This study is to examine the effect of graphene oxide (GO) and GO-based nanocomposites (GO_EDTA, GO_CHIT) on the smear layer by scanning electron microscopy (SEM). Sixty human single-rooted anterior teeth were used. Samples were randomly categorised into six groups. Distilled water in group 1, 17% EDTA in group 2, chitosan (CHIT) solution in group 3, GO solution in group 4, GO_EDTA solution in group 5 and GO_CHIT in group 6 were used as irrigation solutions. Photomicrographs were acquired from the coronal, middle and apical parts of the samples by SEM. Statistical analyses were performed using Kruskal-Wallis, Mann–Whitney U and Wilcoxon tests. There was a statistically significant difference among the groups (p < 0.003). Adding GO to CHIT and EDTA did not affect removing the smear layer in all segments (p < 0.003). Incorporating GO into EDTA and CHIT solutions did not change their interaction with the smear layer.