Levothyroxine is a synthetic of thyroxine (T4), a key thyroid hormone, and is widely
used in the treatment of hypothyroidism and other endocrine disorders. Due to its narrow
therapeutic window and the necessity for accurate dosing, the application of reliable and
validated analytical techniques is critical to ensure its safety, quality, and therapeutic
effectiveness. This provides a comparative evaluation a 10analytical methods
employed for the quantification of levothyroxine in both pharmaceutical formulations and
biological samples. The reviewed techniques include Ultraviolet (UV)
Spectrophotometry, derivative UV analysis, High-performance liquid
chromatography (HPLC), Ultra-high performance liquid
chromatography (UHPLC), High-performance thin-layer chromatography (HPTLC),
Capillary electrophoresis and Tandem Mass Spectrometric methods such as LC-MS/MS
and UPLC-MS/MS. These approaches utilize various stationary phases, including C18,
BEH C18, and silica gel, in combination with mobile phases comprising methanol,
acetonitrile, formic acid, and buffers of varying pH. The detection wavelengths range from 200 to 254 nm, with flow rates between 0.2 and 1.5 mL/min. Notably, LC-MS/MS
and UPLC-MS/MS demonstrate superior sensitivity and specificity, making them highly
effective for trace-level detection in complex matrices. This comprehensive summary
serves as a practical guide for selecting suitable analytical techniques based on the
intended application in pharmaceutical analysis or clinical evaluation.
Keywords: Levothyroxine, Analytical Techniques, UV Spectrophotometry, RP-
HPLC, LC-MS/MS, Method Validation, Thyroid Hormone Analysis
Publication date: 01/08/2026
https://www.ijbpas.com/pdf/2026/August/MS_IJBPAS_2026_10333.pdf
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https://doi.org/10.31032/IJBPAS/2026/15.8.10333
