This study investigated metabolite changes in three pomelo cultivars during postharvest senescence using1H NMR-based metabolic profiling. Three pomelo cultivars, ‘Hongroumiyou’, ‘Bairoumiyou’ and ‘Huangroumiyou’, abbreviated as “R”, “W” and “Y” according to the color of their juice sacs, were stored at 25 °C for 90 days, and NMR was applied to determine the metabolite changes in juice sacs during storage. Fifteen metabolites were identified, including organic acids, sugars, amino acids, fatty acids, phenols and naringin. Partial least squares discriminant analysis (PLS-DA) was used to screen the significant metabolites according to the variable importance for the projection (VIP) scores in three pomelo cultivars during 90 days of storage. Additionally, eight metabolites, naringin, alanine, asparagine, choline, citric acid, malic acid, phosphocholine and β-D-glucose, were screened to be the crucial biomarkers with VIP > 1. The undesirable flavor of “bitter and sour” during the 60 days of storage was mainly attributed to the naringin, citric acid and sugars. According to the correlation analysis, the citric acid content determined by NMR showed a significantly positive relationship with that analyzed by HPLC. These findings suggested that NMR technology was accurate and efficient for metabolomic analysis of pomelo fruit, and the1H NMR-based metabolic profiling can be efficient during quality evaluation and useful for improving the fruit flavor quality during postharvest storage.