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

Zhong Min Li; Manuel Miller; Sogol Gachkar; Jens Mittag; Sonja C. Schriever; Paul T. Pfluger; Karl Werner Schramm; Meri De Angelis

doi:10.1016/j.jchromb.2021.122553

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

Zhong Min Li^*, Manuel Miller, Sogol Gachkar, Jens Mittag, Sonja C. Schriever, Paul T. Pfluger, Karl Werner Schramm, Meri De Angelis

^*Corresponding author for this work

1 Citation (Scopus)

Abstract

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.

Original language	English
Article number	122553
Journal	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume	1165
ISSN	1570-0232
DOIs	https://doi.org/10.1016/j.jchromb.2021.122553
Publication status	Published - 15.02.2021

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Areas and Centers

Academic Focus: Center for Brain, Behavior and Metabolism (CBBM)

DFG Research Classification Scheme

2.22-17 Endocrinology, Diabetology, Metabolism

Access to Document

10.1016/j.jchromb.2021.122553

Cite this

Li, Z. M., Miller, M., Gachkar, S., Mittag, J., Schriever, S. C., Pfluger, P. T., Schramm, K. W., & De Angelis, M. (2021). Determination of 3-iodothyronamine (3-T₁AM) in mouse liver using liquid chromatography-tandem mass spectrometry. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 1165, Article 122553. https://doi.org/10.1016/j.jchromb.2021.122553

@article{41e38839f4a949f385d7c84207d55712,

title = "Determination of 3-iodothyronamine (3-T1AM) in mouse liver using liquid chromatography-tandem mass spectrometry",

abstract = "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.",

author = "Li, \{Zhong Min\} and Manuel Miller and Sogol Gachkar and Jens Mittag and Schriever, \{Sonja C.\} and Pfluger, \{Paul T.\} and Schramm, \{Karl Werner\} and \{De Angelis\}, Meri",

year = "2021",

month = feb,

day = "15",

doi = "10.1016/j.jchromb.2021.122553",

language = "English",

volume = "1165",

journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",

issn = "1570-0232",

publisher = "Elsevier B.V.",

}

Li, ZM, Miller, M, Gachkar, S, Mittag, J, Schriever, SC, Pfluger, PT, Schramm, KW & De Angelis, M 2021, 'Determination of 3-iodothyronamine (3-T₁AM) in mouse liver using liquid chromatography-tandem mass spectrometry', Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, vol. 1165, 122553. https://doi.org/10.1016/j.jchromb.2021.122553

Determination of 3-iodothyronamine (3-T₁AM) in mouse liver using liquid chromatography-tandem mass spectrometry. / Li, Zhong Min; Miller, Manuel; Gachkar, Sogol et al.
In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 1165, 122553, 15.02.2021.

Research output: Journal Articles › Journal articles › Research › peer-review

TY - JOUR

T1 - Determination of 3-iodothyronamine (3-T1AM) in mouse liver using liquid chromatography-tandem mass spectrometry

AU - Li, Zhong Min

AU - Miller, Manuel

AU - Gachkar, Sogol

AU - Mittag, Jens

AU - Schriever, Sonja C.

AU - Pfluger, Paul T.

AU - Schramm, Karl Werner

AU - De Angelis, Meri

PY - 2021/2/15

Y1 - 2021/2/15

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/85099679854

U2 - 10.1016/j.jchromb.2021.122553

DO - 10.1016/j.jchromb.2021.122553

M3 - Journal articles

AN - SCOPUS:85099679854

SN - 1570-0232

VL - 1165

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

M1 - 122553

ER -

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

Abstract

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DFG Research Classification Scheme

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CRC/TRR 296 LocoTact: Local control of TH action

Cite this

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

Abstract

UN SDGs

Research Areas and Centers

DFG Research Classification Scheme

Access to Document

Other files and links

Fingerprint

Projects

CRC/TRR 296 LocoTact: Local control of TH action

Cite this

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