sensitivity for identifying important polyphenolic constituents in wine. High-resolution mass spectrometry, in combination with liquid chromatography, accurately identify and quantify polyphenolic compounds, even at low concentrations, and provides the possibility for further retrospective analysis and non-targeted analysis using statistical methods of data analysis. Ambient mass spectrometry techniques such as paper spray and low-temperature plasma allow solventless analysis, determining the geographical origin, authentication, and quality control of wine samples. This review will explore the potential benefits of using mass spectrometry to identify various polyphenols and polymeric polyphenols in wine, as well as recent developments and applications. Additionally, we will discuss determining antioxidant activity and total polyphenol content in wine.