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

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

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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

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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 2Jul.2020];10(3-s):176-81. Available from: http://jddtonline.info/index.php/jddt/article/view/4189