The Bio-Rad Variant Hemoglobin Testing System is an automated HPLC analyzer marketed with a Beta-thalassemia Short Program to quantify hemoglobins (Hbs) F and A2 and assist in detecting Hbs A, S, C, D, and E. We evaluated this system to replace several traditional methods for Hb in our hospital laboratory. Analytical performance relevant to quantifying Hbs A, S, C, and F was assessed with blood samples obtained from our local patient population. Studies of precision (CVs < 3%) and analytical limits (% of total Hb) of Hbs A (2-86%), F (1-89%), S (5-90%), and C (3-92%) demonstrated results comparable with or exceeding those of traditional methods. Results for patients' samples (n) for Hbs A (107), F (157), S (128), and C (27) correlated well (r > 0.93) with results by traditional methods. The satisfactory performance and efficiency led us to implement this system for routine quantification of clinically significant Hbs.