Carbohydrates, including monosaccharides, oligosaccharides and polysaccharides, are widely distributed in food resources. Matrix-assisted laser desorption/ionization (MALDI), a soft ionization technique, is extensively applied in carbohydrates analysis. MALDI mass spectrometers are capable to generate spectra rapidly and record mass profiles or fingerprints from foods, usually with simple sample preparation or pretreatment. Therefore, this review described recent research progress on MALDI mass spectrometry (MS) detection in food carbohydrates, especially for the improvement of ionization efficiency and their application in structure characterization of polysaccharides from food and edible herbs. Furthermore, the quantitative study and mass spectrometry imaging were also briefly summarized.