Automated Online Solid-Phase Derivatization for Sensitive Quantification of Endogenous S-Nitrosoglutathione and Rapid Capture of Other Low-Molecular-Mass S-Nitrosothiols
| Autor: | John S. Wishnok, Steven R. Tannenbaum, Xin Wang, Carlos T. Garcia, Guanyu Gong |
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| Rok vydání: | 2018 |
| Předmět: |
Analyte
Time Factors Monolithic HPLC column Endogeny 010402 general chemistry Mass spectrometry 01 natural sciences Mass Spectrometry Article Analytical Chemistry S-Nitrosoglutathione chemistry.chemical_compound Limit of Detection Animals Derivatization Chromatography High Pressure Liquid Detection limit S-Nitrosothiols Chromatography Molecular mass Chemistry Solid Phase Extraction 010401 analytical chemistry Equipment Design 0104 chemical sciences Mice Inbred C57BL Molecular Weight Female |
| Zdroj: | Analytical Chemistry. 90:1967-1975 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.7b04049 |
| Popis: | S-Nitrosothiols (RSNOs) constitute a circulating endogenous reservoir of nitric oxide, and have important biological activities. In this study, an online coupling of solid phase derivatization (SPD) with liquid chromatography-mass spectrometry (LC-MS) was developed and applied in the analysis of low-molecular-mass RSNOs. A derivatizing-reagent-modified polymer monolithic column was prepared and adapted for online SPD-LC-MS. Analytes from the LC auto-sampler flowed through the monolithic column for derivatization, and then directly into the LC-MS for analysis. This integration of the online derivatization, LC separation and MS detection facilitated system automation, allowing rapid, laborsaving, and sensitive detection of RSNOs. S-Nitrosoglutathione (GSNO) was quantified using this automated online method with good linearity (R2 = 0.9994); the limit of detection was 0.015 nM. The online SPD-LC-MS method has been used to determine GSNO levels in mouse samples, 138 ± 13.2 nM of endogenous GSNO was detected in mouse plasma. Besides, the GSNO concentrations in liver (64.8 ± 11.3 pmol/mg protein), kidney (47.2 ± 6.1 pmol/mg protein), heart (8.9 ± 1.8 pmol/mg protein), muscle (1.9 ± 0.3 pmol/mg protein), hippocampus (5.3 ± 0.9 pmol/mg protein), striatum (6.7 ± 0.6 pmol/mg protein), cerebellum (31.4 ± 6.5 pmol/mg protein), and cortex (47.9 ± 4.6 pmol/mg protein) were also successfully quantified. When the derivatization was performed within 8 minutes, followed by LC-MS detection, samples could be rapidly analyzed compared with the offline manual method. Other low-molecular-mass RSNOs, such as S-nitrosocysteine and S-nitrosocysteinylglycine, were captured by rapid precursor-ion scanning, showing that the proposed method is a potentially powerful tool for capture, identification and quantification of RSNOs in biological samples. |
| Databáze: | OpenAIRE |
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