Interpretation of thyroid function tests during pregnancy needs trimester-related reference intervals from pregnant populations with minimal risk for thyroid dysfunction. While India has become iodine sufficient after two decades of salt iodisation, there is no normative data for thyroid function from healthy pregnant women of this country. To determine trimester-specific reference ranges for free triiodothyronine (FT(3)), free thyroxine (FT(4)) and thyrotropin (TSH) from healthy pregnant Indian women. Cross-sectional study in a reference population of pregnant women. Primary care level obstetric department in India. Women with uncomplicated pregnancy in any trimester. Five hundred and forty-one apparently healthy pregnant women with uncomplicated single intrauterine gestations reporting to the Armed Forces Clinic in any trimester were consecutively recruited. Clinical examination, thyroid ultrasound for echogenicity and nodularity and estimation of FT(3), FT(4), TSH and antithyroid antibodies (antithyroperoxidase [anti-TPO] and antithyroglobulin [anti-Tg]) using electrochemiluminescence technique were carried out. From this entire sample, a disease- and risk-free reference population was obtained by excluding those with any known factor that could affect thyroid function or those who were being treated for thyroid dysfunction. None. Of the 541 consecutive pregnant women in different trimesters enrolled for the study, 210 women were excluded. The composition of reference population comprising 331 women was 107 in first trimester, 137 in second trimester and 87 in third trimester. The 5th and 95th percentiles values were used to determine the reference ranges for FT(3), FT(4) and TSH. The trimester-wise values in the first, second and third trimesters were: FT(3) (1.92-5.86, 3.2-5.73 and 3.3-5.18 pM/l), FT(4) (12-19.45, 9.48-19.58 and 11.32-17.7 pM/l) and TSH (0.6-5.0, 0.44-5.78 and 0.74-5.7 iu/ml), respectively. Analysis of mean, median values for FT(3), FT(4) and TSH between each trimester showed no significant difference in FT(3) and TSH values (95% CI). However, FT(4) showed significant variation between trimesters with values decreasing with advancing gestational age (P value: first versus second = 0.015, first versus third = 0.003 and second versus third = not significant). Women with antibody positivity and hypoechogenicity of thyroid gland had significantly higher TSH values when compared with women with antibody negativity and normoechogenicity. Reference ranges of FT(3), FT(4) and TSH have been established for pregnant Indian women using 5th and 95th percentiles.