Purpose: To evaluate the role of "penumbra sign", diffusion-weighted imaging (DWI), and the apparent diffusion coefficient (ADC) value in differentiating tubo-ovarian abscess (TOA) from ovarian malignancy. Material and methods: Thirty-six patients with 50 adnexal masses (tubo-ovarian abscess, n = 24; ovarian malignancy, n = 26), who underwent magnetic resonance imaging (MRI) with DWI, were retrospectively evaluated. "Penumbra sign" (hyperintense rim on T1W images), diffusion restriction, and mean apparent diffusion coefficient (ADC) values from cystic (c-ADC) and solid (s-ADC) components were evaluated for all the masses. Results: "Penumbra sign" on T1W images was significantly more common in the TOA group (n = 21, 87.5%) than in the ovarian malignancy group (n = 2, 7.7%) (p < 0.001). Similarly, diffusion restriction in the cystic component was more frequent in the TOA group (n = 24, 100% vs. n = 2, 10.5%; p < 0.001). In contrast, diffusion restriction in the solid component was more common in the ovarian malignancy group (n = 5, 20.8% vs. n = 26, 100%; p < 0.001). The mean c-ADC value was significantly lower in TOAs (p < 0.001). A c-ADC value of 1.31 × 10-3 mm2/s may be an optimal cut-off in distinguishing TOAs from ovarian malignancies. Conversely, the mean s-ADC value was significantly lower in the ovarian malignancy group (p < 0.001). An s-ADC value of 0.869 × 10-3 mm2/s may be an optimal cut-off in differentiating ovarian malignancies from TOAs (p < 0.001). ROC curve analysis showed that c-ADC values had a higher diagnostic accuracy than s-ADC values. Conclusions: "Penumbra sign" on T1W images, diffusion characteristics, and ADC values provide important clues in addition to conventional MR imaging features in differentiating TOA from ovarian malignancy.