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Ratiometric quantitation of thiol metabolites using non-isotopic mass tags.

Analytica chimica acta (2018-10-08)
Xiaofeng Zhao, Dawn S Hui, Richard Lee, James L Edwards
ANOTACE

Ratiometric quantitation is used in mass spectrometry to account for variations in ionization efficiencies due to heterogenous sample matrixes. Isotopes are most commonly used to achieve ratiometric quantitation because of their ability to co-elute chromatographically with each other and to have similar ionization efficiencies. In the work presented here, a new non-isotopic quantitative tagging approach is presented which allows chromatographic co-elution and similar ionization efficiencies. Using two variations of maleimide tags, t-butyl and cyclohexyl maleimide, thiols are quantified with a high degree of linearity up to five-fold concentration differences. Because these two tags have similar hydrophobcities, they elute simultaneously which allows them to be used for ratiometric quantitation. Beyond the five-fold linear range, signal compression is observed. This technique was able to quantify thiol changes in both in vitro pharmacological treatments as well as in vivo diabetic tissue.

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Sigma-Aldrich
Cys-Gly, ≥85% (TLC)
Sigma-Aldrich
DL-Cysteine, technical grade
Sigma-Aldrich
N-tert-Butylmaleimide, 97%