Hydroxylation of 3-nitrotyrosine （3-NT） and 3-NT containing peptide Gly-nitroTyr-Gly in aqueous solution by hydroxyl radical were investigated with gamma irradiation. The structures of the hydroxylated products were...Hydroxylation of 3-nitrotyrosine （3-NT） and 3-NT containing peptide Gly-nitroTyr-Gly in aqueous solution by hydroxyl radical were investigated with gamma irradiation. The structures of the hydroxylated products were confirmed by electrospray ionization mass spectrometry and ^1H NMR spectrometry. The reactivity of 3-nitrotyrosine has been investigated using density functional theory （DFT） calculation.展开更多
Objective: Myeloperoxidase (MPO) is an inflammatory enzyme that is mainly released by activated neutrophils and monocytes. 3-nitrotyrosine (NT) is a stable inflammatory end product of MPO that is produced through nitr...Objective: Myeloperoxidase (MPO) is an inflammatory enzyme that is mainly released by activated neutrophils and monocytes. 3-nitrotyrosine (NT) is a stable inflammatory end product of MPO that is produced through nitrosylation of free and protein-bound tyrosines. Determination of the exact levels of free NT is technically a challenging matter. Also, there is limited information about the relationship between MPO and free NT levels and elevation of them in the plasma of patients with coronary artery disease (CAD). Therefore, we sought to determine the exact level of plasma free NT with a simple and exquisite technique in CAD patients. Methods: This study included 50 stable angina, 50 unstable angina patients, and 50 control subjects. Plasma MPO concentration was measured with an immunoassay method. Plasma free NT level was determined by a modified HPLC-fluorescence method. Lipid profile, high sensitivity C-reactive protein (hsCRP) and other clinical risk factors of patients were also assigned. Results: Plasma level of free NT was efficiently measured by the HPLC-fluorescence method. Plasma levels of MPO and NT were significantly higher in patients with stable and unstable CAD than in control subjects (P < 0.001). There was a significant correlation between the two substances in CAD patients (P < 0.001). Conclusions: We determined plasma free NT levels with a sensitive HPLC-fluorescence method with some modifications in a clinical scale. Plasma levels of MPO and NT were profoundly elevated in CAD patients. The significant relationships of the two substances and elevation of them may have useful clinical implication in patients with stable and unstable CAD.展开更多
A novel method for the simultaneous determination of 3-nitrotyrosine(NT) and 3-chlorotyrosine(CT) in human plasma has been developed based on direct analysis in real time–tandem mass spectrometry(DART–MS/MS). Analys...A novel method for the simultaneous determination of 3-nitrotyrosine(NT) and 3-chlorotyrosine(CT) in human plasma has been developed based on direct analysis in real time–tandem mass spectrometry(DART–MS/MS). Analysis was performed in the positive ionization mode using multiple reaction monitoring(MRM) of the ion transitions at m/z 216.2/170.1 for CT, m/z 227.2/181.1 for NT and m/z 230.2/184.2 for the internal standard, d^3-NT. The assay was linear in the ranges 0.5–100 μg/m L for CT and 4–100 μg/m L for NT with corresponding limits of detection of 0.2 and 2 μg/m L. Intra- and inter-day precisions and accuracies were respectively <15% and ±15%. Matrix effects were also evaluated. The method is potentially useful for high throughput analysis although sensitivity needs to be improved before it can be applied in clinical research.展开更多
基金The authors would like to thank the financial support from the National Natural Science Foundation of China （No. 20320130046） ；the Specialized Research Fund for the Doctoral Program of Higher Education （SRFDP） of M0E （No. 20030003049）.
文摘Hydroxylation of 3-nitrotyrosine （3-NT） and 3-NT containing peptide Gly-nitroTyr-Gly in aqueous solution by hydroxyl radical were investigated with gamma irradiation. The structures of the hydroxylated products were confirmed by electrospray ionization mass spectrometry and ^1H NMR spectrometry. The reactivity of 3-nitrotyrosine has been investigated using density functional theory （DFT） calculation.
文摘Objective: Myeloperoxidase (MPO) is an inflammatory enzyme that is mainly released by activated neutrophils and monocytes. 3-nitrotyrosine (NT) is a stable inflammatory end product of MPO that is produced through nitrosylation of free and protein-bound tyrosines. Determination of the exact levels of free NT is technically a challenging matter. Also, there is limited information about the relationship between MPO and free NT levels and elevation of them in the plasma of patients with coronary artery disease (CAD). Therefore, we sought to determine the exact level of plasma free NT with a simple and exquisite technique in CAD patients. Methods: This study included 50 stable angina, 50 unstable angina patients, and 50 control subjects. Plasma MPO concentration was measured with an immunoassay method. Plasma free NT level was determined by a modified HPLC-fluorescence method. Lipid profile, high sensitivity C-reactive protein (hsCRP) and other clinical risk factors of patients were also assigned. Results: Plasma level of free NT was efficiently measured by the HPLC-fluorescence method. Plasma levels of MPO and NT were significantly higher in patients with stable and unstable CAD than in control subjects (P < 0.001). There was a significant correlation between the two substances in CAD patients (P < 0.001). Conclusions: We determined plasma free NT levels with a sensitive HPLC-fluorescence method with some modifications in a clinical scale. Plasma levels of MPO and NT were profoundly elevated in CAD patients. The significant relationships of the two substances and elevation of them may have useful clinical implication in patients with stable and unstable CAD.
文摘A novel method for the simultaneous determination of 3-nitrotyrosine(NT) and 3-chlorotyrosine(CT) in human plasma has been developed based on direct analysis in real time–tandem mass spectrometry(DART–MS/MS). Analysis was performed in the positive ionization mode using multiple reaction monitoring(MRM) of the ion transitions at m/z 216.2/170.1 for CT, m/z 227.2/181.1 for NT and m/z 230.2/184.2 for the internal standard, d^3-NT. The assay was linear in the ranges 0.5–100 μg/m L for CT and 4–100 μg/m L for NT with corresponding limits of detection of 0.2 and 2 μg/m L. Intra- and inter-day precisions and accuracies were respectively <15% and ±15%. Matrix effects were also evaluated. The method is potentially useful for high throughput analysis although sensitivity needs to be improved before it can be applied in clinical research.
