Introduction: Spherical particles with nonpolar neutral lipids at the center and more polar amphipathic lipids at their surface are called Lipoproteins. HDL aids in cholesterol homeostasis by removing it from cells by several different mechanisms. The results of HDL Cholesterol (HDL-C) should be accurate and reliable, especially for the patients with borderline values, where the variation resulting from the methodology used may interfere with the interpretation of clinical scenario by the doctors. Aim: To compare the results of HDL values obtained by using three different technical methods. Materials and Methods: The present study was conducted using 30 serum samples from the Biochemistry laboratory of KLE Society’s Dr. Prabhakar Kore Charitable Hospital and Medical Research Centre, Belagavi, Karnataka, India for HDL values, each for the three different technical methods (fullyautomated, semi-automated and manual method). The data obtained was analysed statistically by computing descriptive statistics the mean, standard deviation and correlation coefficient. Results: Results of serum samples analysed for HDL-C values ranged from 31-79 mg/dl (mean 43.1) in fully-automated analyser, 28.06 to 75.18 (mean 39.6) in semi-automated and 27.06 to 62.7mg/dl (mean 39.7) using manual method. There was significant difference in the mean values of HDL-C values obtained using the three different methods and positive correlation was established on comparing semi-automated and manual method (Semi-automated v/s manual method, r=0.827) and found to be statistically significant. Conclusion: As HDL estimation is routinely done in many of the clinical laboratories, a sound knowledge of the HDL values obtained using different techniques can help the clinicians while ordering for the test and also for diagnosing & monitoring of the cases with various lipid abnormalities.