Development and Validation of UPLC-MS/MS Method for Rapid Simultaneous Determination of Levothyroxine and Liothyronine in Human Serum

  • Rohit Dutt, Prof Department of Pharmacy, School of Medical and Allied Sciences, G D Goenka University, Gurugram-122103, India
  • Kailash Chander Malik, Mr Department of Pharmacy, School of Medical and Allied Sciences, G D Goenka University, Gurugram-122103, India
  • Manoj Karwa, Dr Auriga Research Private Limited, Plot no. 136, Sector-5, IMT Manesar, Gurugram, India
  • Gaurav Kumar JAIN Department of Pharmaceutics, School of Pharmaceutical Sciences, Delhi Pharmaceutical Science and Research University, New Delhi, India
DOI https://doi.org/10.22270/jddt.v10i3-s.4189

Abstract

A simple ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method was developed and fully validated to simultaneously determine levothyroxine (LT4) and liothyronine (LT3) in human serum. Sample preparation was done through protein precipitation with acetonitrile. HyPURITY C18 column was selected to achieve rapid separation for LT4 and LT3 within 4 min. Electrospray ionization (ESI) under multiple reaction monitoring (MRM) was used to monitor the ion transitions for LT4 (m/z 777.54→731.52), LT3 (m/z 651.64→ 605.65) and internal standard LT4-D3 (m/z 780.53 →734.19), operating in the positive ion mode. The method was proved to be accurate (82.35% to 113.56%) and precise (0.73% to 8.28%) over concentration range of 50.37 ng/ml – 300.13 ng/ml for LT4 and 0.5 ng/ml – 50.37 ng/ml for LT3. The validated method could be applied for pharmacokinetic study or bioequivalence testing of combination products of LT4 and LT3.


Keywords: Levothyroxine; Liothyronine; Ultra Performance Liquid Chromatographic; Mass Spectrometry; Human Serum

Keywords: Levothyroxine, Liothyronine, Ultra Performance Liquid Chromatographic, Mass Spectrometry, Human Serum

Downloads

Download data is not yet available.

Author Biographies

Rohit Dutt, Prof, Department of Pharmacy, School of Medical and Allied Sciences, G D Goenka University, Gurugram-122103, India

Department of Pharmacy, School of Medical and Allied Sciences, G D Goenka University, Gurugram-122103, India

Kailash Chander Malik, Mr, Department of Pharmacy, School of Medical and Allied Sciences, G D Goenka University, Gurugram-122103, India

Department of Pharmacy, School of Medical and Allied Sciences, G D Goenka University, Gurugram-122103, India

Manoj Karwa, Dr, Auriga Research Private Limited, Plot no. 136, Sector-5, IMT Manesar, Gurugram, India

Auriga Research Private Limited, Plot no. 136, Sector-5, IMT Manesar, Gurugram, India

Gaurav Kumar JAIN, Department of Pharmaceutics, School of Pharmaceutical Sciences, Delhi Pharmaceutical Science and Research University, New Delhi, India

Department of Pharmaceutics, School of Pharmaceutical Sciences, Delhi Pharmaceutical Science and Research University, New Delhi, India

References

1. Taylor PN, Albrecht D, Scholz A, Gutierrez-Buey G, Lazarus JH, Dayan CM, et al. Global epidemiology of hyperthyroidism and hypothyroidism. Nat Rev Endocrinol. 2018; 14(5):301–16.
2. Jacqueline Jonklaas, Bianco AC, Bauer AJ, Kenneth D. Burman, Cappola AR, Celi FS, et al. GUIDELINES FOR THE TREATMENT OF HYPOTHYROIDISM Prepared by the American Thyroid Association Task Force on Thyroid Hormone Replacement Division of Endocrinology , Georgetown University, Washington, DC ; 5 Division of Endocrinology , Rush University Medic. Thyroid. 2014; 24(12):1670–751.
3. Peterson SJ, McAninch EA, Bianco AC. Is a normal TSH synonymous with “euthyroidism” in levothyroxine monotherapy? J Clin Endocrinol Metab. 2016;101(12):4964–73.
4. Escobar-Morreale HFF, Botella-Carretero JI, De Escobar GM. Treatment of hypothyroidism with levothyroxine or a combination of levothyroxine plus L-triiodothyronine. Best Pract Res Clin Endocrinol Metab. 2015; 29(1):57–75.
5. Clyde PW, Shakir KMM. in Primary Hypothyroidism. 2017; 290(22).
6. Biondi B, Wartofsky L. Combination treatment with T4 and T3: Toward personalized replacement therapy in hypothyroidism? J Clin Endocrinol Metab. 2012; 97(7):2256–71.
7. Escobar-Morreale HF, Obregón MJ, Hernández A, Escobar Del Rey F, Morreale De Escobar G. Regulation of iodothyronine deiodinase activity as studied in thyroidectomized rats infused with thyroxine or triiodothyronine. Endocrinology. 1997; 138(6):2559–68.
8. Wiersinga WM. Paradigm shifts in thyroid hormone replacement therapies for hypothyroidism. Nat Rev Endocrinol [Internet]. 2014; 10(3):164–74. Available from: http://dx.doi.org/10.1038/nrendo.2013.258
9. Chopra IJ, Santini F, Hurd RE, Chua Teco GN. A radioimmunoassay for measurement of thyroxine sulfate. J Clin Endocrinol Metab. 1993; 76(1):145–50.
10. Shah RB, Bryant A, Collier J, Habib MJ, Khan MA. Stability indicating validated HPLC method for quantification of levothyroxine with eight degradation peaks in the presence of excipients. Int J Pharm. 2008; 360(1–2):77–82.
11. Kazemifard AG, Moore DE, Aghazadeh A. Identification and quantitation of sodium-thyroxine and its degradation products by LC using electrochemical and MS detection. J Pharm Biomed Anal. 2001; 25(5–6):697–711.
12. Bajaj S, Singla D, Sakhuja N. Stability testing of pharmaceutical products. J Appl Pharm Sci. 2012; 2(3).
Statistics
127 Views | 105 Downloads
How to Cite
1.
Dutt R, Malik K, Karwa M, JAIN G. Development and Validation of UPLC-MS/MS Method for Rapid Simultaneous Determination of Levothyroxine and Liothyronine in Human Serum. JDDT [Internet]. 15Jun.2020 [cited 17Jan.2021];10(3-s):176-81. Available from: http://jddtonline.info/index.php/jddt/article/view/4189
Issue
Vol 10 No 3-s (2020): Volume 10, Issue 3-s, May-June 2020 (Supplement Issue)
Section
Research
Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Authors who publish with this journal agree to the following terms:

  1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
  2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
  3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).