Introduction and objective: There are methods of HbA1c estimation that depend on different physical, chemical, immunological features of glycated hemoglobin. Numerous analytical techniques have been developed for assessing HbA1c in diabetes mellitus (DM), including immunoturbidimetry, boronate affinity chromatography, enzymatic assays, and high-performance liquid chromatography (HPLC) immunoassay. Notably, different estimation methods may yield disparate results. This study aims to conduct and compare two analytical techniques—specifically, the enzymatic method and HPLC method—to observe and analyze any variations in results within the same set of samples. Materials and methods: This is an observational cross-sectional study involving 100 EDTA samples. The study focused on the analysis of HbA1c using two distinct methods: Atellica CH 930 enzymatic hemoglobin A1c (HbA1c_E) from Siemens Healthineers and cation exchange HPLC from Bio-Rad Laboratories. Results: A strong robust correlation was observed between the two methods, as evidenced by a Pearson's Coefficient of 0.983. The Bland–Altman plot demonstrated a high level of agreement between the two techniques, with 95% of values falling within ±SD (standard deviation), indicating a strong concordance. Conclusion: This study establishes that both methods, Atellica CH 930 enzymatic hemoglobin A1c (HbA1c_E) and cation exchange HPLC, produced comparable results for HbA1c. Therefore, both analytical techniques are deemed suitable for the effective management of diabetes.