INTRODUCTION: Biological variation (BV) data are necessary for interpretation of test results and assessment of analytical performance. We aimed to determine the BV estimates for thyroid stimulating hormone (TSH), free triiodothyronine (fT3) and free thyroxine(fT4) in healthy subjects in Turkey and compare them with the literature findings. MATERIALS AND METHODS: A total of 21 Turkish healthy volunteers (12 males and 9 females) were included in the study. Blood samples were collected once a week for five weeks, and the analysis was performed using the chemiluminescent immunoassay method on an Advia Centaur XP (Siemens Diagnostic, Tarrytown, USA). Analytical variation (CV RESULTS: For TSH, fT3 and fT4, CV CONCLUSIONS: Our study provides updated BV data for thyroid function tests (TFTs) in healthy subjects in Turkey. As TFTs have shown a high degree of individuality, RCV should be preferred rather than population-based reference ranges in the assessment of serum concentrations. Our BV estimates were compatible with European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) BV meta-analysis data obtained using different immunoassay methods in different populations.