AbstractPhotoluminescent carbon dots (CDs) fractions have been isolated from a gel‐like material (GM), which was synthesized using a convenient one‐step solvothermal route. In terms of purification, size exclusion chromatography (SEC) and dialysis were compared with acetone wash, which revealed the advantage of acetone wash. The pre‐purified GM with acetone wash (A‐GM) was further isolated by the reversed‐phase preparative thin layer chromatography (TLC) with acetonitrile−water mixture (7 : 3; va/vw) as the developing solvent. As a result, there were four photoluminescent bands on the TLC plate, which indicated the presence of four photoluminescent fractions. Detailed characterization measurements such as UV/Vis absorption, fluorescence emission, attenuated total reflection Fourier‐transform infrared spectroscopy, X‐ray photoelectron spectroscopy, zeta potential, dynamic light scattering, atomic force microscopy, and TEM measurements were performed on all fractions to analyze their heterogeneous optical, structural, electrical, and morphological properties. Considering the comprehensive analysis, all isolated fractions were CDs. In addition, excitation wavelength‐independent CDs were obtained with a mean size of 2.5 nm and high quantum yield (55 %). Furthermore, the study demonstrates that the excitation wavelength‐dependent photoluminescence of GM could result from the mixture of different surface states of each CD fraction rather than multiple surface states of uniform CDs nanoparticles.