The aim of this study was to assess biochemical markers of rabbit amniotic fluid during fetal growth stage of successful pregnancy. Experiments were executed on 15 pregnant rabbits at three different time point of successful pregnancy i.e. day 18, day 25 and day 28 with commercially accessible kits for biochemical markers that includes glucose, total protein, albumin, creatinine, triglycerides, blood urea nitrogen (BUN), alanine transaminase (ALT) and aspartate transaminase (AST). The current study demonstrated the ascertained value in amniotic fluid for glucose, total protein and albumin at day 18, 25 and 28 was 42.9±0.80, 37.4±0.90, 36.5±0.88mg/dl; 17.9±0.30, 16.1±0.22, 13.9±0.34g/l; 13.9±1.6,11.5±0.75, 8.8±0.15g/I whereas that of creatinine, triglyceride, BUN, ALT and AST at the corresponding days of pregnancy were 1.5±0.1, 1.6±0.2, 1.7±0.15mg/dl; 6.06±0.15, 4.6±1.2, 3.2±0.8mg/dl, 13.1±0.02, 14.2±0.3, 18.5±0.3mg/dl; 36.5±0.6, 32.8±0.5,17.3±1.2 IU/l; 7.9±1.3, 8.7±0.4, 14.8±1.9 IU/l. During current study, glucose, total protein, Albumin, triglyceride and ALT was significantly down regulated whereas creatinine, BUN, and AST was up regulated in the amniotic fluid at the end of fetal growth of successful pregnancy in rabbits.