Aim of the study was to find out the role of biochemical markers in diagnosis of alcoholic liver disease and non-alcoholic liver disease and compared these values with healthy subjects of Rajasthan. Total 300 subjects were selected in which 200 were selected as patients group (alcoholic liver disease (ALD) and non-alcoholic liver disease (NALD) group) and 100 were normal healthy subjects. Total 100 alcoholic liver disease patients in which 91 patients were male and 09 patients were female. In non-alcoholic liver disease group 55 males and 45 females are enrolled. Patients were distributed age wise. Serum cholinesterase and serum 5-nucleotidase were estimated by commercially available kit on spectrophotometer. Serum cholinesterase level is 2822.99± 716.51 in ALD patients. In NALD patients, serum cholinesterase level was 4299.95±1014.76 and in control group mean serum cholinesterase level was 6303.99±977.59. Pvalue was statically highly significant. (P≤ 0.0001), as well as serum 5-nucleotidase level was 25.89± 5.77 and 17.70± 4.43 in ALD and NALD respectively. In control group mean level of serum 5-nucleotidase was 9.851± 3.24. (P≤ 0.0001). We conclude that serum cholinesterase level was decreased in both disease group, whereas serum 5-nucleotidase level is increased in both disease groups. Both parameters are liver specific and specific test for alcohol induced or alcoholic liver disease as well as in non-alcoholic liver disease. Both parameters can be used as a marker of liver disease. it was concluded that serum cholinesterase and 5'-nucleotidase are better diagnostic marker then the conventional liver function test that are raised in other liver disease also.