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Development of Ultra High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Enantiomer Resolution of Thyroxine on a Chiral Crown Ether Derived Chiral Stationary Phase

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Abstract

Chiral discrimination of thyroxine (T4) enantiomers was performed using ultra high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) on a chiral crown ether-derived ChiroSil SCA (-) column. The different composition of mobile phases and the effect of column oven temperatures were investigated and the optimum chromatographic separation with respect to resolution and analysis time was achieved using a mixture of 60% methanol/water (v/v) with 0.1% formic acid at 40 °C having a flow rate of 1.4 mL min−1. The thermodynamic data from van’t Hoff plots of temperature experiments revealed that the enantioseparation was enthalpically favored process. The method was validated in the concentration range of 0.5–100 μg mL−1 for both enantiomers and proved to be rapid, precise, sensitive, and selective method for the enantiodiscrimination of T4 under the optimized conditions. The calibration curves of both D- and L-T4 showed an excellent linearity with coefficient of determination (R2) > 0.9997. The developed chiral method was successfully applied for a quantitative assay to check the enantiomeric purities of the six pharmaceutical formulations of levothyroxine sodium tablets and the enantiomeric impurities identified were in the range of 0.11–0.29%. This method could be applied for the determination of enantiomeric purity on pharmaceuticals and also for the monitoring of thyroid hormone levels.

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Acknowledgements

This research was supported by Health Fellowship Foundation, 2021, South Korea.

Funding

Health Fellowship Foundation, 2021

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Author notes

  1. Jisun Lee and Suraj Adhikari contributed equally to this work.

Authors and Affiliations

  1. College of Pharmacy, Duksung Women’s University, Seoul, 01369, South Korea

    Jisun Lee & Hye-Ran Yoon

  2. College of Pharmacy, Chosun University, Gwangju, 61452, South Korea

    Suraj Adhikari & Wonjae Lee

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  1. Jisun Lee
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  2. Suraj Adhikari
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  3. Wonjae Lee
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  4. Hye-Ran Yoon
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Contributions

Experiment, data collection and analysis were performed by J. Lee. Interpretation of the data and writing of the original manuscript was done by S. Adhikari. Conceptualization, experimental design, editing and supervision were contributed by W. Lee. Analysis tools, financial support, investigation and supervision were provided by H.-R.Yoon.

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Correspondence to Wonjae Lee or Hye-Ran Yoon.

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Lee, J., Adhikari, S., Lee, W. et al. Development of Ultra High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Enantiomer Resolution of Thyroxine on a Chiral Crown Ether Derived Chiral Stationary Phase. Chromatographia 86, 13–20 (2023). https://doi.org/10.1007/s10337-022-04219-y

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  • DOI: https://doi.org/10.1007/s10337-022-04219-y

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