Background: Diabetes mellitus (DM) refers to a group of common metabolic disorders that share the phenotype of hyperglycemia. Depending on the etiology of the DM, factors contributing to hyperglycemia include reduced insulin secretion, decreased glucose utilization, and increased glucose production [1]. Aims and Objective: To evaluate the performance of Enzymatic and HPLC assay systems for determination of HbA1c. Material and Methods: This study was conducted from November, 2016 to December, 2017. The subjects under the study were enrolled from outpatient department of Medicine after obtaining written informed consent in bilingual languages from all the 100 subjects after describing all pros and cons. Fasting Plasma glucose and HbA1c were estimated in Department of Biochemistry, SRMS IMS. Result and Discussion: Maximum percentages of patients were in age 51 to 60 years (32%) while the least percentage of patients was from the age group 21 to 30 years (2%). Present study comprising 38% females and 62% males. HbA1c distribution in subjects by Enzymatic method in Mean ± SD (8.598 ± 2.29) with minimum of 6.2% and maximum 20.4%. The FPG distribution in Subjects 154.37 ± 40.13 (Mean ± SD) with minimum FPG of 96 mg/dl and maximum 289 mg/dl. Conclusion: HbA1c has come to play an integral role in the management of diabetes, one of the world’s most prevalent non-communicable diseases. HbA1c defines an end point as the fuel of diabetic therapy and provides a powerful stimulus to the patients to improve their compliance. HbA1c remains the gold standard in the assessment of glycemic control with availability of standardize methods.