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Carbon nanowalls (CNWs), vertically standing highly crystallizing graphene sheets, were used in the application of a surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-TOF-MS). The CNW substrates solved the issues on interferences of matrix molecules and alkali metal addition ions in low-weight molecule detection. Before SALDI sample preparations, the hydrophobic CNW was treated by atmospheric pressure plasma for exposing hydrophilicity to the CNWs’ surface. Detection of water soluble amino acids, arginine, was demonstrated.