OPTIMIZATION AND AUTHENTICATION OF RP-HPLC PROCEDURE FOR DETERMINING SYNTHETIC THYROID HORMONE REPLACEMENT
Keywords:
Levothyroxine, Liothyronine, RP-HPLC, Mixed-Method Approach, Stability-Indicating Method, Thyroid Hormone AnalysisAbstract
This study presents the development, optimization, and validation of a robust reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of levothyroxine and liothyronine in pharmaceutical formulations. Levothyroxine (T4) and liothyronine (T3) are synthetic thyroid hormones crucial for managing hypothyroidism and other endocrine disorders. Given the sensitivity and low concentration of these hormones in formulations, precise analytical techniques are imperative. The optimized RP-HPLC method employed a C18 column with a mobile phase of acetonitrile and water (25:75 v/v), adjusted to pH 6.2, with UV detection at 300 nm. The method demonstrated excellent linearity (10–100 µg/mL), precision, accuracy, specificity, and robustness in accordance with ICH guidelines. Forced degradation studies under acidic, basic, oxidative, thermal, and photolytic conditions confirmed the method’s stability-indicating nature. Complementary UV spectrophotometric methods, including absorption correction and first-order derivative techniques, were also validated and applied. The results indicate high assay accuracy (within 99–101%) and minimal interference from excipients or degradation products. This validated RP-HPLC method ensures reliable quality control of thyroid hormone formulations, enhancing the safety and efficacy of endocrine pharmacotherapy.
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