The measurement of glycohemoglobin is the best measure of mean glucose within a 3-4 month range. As it is used for patient education, counseling, feedback control, and ultimately for patient motivation, its measurement should be optimally accurate and precise. Duplicate hemoglobin A1c readings were used to determine physiological (changes over time between measurements) and analytic variation of two widely used laboratory assays: Bio-Rad Variant II's high-performance liquid chromatography (HPLC) system and Roche's immunoassay. The average variation of grouped duplicates was calculated and graphed against corresponding time intervals. Regression to the y-intercept (0 day separation between readings) was used to determine the analytic variation. Analytic coefficients of variation (CVs) for the HPLC and immunoassay were determined as 2.6% and 5.1%, respectively. The CV of the immunoassay method exceeds physiologically established limits of 2-3% and those of the National Glycohemoglobin Standardization Program (3-4%). The Bio-Rad HPLC system produces a CV within these limits.