4-Hydroxy-2,5-dimethyl-3(2)-furanone (furaneol) is present in food. It has a caramel-like flavor, which affects the quality of food, and is formed via multiple pathways. Msalais is a traditional wine fermented from boiled local grape juice in Xinjiang (China). It has a strong caramel odor, which suggests high furaneol content. Furaneol formation during Msalais-making had not been investigated to date. Here, high-performance liquid chromatography and different fermentation models of Msalais-making were used to investigate the furaneol content and formation during Msalais-making. The furaneol content of Msalais is high, between 27.59 ± 0.493 mg/L and 117.6 ± 0.235 mg/L. It is formed throughout the entire Msalais-making process. The formation pathways include the Maillard reaction and chemical hydrolysis of bound furaneol during grape juice concentration; enzymatic release and/or chemical acidic hydrolysis of furaneol glucosides, and biosynthesis from Maillard products and d-fructose-1,6-diphosphate during fermentation; chemical transformation of Maillard products at room temperature (16-25 °C) and hydrolysis of furaneol glucosides during storage. Importantly, furaneol is formed by an efficient biotransformation of Maillard products. These findings suggest that furaneol content can be used as an important indicator of wine quality, and could be controlled by controlling the grape quality, grape juice concentration, fermentation, and wine storage.