ABSTRACT:
The simultaneous estimation of Levothyroxine (T4) and Liothyronine (T3) in combined pharmaceutical dosage formsis crucial for effective hypothyroidism management.Due totheir lowconcentrations,structural similarity, and high potency, method development poses analytical challenges. Reversed-phase high-performance liquid chromatography (RP-HPLC) stands out as a preferred technique owing to its high sensitivity, resolution, and reproducibility across a wide range of compound polarities.
This review outlines RP-HPLC principles and highlights method development strategies such as columns election, mobile phase optimization, detection wavelength, and sample preparation. It also discusses essential method validation parameters in line with ICH guidelines, including accuracy, precision, specificity, linearity, LOD, LOQ, robustness, and system suitability. Literature-based insights into validated methods for routine analysis, stability studies, and pharmacokinetic evaluations are presented.
Advancements such as gradient elution, stability-indicating approaches, and mass spectrometry integration are explored for their impact on method performance. The review concludes with clinical and regulatory perspectives, reinforcing RP-HPLC as a reliable tool for the quantitative analysis of thyroid hormones in pharmaceutical formulations.
1. Anusha,J.,
& Narasimhulu, G.(2024).Method Development and Validation
fortheSimultaneous Estimation of Levothyroxine and Liothyronine in Bulk and
Pharmaceutical Dosage Forms by RP-HPLC. Zenodo/ Research Gate.
2. Rapaka, R. S., Peffley, J. J., Doran, T. C.,
& Rowe, T. (1981). Dissolution and content uniformity of liothyronine
sodium and levothyroxinesodium tablets. Journal of Pharmaceutical Sciences,
70(5), 559–561.
3. Rai, A. K., & Yadav, S. (2024).
Simultaneous Estimation Method Development and Validation of Levothyroxine and
Liothyronine by HPLC Method. Asian Journal of Biological and Life Sciences,
13(1), 120– 125.
4. Abhay Kumar Rai et al. (2024). Simultaneous
Estimation Method Development and Validation of Levothyroxine and Liothyronine
by HPLC Method. Asian Journal of Biological and Life Sciences.
5. Rai, A. K., & Yadav, S. (2024).
Analytical Validation of Stability-Indicating Reverse Phase HPLC Method for
Simultaneous Estimation of Levothyroxine and Liothyronine. Asian Journal of
Biological and Life Sciences, 13(1), 126–132.
6. Dutt, R., Kaul, R., Bhat, P. A., &
Verma, P. (2020). Development and validation of UPLC-MS/MS method for rapid
simultaneous determination of levothyroxine and liothyronine in human serum.
Drug Delivery, 27(1), 426–434.
7. Collier, J.W., Nasr, M.M., &Mehvar, R.
(2011). A selective and sensitive gradient HPLC methodfor analysis of
levothyroxine sodium tablets. International Journal of Pharmaceutical Sciences
and Research, 2(9), 2315–2320.
8. Gika, H. G., Samanidou, V. F.,
&Papadoyannis, I. N. (2005). Development and validation of a HPLC method
for the determination of iodotyrosines and iodothyronines in biological
samples. Journal of Chromatography B, 819(1), 57–64.
9. Bunevicius, R., Kazanavicius, G.,
Zalinkevicius, R., & Prange Jr, A. J. (2002). Effects of thyroxine as
compared withthyroxineplustriiodothyroninein patientswith hypothyroidism.The
New England Journal ofMedicine, 340(6), 424–429.
10. Appelhof, B. C., Fliers, E., Wiersinga, W. M.,
et al. (2005). Combined therapy versus monotherapy inprimary hypothyroidism: a
double-blind randomized controlled trial. Journal of Clinical Endocrinology
& Metabolism, 90(5), 2666–2674.
11. Hennemann, G., Docter, R., Friesema, E. C. H.,
De Jong, M., & Krenning, E. P. (2004). Pharmacokinetics and patient
preferences of a new combined preparation of levothyroxine plus
sustained-release liothyronine. European Journal of Endocrinology, 150(1),
113–120.
12. Abhay Kumar Rai et al. (2024). Analytical
Validation of Stability-Indicating Reverse Phase HPLC Method for Simultaneous
Estimation of Levothyroxine and Liothyronine. Asian Journal of Biological and
Life Sciences.
13. Rohit Dutt et al. (2020). Development and
Validation of UPLC-MS/MS Method for Rapid Simultaneous Determination of
Levothyroxine and Liothyronine in Human Serum. Drug Delivery Journal.
14. Bhadru B, Rao TR, Jadav B. 2023. UV
Spectrophotometric Method development and validation for Metformin
Hydrochloride in bulk and its tablet formulation. Chemistry Research Journal 8
(6), 10-15.
15. Bhadru VNB, Rao V. 2023. Simultaneous
estimation of Levodopa and Benserazide by using reverse phase High Performance
Liquid Chromatography in API and marketed formulation, International Journal of
Recent Advances in Multidisciplinary Research 10: 9020-9024.
16. Boggula SKN, Bhadru B, Pathakala N, Peddapalli
H. 2023. Method development and validation of RP-HPLC method for the estimation
of Tolvaptan in bulk and its tablet dosage form. European Chemical Bulletin, 12
(10):5158-5171.
17. Patel S, Soni P, Omray LK. 2023. Development
and validation of RP-HPLC method for simultaneous estimation of Metformin
Hydrochloride and Gliclazide in bulk and tablet formulation. International
Journal of Drug Delivery Technology. 13(2):483-487.
18. Pravalika M, Lavanya J. 2023. Analytical
method development and validation of simultaneous estimation of Mifepristone
and misoprostol in bulk drug and pharmaceutical dosage form by RP-HPLC method.
International Journal of Research and Analytical Reviews 10(4): 674-690.
19. Pravalika P, Jephthah G, Reddy AR, Rao TR.
2023. A New RP-HPLC method for estimation of Vericiguat in bulk drug and
pharmaceutical dosage form. Journal of Advanced Scientific Research 14
(09):37-43.
20. Reddy AR, Sushma G., Rama Rao T., Kalyan
Varma V, Neelaveni K. 2023. Method development and validation of Tivozanib By
RP-HPLC in bulk and pharmaceutical dosage forms. International Journal of
Advanced Research, 11:566-573.