PuSH - Publication Server of Helmholtz Zentrum München: Determination of 3-iodothyronamine (3-T<sub>1</sub>AM) in mouse liver using liquid chromatography-tandem mass spectrometry.

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Li, Z.M. ; Miller, M. ; Gachkar, S.* ; Mittag, J.* ; Schriever, S.C. ; Pfluger, P.T. ; Schramm, K.-W. ; de Angelis, M.

Determination of 3-iodothyronamine (3-T₁AM) in mouse liver using liquid chromatography-tandem mass spectrometry.

J. Chromatogr. B 1165:122553 (2021)

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3-iodothyronamine (3-T1AM) has been suggested as a novel chemical messenger and potent trace amine-associated receptor 1 ligand in the CNS that occurs naturally as endogenous metabolite of the thyroid hormones. Discrepancies and variations in 3-T1AM plasma and tissue concentrations have nonetheless caused controversy regarding the existence and biological role of 3-T1AM. These discussions are at least partially based on potential analytical artefacts caused by differential decay kinetics of 3-T1AM and the widely used deuterated quantification standard D4-T1AM. Here, we report a novel LC-MS/MS method for the quantification of 3-T1AM in biological specimens using stable isotope dilution with 13C6-T1AM, a new internal standard that showed pharmacodynamic properties comparable to endogenous 3-T1AM. The method detection limit (MDL) and method quantification limit (MQL) of 3-T1AM were 0.04 and 0.09 ng/g, respectively. The spike-recoveries of 3-T1AM were between 85.4% and 94.3%, with a coefficient of variation of 3.7–5.8%. The intra-day and inter-day variations of 3-T1AM were 8.45–11.2% and 3.58–5.73%, respectively. Endogenous 3-T1AM liver values in C57BL/6J mice were 2.20 ± 0.49 pmol/g with a detection frequency of 50%. Higher liver 3-T1AM values were found when C57BL/6J mice were treated with N-acetyl-3-iodothyronamine or O-acetyl-3-iodothyronamine. Overall, our new stable isotope dilution LC-MS/MS method improves both the sensitivity and selectivity compared with existing methods. The concomitant possibility to quantify additional thyroid hormones such as thyroxine, 3,5,3′-triiodo-L-thyronine, 3,3′,5′-triiodo-L-thyronine, 3,3′-diiodo-L-thyronine, and 3,5-diiodo-L-thyronine further adds to the value of our novel method in exploring the natural occurrence and fate of 3-T1AM in biological tissues and fluids.

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PuSH - Publication Server of Helmholtz Zentrum München

Determination of 3-iodothyronamine (3-T1AM) in mouse liver using liquid chromatography-tandem mass spectrometry.

Determination of 3-iodothyronamine (3-T₁AM) in mouse liver using liquid chromatography-tandem mass spectrometry.