Analytical Method Development and Validation of RP-HPLC Method for Estimation of Levothyroxine in Bulk and Pharmaceutical Dosage Form
DOI:
https://doi.org/10.63682/jns.v14i32S.8146Keywords:
Levothyroxine sodium, RP-HPLC, Method Validation, ICH Q2(R1), Assay, LOD, LOQAbstract
Objective: To develop and validate a simple, accurate, and robust RP-HPLC method for the estimation of levothyroxine in bulk drug and pharmaceutical dosage forms, ensuring compliance with regulatory quality standards.
Method: A reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed using a C18 column (250 mm × 4.6 mm, 5 µm) with a mobile phase of phosphate buffer (pH 3.0) and methanol (55:45 v/v). The flow rate was set at 1.0 mL/min, detection wavelength at 225 nm, and injection volume was 10 µL. The method was validated as per ICH Q2(R1)1guidelines.
Result: The method exhibited excellent linearity in the range of 0.08–0.8 µg/mL with a correlation coefficient (r²) of 0.999. Accuracy was within 95–105%, precision (%RSD) was less than 2%, LOD was 0.03 µg/mL, and LOQ was 0.09 µg/mL. The method also demonstrated strong specificity and robustness.
Conclusion: The validated RP-HPLC method is simple, sensitive, precise, and suitable for the routine quality control and stability analysis of levothyroxine sodium in pharmaceutical formulations, meeting all regulatory validation criteria.
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ICH Q2(R1). Validation of Analytical Procedures: Text and Methodology. International Conference on Harmonisation, 2005.
Tang R, Wang J, Yang L, Ding X, Zhong Y, Pan J, Yang H, Mu L, Chen X, Chen Z: Subclinical Hypothyroidism and Depression: A Systematic Review and Meta-Analysis. Front Endocrinol (Lausanne). 2019 Jun 4;10:340. doi: 10.3389/fendo.2019.00340. eCollection 2019.
American Association of Clinical Endocrinologists (AACE): Clinical Practice Guidelines for Hypothyroidism in Adults.
Jonklaas J, Bianco AC, Bauer AJ, Burman KD, Cappola AR, Celi FS, Cooper DS, Kim BW, Peeters RP, Rosenthal MS, Sawka AM: Guidelines for the treatment of hypothyroidism: prepared by the american thyroid association task force on thyroid hormone replacement. Thyroid. 2014 Dec;24(12):1670-751. doi: 10.1089/thy.2014.0028.
Kaplan MM, Breitbart R: Conversion of thyroxine to triiodothyronine in the anterior pituitary gland and the influence of this process on thyroid status. Horm Metab Res Suppl. 1984;14:79-85.
Osmak-Tizon L, Poussier M, Cottin Y, Rochette L: Non-genomic actions of thyroid hormones: Molecular aspects. Arch Cardiovasc Dis. 2014 Apr;107(4):207-11. doi: 10.1016/j.acvd.2014.02.001. Epub 2014 Mar 26.
Klein I, Ojamaa K: Thyroid hormone and the cardiovascular system. N Engl J Med. 2001 Feb 15;344(7):501-9. doi: 10.1056/NEJM200102153440707.
Indian Pharmacopoeia, 2022.
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