To evaluate the kernel quality characteristics of the local sesame resources in Guizhou, eight quality-related traits of 73 Guizhou sesame accessions were analyzed. The oil content of sesame germplasms was ranged from 41.45% to 52.12%, with an average of 49.69%. The mean value of unsaturated fatty acids oleic acid and linoleic acid was 35.65% and 50.66%, respectively, while that of saturated fatty acids including palmitic acid and stearic acid was 8.40% and 4.79%, respectively. Furthermore, mean of three components sesamin, sesamolin and lignin in the tested sesame accessions were 5.03, 2.63 and 4.79 mg/g, respectively. The variation coefficient of these eight quality traits ranged from 3.69% to 32.62%, while the highest and lowest values were observed in sesamin and oil contents, respectively. Notably, the variation coefficient of sesamin content, sesamolin content and stearic acid content was more than 10%, which indicated that these three traits were significantly different among sesame samples. The correlation analysis showed that oil content was significantly positively correlated with oleic acid content and sesamin content. However, the oil content was significantly negatively correlated with linoleic acid content. Moreover, oleic acid content was positively correlated with the sesamin content, but negatively correlated with linoleic acid content. The linoleic acid content was negatively correlated with the sesamin content. By Principal component analysis (PCA), three principal components including oleic acid factor, oil content factor and sesamin factor explained 96.59% of the phenotypic variation. The cluster analysis roughly revealed six groups (Euclidean distance D = 9.75), which include 2, 12, 5, 16 and 31 accessions shared in Group Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ and Ⅵ, respectively. The accessions resided in group Ⅵ contained the highest oleic acid and oil content as well as moderate sesamin content. Taken together, this study unlocked the quality-related characteristics of Guizhou sesame accessions, which could provide a theoretical basis and good references for utilization and innovation of sesame germplasms.