Determination of 3-Nitrotyrosine by High-Pressure Liquid Chromatography with a Dual-Mode Electrochemical Detector

Sodum, Rama S.; Akerkar, Shobha A.; Fiala, Emerich S.

doi:10.1006/abio.2000.4528

Cited by 53 publications

(40 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, nitrotyrosine formation is accepted as a biochemical marker for ONOO-(peroxynitrite) formation [58,59]. In our experiments, we were able to observe a significant increase of nitrotyrosine formation after Pb exposure [43], pointed out that nitrotyrosine; the footprint of NO oxidation by ROS, was significantly increased in plasma, kidney, heart, liver, and brain of Pb exposed rats.…”

Section: Pb Increased Ros Productionsupporting

confidence: 60%

Serum c-kit Protein Detection as a Reliable Biomarker for the Diagnosis of Gastrointestinal Stromal Tumors: a Case-Control Study

Apostolopoulos¹

2014

J Mol Biomark Diagn

View full text Add to dashboard Cite

Lead (Pb) is one of the most important environmental pollutant metals accumulating in the atmosphere, water, foods, and in organisms living in contaminated areas. Skin is one of the main targets of Pb toxicity based on its ability of direct penetration upon exposure. The underlying cell damaging pathomechanisms have not been revealed in detail. Herein, we focus on Pb-induced oxidative and nitrosative stress that has not been previously thoroughly investigated. We investigated these effects in order to elucidate the pathomechanisms and as well to identify potential biological markers that may indicate Pb toxicity. Human immortalized keratinocytes (HaCaT cells) were exposed to Pb (100 µM) either for 5 minutes or 6 hours. Pb-induced cellular damage was evaluated by immunocytochemistry analysis of multiple signalling cascades, e.g. apoptosis, Akt, MAPK, NOS, nitrotyrosine and 8-isoprostane formation, detection of nitrosative stress using Diaminofluorescein (DAF-FM) and oxidative stress using 3'-(p aminophenyl) fluorescein (APF). We found that Pb exposure resulted in significantly enhanced NO and ROS production in HaCaT cells. Pb led to enhanced eNOS-phosphorylation at Ser 1177 , and Ser 116 residues but not Thr 495 . AKT phosphorylation but not MAP kinases were enhanced by Pb In addition, Pb induced apoptosis as shown by Caspase-3 activation and PARP cleavage. Our results suggest that Pb mediates its toxic effect in keratinocytes through oxidative and nitrosative stress which is accompanied by differential changes of eNOS phosphorylation and apoptosis. These data significantly contribute to understanding of underlying mechanisms of Pb-induced cellular damage.

show abstract

Section: Pb Increased Ros Productionsupporting

confidence: 60%

Serum c-kit Protein Detection as a Reliable Biomarker for the Diagnosis of Gastrointestinal Stromal Tumors: a Case-Control Study

Apostolopoulos¹

2014

J Mol Biomark Diagn

View full text Add to dashboard Cite

show abstract

“…The results showed that the detection limit of the present assay, at less than 10 fmol, to be remarkably sensitive. Its sensitivity was better than that reported by Sodum et al [10], and continuous measurement was possible. Its detection specificity was also assessed by examining the effects of reduced voltage on peaks.…”

Section: Detection Of 3-nitrotyrosinementioning

confidence: 63%

“…Among the studies on 3-nitrotyrosine detection utilizing this method [3,7,9,10], the dual-mode electrochemical detection method reported by Sodum et al [10] has the highest sensitivity. However, with their method, responses tend to change the mercury surface periodically and redraw the calibration curve.…”

mentioning

confidence: 99%

Determination of Nitrotyrosine by HPLC-ECD and Its Application.

Ishida

Hasegawa

Mukai

et al. 2002

The Journal of Veterinary Medical Science

View full text Add to dashboard Cite

ABSTRACT. 3-nitrotyrosine, a product of tyrosine nitration, is a useful indicator of oxidative damage. We modified the previously reported HPLC-electrochemical detection (ECD) method: specifically, a through-type porous carbon electrode was used as a reducing electrode instead of the mercury-gold amalgam electrode, because the response of the latter changes over time. A combination of reverse-phase HPLC and electrochemical detector passed through -800 mV reduction potential and subsequently under +250 mV oxidation potential allows measurement of 3-nitrotyrosine. The detection limit of this assay was less than 10 fmol. In mice to which lipopolysaccharide (LPS) was administered intraperitoneally, plasma 3-nitrotyrosine levels were elevated, corresponding to LPS dosage. These findings suggest that the improved HPLC-ECD method can be used as a specific and sensitive assay of biological 3-nitrotyrosine and can be applied clinically. KEY WORDS: HPLC-ECD, LPS, 3-nitrotyrosine, peroxynitrite.

show abstract

“…Initially, we tried to chemically (using dithionite, borohydride, or Sn-borohydride under phase transfer conditions) or electrochemically reduce NGT to 5-amino-␥T, a strategy previously employed for the detection of 3-nitrotyrosine (11,21,22), since nitration is known to increase the oxidation potential of phenolic compounds, and aminophenols are much more electrochemically active. However, none of these approaches nor UV photolysis (21) was successful.…”

Section: Hplc Separation and Electrochemical Detectionmentioning

confidence: 99%

Analysis of plasma tocopherols α, γ, and 5-nitro-γ in rats with inflammation by HPLC coulometric detection

Christen

Jiang

Shigenaga

et al. 2002

Journal of Lipid Research

View full text Add to dashboard Cite

Reactive nitrogen oxide species (RNOS) have been implicated as effector molecules in inflammatory diseases. There is emerging evidence that ␥ -tocopherol ( ␥ T), the major form of vitamin E in the North American diet, may play an important role in these diseases. ␥ T scavenges RNOS such as peroxynitrite by forming a stable adduct, 5-nitro-␥ T (NGT). Here we describe a convenient HPLC method for the simultaneous determination of NGT, ␣ T, and ␥ T in blood plasma and other tissues. Coulometric detection of NGT separated on a deactivated reversed-phase column was linear over a wide range of concentrations and highly sensitive ( ‫ف‬ 10 fmol detection limit). NGT extracted from blood plasma of 15-week-old Fischer 344 rats was in the low nM range, representing ‫ف‬ 4% of ␥ T. Twenty-four h after intraperitoneal injection of zymosan, plasma NGT levels were 2-fold higher compared to fasted control animals when adjusted to ␥ T or corrected for total neutral lipids, while ␣ -and ␥ T levels remained unchanged. These results demonstrate that nitration of ␥ T is increased under inflammatory conditions and highlight the importance of RNOS reactions in the lipid phase. The present HPLC method should be helpful in clarifying the precise physiological role of ␥ T. Reactive nitrogen oxide species (RNOS) have been implicated as important effector molecules in inflammatory diseases (1, 2). On the other hand, antioxidants such as vitamin E have been proposed to modulate the effects mediated by RNOS and thus provide protection against inflammation-induced pathology.Vitamin E is a family of lipid-soluble chain-braking antioxidants that share a similar chromanol ring structure ( Fig.

show abstract

Determination of 3-Nitrotyrosine by High-Pressure Liquid Chromatography with a Dual-Mode Electrochemical Detector

Cited by 53 publications

References 34 publications

Serum c-kit Protein Detection as a Reliable Biomarker for the Diagnosis of Gastrointestinal Stromal Tumors: a Case-Control Study

Serum c-kit Protein Detection as a Reliable Biomarker for the Diagnosis of Gastrointestinal Stromal Tumors: a Case-Control Study

Determination of Nitrotyrosine by HPLC-ECD and Its Application.

Analysis of plasma tocopherols α, γ, and 5-nitro-γ in rats with inflammation by HPLC coulometric detection

Contact Info

Product

Resources

About