Gas Chromatographic–Mass Spectrometric Detection of S-Nitroso-cysteine and S-Nitroso-glutathione
| Autor: | Dimitrios Tsikas, Jürgen C. Frölich, Frank-Mathias Gutzki, Jörg Sandmann, Stefan Rossa |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Erythrocytes
S nitroso glutathione Biophysics Sensitivity and Specificity Biochemistry Gas Chromatography-Mass Spectrometry chemistry.chemical_compound Cytosol Humans Cysteine Nitrite Molecular Biology S-Nitrosothiols Chromatography Nitrogen Isotopes Reproducibility of Results Cell Biology Nitroso Glutathione Mass spectrometric Quantitative determination chemistry Human plasma S-Nitrosoglutathione Human erythrocytes Indicators and Reagents Nitroso Compounds |
| Zdroj: | Analytical Biochemistry. 272:117-122 |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.1999.4177 |
| Popis: | A gas chromatographic-mass spectrometric (GC-MS) method is described for the quantitative determination of the S-nitroso compounds S-nitroso-cysteine (SNC) and S-nitroso-glutathione (GSNO) using their (15)N-labeled analogs, i.e., S(15)NC and GS(15)NO, as internal standards. The method is based on the specific conversion by HgCl(2) of the unlabeled and (15)N-labeled S-nitroso groups to nitrite and (15)N-nitrite, respectively, and their conversion to the pentafluorobenzyl derivatives. The method was applied to quantify GS(15)NO formed in the cytosol of washed human erythrocytes incubated with S(15)NC. Combination of high-performance liquid chromatography with GC-MS allowed specific and accurate quantification of SNC and GSNO externally added to human plasma ultrafiltrate (range 0-10 microM). Method accuracy and precision for SNC and GSNO were close to 100 and below 9%, respectively. As little as 0.1 nM GS(15)NO corresponding to 30 amol of (15)N-nitrite injected onto the column was precisely detected by the method. |
| Databáze: | OpenAIRE |
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