This article describes a new application of graphene oxide (GO) in CE based on the coating of fused silica capillary for chiral separation. The coated capillary was characterized by SEM, energy dispersive X-ray spectroscopy, and Raman spectra. The results indicated that the capillary was coated with GO. Chiral separations were carried out in the GO-coated capillary for the ephedrinepseudoephedrine (E-PE) isomers and -methylphenethylamine (-Me-PEA) isomers at the optimal buffer conditions without any chiral selector by CE. The precision and reproducibility of GO-coated capillary were investigated, and the RSDs of migration time (n = 6) for the E-PE isomers were 1.351.41%, and 0.973.50% for -Me-PEA isomers, respectively. The LOD for E-PE isomers and -Me-PEA isomers was 3 M and 18 M, respectively.