Background: The significance of investigation for diagnosing and managing thyroid dysfunction in pregnant females has been extensively documented in the medical literature. This study aimed to determine trimester-specific reference ranges for thyroid-stimulating hormones (TSH), Free T3 (FT3), and free T4 (FT4) in apparently healthy pregnant women attending tertiary care hospitals in Lahore. Methods: This cross-sectional study was conducted at two tertiary care Hospitals in Lahore, Pakistan. In this multi-centric study, initially, 500 pregnant females were enrolled from September 2019 to December 2019 who fulfilled the inclusion criteria. For measurement of serum freeT3, free T4, TSH, anti-TPO, and Thyroglobulin antibodies, 5 ml of the blood sample was drawn, under aseptic conditions, from each subject using Maglumi 800 chemiluminescence immunoassay (CLIA) system. Results: Out of 500 subjects, 23 subjects with positive anti-TPO, 19 subjects with anti-TG antibodies, and 12 subjects due to less volume of serum yielded from whole blood (serum less than 3 ml) were excluded from the analysis. Ten samples were hemolyzed and not included in the analysis. A total of 436 samples were examined for analysis. Of the remaining 436 subjects, 133 (30.5%) were from 1st Trimester, 153 (35.1%) from 2nd Trimester, and 150 (34.4%) from 3rd Trimester. As the data were non-normal, the 2.5th, 50th and 97.5th percentiles were calculated to express each group's results. Trimester specific range of TSH 0.168-4.294, 0.258-4.584 and 0.341-4.625, FT3 1.857-4.408, 1.958-4.621 and 2.025-4.821 pg/mL and FT4 8.815-18.006, 8.306-17.341 and 7.402-17.292 pg/mL. Conclusion: In this study, we established a trimester-specific reference range for our local population's thyroid function test. The results of this study have complemented the results of previous studies.