Synthesis of trityl radical-conjugated disulfide biradicals for measurement of thiol concentration

Yangping Liu, Yuguang Song, A. Rockenbauer, Jian Sun, Craig Hemann, Frederick A. Villamena, Jay L. Zweier

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Measurement of thiol concentrations is of great importance for characterizing their critical role in normal metabolism and disease. Low-frequency electron paramagnetic resonance (EPR) spectroscopy and imaging, coupled with the use of exogenous paramagnetic probes, have been indispensable techniques for the in vivo measurement of various physiological parameters owing to the specificity, noninvasiveness and good depth of magnetic field penetration in animal tissues. However, in vivo detection of thiol levels by EPR spectroscopy and imaging is limited due to the need for improved probes. We report the first synthesis of trityl radical-conjugated disulfide biradicals (TSSN and TSST) as paramagnetic thiol probes. The use of trityl radicals in the construction of these biradicals greatly facilitates thiol measurement by EPR spectroscopy since trityls have extraordinary stability in living tissues with a single narrow EPR line that enables high sensitivity and resolution for in vivo EPR spectroscopy and imaging. Both biradicals exhibit broad characteristic EPR spectra at room temperature because of their intramolecular spin-spin interaction. Reaction of these biradicals with thiol compounds such as glutathione (GSH) and cysteine results in the formation of trityl monoradicals which exhibit high spectral sensitivity to oxygen. The moderately slow reaction between the biradicals and GSH (k2 ∼ 0.3 M-1 s -1 for TSSN and 0.2 M-1 s-1 for TSST) allows for in vivo measurement of GSH concentration without altering the redox environment in biological systems. The GSH concentration in rat liver was determined to be 3.49 ± 0.14 mM by TSSN and 3.67 ± 0.24 mM by TSST, consistent with the value (3.71 ± 0.09 mM) determined by the Ellmans reagent. Thus, these trityl-based thiol probes exhibit unique properties enabling measurement of thiols in biological systems and should be of great value for monitoring redox metabolism.

Original languageEnglish
Pages (from-to)3853-3860
Number of pages8
JournalJournal of Organic Chemistry
Volume76
Issue number10
DOIs
Publication statusPublished - May 20 2011

Fingerprint

Sulfhydryl Compounds
Disulfides
Paramagnetic resonance
Spectroscopy
Biological systems
Imaging techniques
Metabolism
Tissue
Dithionitrobenzoic Acid
Liver
Glutathione
Cysteine
Rats
Animals
Magnetic fields
Oxygen
Monitoring

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Synthesis of trityl radical-conjugated disulfide biradicals for measurement of thiol concentration. / Liu, Yangping; Song, Yuguang; Rockenbauer, A.; Sun, Jian; Hemann, Craig; Villamena, Frederick A.; Zweier, Jay L.

In: Journal of Organic Chemistry, Vol. 76, No. 10, 20.05.2011, p. 3853-3860.

Research output: Contribution to journalArticle

Liu, Yangping ; Song, Yuguang ; Rockenbauer, A. ; Sun, Jian ; Hemann, Craig ; Villamena, Frederick A. ; Zweier, Jay L. / Synthesis of trityl radical-conjugated disulfide biradicals for measurement of thiol concentration. In: Journal of Organic Chemistry. 2011 ; Vol. 76, No. 10. pp. 3853-3860.
@article{c499c61a5f4c498abfe4ee6e5b1a8dae,
title = "Synthesis of trityl radical-conjugated disulfide biradicals for measurement of thiol concentration",
abstract = "Measurement of thiol concentrations is of great importance for characterizing their critical role in normal metabolism and disease. Low-frequency electron paramagnetic resonance (EPR) spectroscopy and imaging, coupled with the use of exogenous paramagnetic probes, have been indispensable techniques for the in vivo measurement of various physiological parameters owing to the specificity, noninvasiveness and good depth of magnetic field penetration in animal tissues. However, in vivo detection of thiol levels by EPR spectroscopy and imaging is limited due to the need for improved probes. We report the first synthesis of trityl radical-conjugated disulfide biradicals (TSSN and TSST) as paramagnetic thiol probes. The use of trityl radicals in the construction of these biradicals greatly facilitates thiol measurement by EPR spectroscopy since trityls have extraordinary stability in living tissues with a single narrow EPR line that enables high sensitivity and resolution for in vivo EPR spectroscopy and imaging. Both biradicals exhibit broad characteristic EPR spectra at room temperature because of their intramolecular spin-spin interaction. Reaction of these biradicals with thiol compounds such as glutathione (GSH) and cysteine results in the formation of trityl monoradicals which exhibit high spectral sensitivity to oxygen. The moderately slow reaction between the biradicals and GSH (k2 ∼ 0.3 M-1 s -1 for TSSN and 0.2 M-1 s-1 for TSST) allows for in vivo measurement of GSH concentration without altering the redox environment in biological systems. The GSH concentration in rat liver was determined to be 3.49 ± 0.14 mM by TSSN and 3.67 ± 0.24 mM by TSST, consistent with the value (3.71 ± 0.09 mM) determined by the Ellmans reagent. Thus, these trityl-based thiol probes exhibit unique properties enabling measurement of thiols in biological systems and should be of great value for monitoring redox metabolism.",
author = "Yangping Liu and Yuguang Song and A. Rockenbauer and Jian Sun and Craig Hemann and Villamena, {Frederick A.} and Zweier, {Jay L.}",
year = "2011",
month = "5",
day = "20",
doi = "10.1021/jo200265u",
language = "English",
volume = "76",
pages = "3853--3860",
journal = "Journal of Organic Chemistry",
issn = "0022-3263",
publisher = "American Chemical Society",
number = "10",

}

TY - JOUR

T1 - Synthesis of trityl radical-conjugated disulfide biradicals for measurement of thiol concentration

AU - Liu, Yangping

AU - Song, Yuguang

AU - Rockenbauer, A.

AU - Sun, Jian

AU - Hemann, Craig

AU - Villamena, Frederick A.

AU - Zweier, Jay L.

PY - 2011/5/20

Y1 - 2011/5/20

N2 - Measurement of thiol concentrations is of great importance for characterizing their critical role in normal metabolism and disease. Low-frequency electron paramagnetic resonance (EPR) spectroscopy and imaging, coupled with the use of exogenous paramagnetic probes, have been indispensable techniques for the in vivo measurement of various physiological parameters owing to the specificity, noninvasiveness and good depth of magnetic field penetration in animal tissues. However, in vivo detection of thiol levels by EPR spectroscopy and imaging is limited due to the need for improved probes. We report the first synthesis of trityl radical-conjugated disulfide biradicals (TSSN and TSST) as paramagnetic thiol probes. The use of trityl radicals in the construction of these biradicals greatly facilitates thiol measurement by EPR spectroscopy since trityls have extraordinary stability in living tissues with a single narrow EPR line that enables high sensitivity and resolution for in vivo EPR spectroscopy and imaging. Both biradicals exhibit broad characteristic EPR spectra at room temperature because of their intramolecular spin-spin interaction. Reaction of these biradicals with thiol compounds such as glutathione (GSH) and cysteine results in the formation of trityl monoradicals which exhibit high spectral sensitivity to oxygen. The moderately slow reaction between the biradicals and GSH (k2 ∼ 0.3 M-1 s -1 for TSSN and 0.2 M-1 s-1 for TSST) allows for in vivo measurement of GSH concentration without altering the redox environment in biological systems. The GSH concentration in rat liver was determined to be 3.49 ± 0.14 mM by TSSN and 3.67 ± 0.24 mM by TSST, consistent with the value (3.71 ± 0.09 mM) determined by the Ellmans reagent. Thus, these trityl-based thiol probes exhibit unique properties enabling measurement of thiols in biological systems and should be of great value for monitoring redox metabolism.

AB - Measurement of thiol concentrations is of great importance for characterizing their critical role in normal metabolism and disease. Low-frequency electron paramagnetic resonance (EPR) spectroscopy and imaging, coupled with the use of exogenous paramagnetic probes, have been indispensable techniques for the in vivo measurement of various physiological parameters owing to the specificity, noninvasiveness and good depth of magnetic field penetration in animal tissues. However, in vivo detection of thiol levels by EPR spectroscopy and imaging is limited due to the need for improved probes. We report the first synthesis of trityl radical-conjugated disulfide biradicals (TSSN and TSST) as paramagnetic thiol probes. The use of trityl radicals in the construction of these biradicals greatly facilitates thiol measurement by EPR spectroscopy since trityls have extraordinary stability in living tissues with a single narrow EPR line that enables high sensitivity and resolution for in vivo EPR spectroscopy and imaging. Both biradicals exhibit broad characteristic EPR spectra at room temperature because of their intramolecular spin-spin interaction. Reaction of these biradicals with thiol compounds such as glutathione (GSH) and cysteine results in the formation of trityl monoradicals which exhibit high spectral sensitivity to oxygen. The moderately slow reaction between the biradicals and GSH (k2 ∼ 0.3 M-1 s -1 for TSSN and 0.2 M-1 s-1 for TSST) allows for in vivo measurement of GSH concentration without altering the redox environment in biological systems. The GSH concentration in rat liver was determined to be 3.49 ± 0.14 mM by TSSN and 3.67 ± 0.24 mM by TSST, consistent with the value (3.71 ± 0.09 mM) determined by the Ellmans reagent. Thus, these trityl-based thiol probes exhibit unique properties enabling measurement of thiols in biological systems and should be of great value for monitoring redox metabolism.

UR - http://www.scopus.com/inward/record.url?scp=79956136766&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79956136766&partnerID=8YFLogxK

U2 - 10.1021/jo200265u

DO - 10.1021/jo200265u

M3 - Article

C2 - 21488696

AN - SCOPUS:79956136766

VL - 76

SP - 3853

EP - 3860

JO - Journal of Organic Chemistry

JF - Journal of Organic Chemistry

SN - 0022-3263

IS - 10

ER -