Perfluorobutanesulfonic acid (PFBS) was used to replace perfluorooctanesulfonic acid (PFOS) applications because it was less hazardous and had a shorter half-life than PFOS. The accumulation of PFBS in diverse environmental and humans has been steadily growing, and PFBS has caused ecological harm to aquatic species. Inevitably, environmental concerns about PFBS have grown, and the safety of PFBS is one of the most important current discussions, as the toxicity mechanism remains largely unknown. In this study, changes in protein expression of zebrafish larvae exposed to PFBS was investigated using Tandem mass tags (TMT)-labeled liquid chromatography mass spectrometry (LC-MS/MS) based quantitative proteomic analysis. A total of 1,959 protein were identified and 1,955 protein were quantified in zebrafish larvae exposed to PFBS. The volcano plot indicated differentially expressed protein (DEP) among these proteins. The number of DEPS at 7, 70, 700, 3500 and 7000 mM PFBS were determined to be 1, 1, 2, 3 and 14 respectively. As a result of cluster analysis, proteins in cluster 4 and 5 related to mitochondria function, structural proteins such as troponin and myosin and RNA functions related to ribosomes gradually decreased to 3,500 mM and then increased at the highest concentration of 7,000 mM. Furthermore, Cluster 8 shows that metabolic-related proteins such as cytochrome P450 and glutathione include in a concentration dependently increased pattern after sharp decreased at 7 mM. Among the parvalbumin proteins that were changed in the previous PFOS exposure study, parvalbumin increased 1.6-2.7 times according to PFBS exposure, suggesting that, like PFOS, it may be related to the neurotoxicity of PFBS. Overall, this research would help research better understand the mechanisms driving PFBS-mediated developmental neurotoxicity, as well as monitor and evaluate PFBS contamination in the environment.