In this study, graphene oxide (GO) was synthesized by the improved Hummers’ method. The degree of oxidation from graphite (Gi) to GO was determined through interlayer spacing calculated from X–ray diffraction. Besides, the effect of KMnO4: Gi ratios (X1), H2SO4 volume (X2), oxidation temperature (X3), oxidation time of stage 1 (X4), and oxidation time of stage 2 (X5) was screened by the Plackett–Burman model. The simultaneous impact of three factors that influenced the degree of oxidation (X1, X2, and X3) was studied by the Box–Behnken experimental model of response surface methodology to achieve suitable conditions for the GO synthesis process. The characterization of GO product was investigated via the modern analytical methods: X-ray diffraction, Raman spectroscopy, Fourier transform infrared spectroscopy, UV–Vis spectroscopy, field emission scanning electron microscopy, transmission electron microscopy, and atomic force microscopy. Inaddition, the study was also carried out on a pilot scale for orientation in industrial application with the yield of 14 g/batch.