Abstract
The transfer of a gradient method to an isocratic or multistep gradient method employing stationary phase optimized liquid chromatography facilitated a reduction in analysis time by 50% and significantly improved the mass spectrometric detectability of impurities in synthetic thyroid hormones. Four column segments packed with different stationary phases were combined into a single chromatographic column, which allowed the separation and photometric as well as mass spectrometric detection of thyroid compounds in less than 30 min under isocratic- or step gradient elution conditions with 0.10% acetic acid/acetonitrile. Signal instability and baseline drift during detection by negative electrospray ionization time-of-flight mass spectrometry were minimized by optimizing the spray parameters for each individual elution step. This resulted in improved detectabilities and higher mass spectral quality, especially for low-abundance components in the sample mixture. The method was applied to the separation and detection of the low-abundance impurities formed upon the thermal stressing of a sample of synthetic levothyroxine.
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Financial support by Bischoff Analysentechnik und -geräte GmbH (Leonberg, Germany) and helpful discussions with K. Bischoff and S. Lamotte are gratefully acknowledged.
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Gostomski, I., Braun, R. & Huber, C.G. Detection of low-abundance impurities in synthetic thyroid hormones by stationary phase optimized liquid chromatography–mass spectrometry. Anal Bioanal Chem 391, 279–288 (2008). https://doi.org/10.1007/s00216-008-1920-4
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DOI: https://doi.org/10.1007/s00216-008-1920-4