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Fetal fluid contents have functions in protecting fetuses and are essential for fetal development and maturation. However, little is known about the exact physiological functions of fetal fluids in fetal development, as well as the changing composition throughout the gestational period in cats. In this study, the biochemical composition of amniotic (AMN) and allantoic (ALL) fluids was investigated, as well as in the maternal serum of pregnant queens. Eighteen queens were included in this study and assigned to six different groups, D₂₀, D₂₅, D₃₀, D₄₀, D₄₅ and D₆₀, according to the gestational stage of fetal development. A total of 44 amniotic and 37 allantoic samples were collected. Fetal fluids contained lesser concentrations of alanine aminotransferase, albumin, cholesterol, triglycerides, creatine kinase, amylase, total protein and globulin than maternal serum. Other variables, such as aspartate aminotransferase, gamma-glutamyl transferase, bilirubin and alkaline phosphatase, were in different concentrations at specific stages of gestation when compared to maternal serum. There were no differences between fetal fluids and maternal serum for lactate dehydrogenase, urea, lipase or glucose concentrations. There were greater concentrations of creatinine in amniotic fluid than in allantoic fluid or maternal serum. Based on the results of this study, fetal fluids do not accumulate as a result of the simple filtration of maternal blood, but rather, the fetus produces many of these components as a consequence of organ development and maturation.