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Isotope dilution‐gas chromatography/mass spectrometry and liquid chromatography/electrospray ionization‐tandem mass spectrometry for the determination of triiodo‐ L ‐thyronine in serum
Author(s) -
Thienpont Linda M.,
Fierens Colette,
De Leenheer André P.,
Przywara Louise
Publication year - 1999
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19991015)13:19<1924::aid-rcm734>3.0.co;2-u
Subject(s) - chemistry , chromatography , isotope dilution , mass spectrometry , electrospray ionization , tandem mass spectrometry , gas chromatography–mass spectrometry , gas chromatography , detection limit , liquid chromatography–mass spectrometry
Isotope dilution‐gas chromatography/mass spectrometry (ID‐GC/MS) and isotope dilution‐liquid chromatography/tandem mass spectrometry (ID‐LC/MS/MS) methods have been developed for an determination of triiodothyronine (T3) in serum and their potential as candidate reference methods investigated. In both methods, 13 C 9 ‐T3 was used as internal standard. Sample pretreatment consisted of deproteinization, extraction and high performance liquid chromatography (HPLC) purification. Conversion of serum thyroxine (T4) to T3 was controlled by adding 13 C 6 ‐T4. For GC/MS, T3 and 13 C 9 ‐T3 were converted to the N,O ‐di‐heptafluorobutyryl (HFB) methyl ester derivatives and monitored at m/z 844 and 853. For LC/MS with electrospray ionization, the transitions m/z 652/661 to 606/614 were monitored. For use of the methods as candidate reference methods, special attention was paid to the calibration and the measurement protocol (duplicate analysis of each sample on three occasions). Evaluation of the ID‐GC/MS and ID‐LC/MS/MS methods showed the absence of interference by reverse T3 and T4, a limit of detection of 100 pg (GC/MS) and 18 pg (LC/MS), a recovery of 100 ± 1.5% (95% confidence interval) and good precision (the total coefficient of variation, n = 6, was typically 1.5%). In addition to the recovery study, the accuracy of the methods was proven by method comparison (ID‐GC/MS vs. LC/MS/MS) on three sera, showing a maximum deviation of 1.1%. Finally, the ID‐GC/MS method was applied for measurement of 10 different human sera with a T3 concentration range from 0.6 to 7.3 ng/mL. Copyright © 1999 John Wiley & Sons, Ltd.