Free amino acids (FAAs) are believed to play important roles in osmoregulation and buffer capacity in some aquatic animals, such as fishes. However, the potential roles of FAAs have not been systematically summarized and characterized until now. In the present study, the meta-analysis was conducted to investigate the relationships between FAAs and environmental salinities. Twenty published documents were included, accounting for 106 study cases. The effect sizes of total free amino acids (TFAAs), total essential amino acids (TEAAs), and total non-essential amino acids (TNEAAs) to salinity increase were calculated and determined by the restricted maximum likelihood (REML) method. It clearly showed that the elevated salinities significantly induced the contents of TFAAs, TEAAs, and TNEAAs at the ratio of 36%, 27%, and 29%, respectively. Faced to the salinity changes, the contents of FAAs in fishes under freshwater and seawater varied significantly, while the individuals under brackish water displayed relatively constant contents of FAAs. When salinity elevated, the contents of 17 amino acids in muscles significantly increased, suggesting the important roles of FAA metabolism in osmoregulation in fishes. The results also indicated that the effect sizes of TFAAs were positively related to the rates of salinity increases, and exhibited a significant quadratic linear relationship with temperatures. Additionally, the contents of FAAs also showed positive correlation with osmotic pressure, concentrations of plasma Na+, Cl-, and urea, implying their potential roles of FAAs in osmoregulation in fishes. These findings suggested that elevated salinities greatly induced the contents of FAAs in fishes, making a great contribution to maintaining the homeostasis of fishes in response to environmental salinity changes.