Renal failure is one among the slowly progressive diseases of kidney function characterized generally by low glomerular filtration (GRF). The replacement therapy of renal failure by hemodialysis involves the removal of excessive toxic fluids and toxic metabolic end products from the body. In this background, the present attempt was focused to evaluate and correlate the value of various biochemical markers in blood serum from the pre and post dialysis renal failure patients. Among many biochemical parameters in blood, serum creatinine and urea are emerging as a source of more sensitive markers for the detection of the renal failure. Thus, the analytical part of this study included the samples from 50 patients with renal failure before and after hemodialysis for the prevalence of creatinine and urea level. A colorimetrical method using fully automated analyser (Cobas C311) for the analysis of creatinine and urea concentrations in human serum was adopted. The mean values of these two components in post dialysis group were lower when compared to the pre dialysis group, and the concentration of urea was found to be lower than that of creatinine. The incidence of serum creatinine and serum urea was significantly high before hemodialysis and reduced significantly after hemodialysis. The study revealed a significant statistical relationship between the availability of creatinine and urea in blood serum of the pre and post dialysis samples from renal failure patients.