Several influences modulate biochemical responses to a weight-adjusted levothyroxine (L-T 4 ) replacement dose. We conducted a secondary analysis of the relationship of L-T 4 dose to TSH and free T 3 (FT 3 ), using a prospective observational study examining the interacting equilibria between thyroid parameters. We studied 353 patients on steady-state L-T 4 replacement for autoimmune thyroiditis or after surgery for malignant or benign thyroid disease. Peripheral deiodinase activity was calculated as a measure of T 4 -T 3 conversion efficiency. In euthyroid subjects, the median L-T 4 dose was 1.3 mg/kg per day (interquartile range (IQR) 0.94,1.60). The dose was independently associated with gender, age, aetiology and deiodinase activity (all P!0.001). Comparable FT 3 levels required higher L-T 4 doses in the carcinoma group (nZ143), even after adjusting for different TSH levels. Euthyroid athyreotic thyroid carcinoma patients (nZ50) received 1.57 mg/kg per day L-T 4 (IQR 1.40, 1.69), compared to 1.19 mg/kg per day (0.85,1.47) in autoimmune thyroiditis (P!0.01, nZ76) and 1.08 mg/kg per day (0.82, 1.44) in patients operated on for benign disease (P! 0.01, nZ80). Stratifying patients by deiodinase activity categories of !23, 23-29 and O29 nmol/s revealed an increasing FT 3 -FT 4 dissociation; the poorest converters showed the lowest FT 3 levels in spite of the highest dose and circulating FT 4 (P!0.001). An L-T 4 -related FT 3 -TSH disjoint was also apparent; some patients with fully suppressed TSH failed to raise FT 3 above the median level. These findings imply that thyroid hormone conversion efficiency is an important modulator of the biochemical response to L-T 4 ; FT 3 measurement may be an additional treatment target; and L-T 4 dose escalation may have limited success to raise FT 3 appropriately in some cases.