HPLC study of oxidation products of hydroethidine in chemical and biological systems: ramifications in superoxide measurements

被引：124

作者：

Zielonka, Jacek ^{[1
,2
]}

Hardy, Micael ^{[3
,4
,5
,6
]}

Kalyanaraman, B. ^{[1
,2
]}

机构：

[1] Med Coll Wisconsin, Dept Biophys, Milwaukee, WI 53226 USA

[2] Med Coll Wisconsin, Free Rad Res Ctr, Milwaukee, WI 53226 USA

[3] CNRS, UMR 6517, Lab SREP, F-13397 Marseille, France

[4] Univ Aix Marseille 1, Ctr St Jerome, F-13397 Marseille, France

[5] Univ Aix Marseille 2, Ctr St Jerome, F-13397 Marseille, France

[6] Univ Aix Marseille 3, Ctr St Jerome, F-13397 Marseille, France

来源：

FREE RADICAL BIOLOGY AND MEDICINE | 2009年 / 46卷 / 03期

基金：

美国国家卫生研究院;

关键词：

Hydroethidine; Mito-hydroethidine; MitoSOX; HPLC; Superoxide; Fluorescence; Ethidium; Diethidium; 2-Hydroxyethidium; Free radicals; NITRIC-OXIDE SYNTHASE; FLUORESCENCE DETECTION; INTRACELLULAR SUPEROXIDE; ENDOTHELIAL-CELLS; HYDROGEN-PEROXIDE; HYPERTENSION; GENERATION; DISMUTASE; ETHIDIUM; PROBES;

D O I：

10.1016/j.freeradbiomed.2008.10.031

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Methods for the detection and quantitation of hydroethidine (HE) and its oxidation products by HPLC analysis are described. Synthetic methods for preparation of authentic standards (2-hydroxyethidium and diethidium) are provided. Potential applications of the HPLC methods to chemical and biological systems are discussed. Specific examples of chromatograms obtained using UV-Vis absorption, fluorescence, electrochemical, and mass spectrometry detectors are provided. The development of a dual electrochemical and fluorescence detection methodology and its applications are described. The HPLC-based method enables analyses of HE and its oxidation products such as ethidium and the dimeric products of HE. The ramifications of HPLC measurement of HE and its oxidation products in the detection and quantitation of 2-hydroxyethidium, the diagnostic market product of superoxide and HE, in the intracellular milieu are discussed. Similarly, mitochondria-targeted HE conjugated to a triphenylphosphonium group (Mito-HE or Mito-SOX) also forms oxidation products (dimers of Mito-HE and Mito-E+) that can affect the detection and quantitation of 2-hydroxy-mito-ethidium, the diagnostic market product of Mito-HE and superoxide in mitochondria. (C) 2008 Elsevier Inc. All rights reserved.

引用

页码：329 / 338

页数：10

共 47 条

[1] Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa
Aitken, R. John
Wingate, Jordana K.
De Iuliis, Geoffry N.
Koppers, Adam J.
McLaughlin, Eileen A.
[J]. JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 2006, 91 (10) : 4154 - 4163
[2] Glycolaldehyde induces growth inhibition and oxidative stress in human breast cancer cells
Al-Enezi, KS
Alkhalaf, M
Benov, LT
[J]. FREE RADICAL BIOLOGY AND MEDICINE, 2006, 40 (07) : 1144 - 1151
[3] Critical evaluation of the use of hydroethidine as a measure of superoxide anion radical
Benov, L
Sztejnberg, L
Fridovich, I
[J]. FREE RADICAL BIOLOGY AND MEDICINE, 1998, 25 (07) : 826 - 831
[4] Angiotensin II chronic infusion induces B1 receptor expression in aorta of rats
Ceravolo, Graziela S.
Fernandes, Liliam
Munhoz, Carolina D.
Fernandes, Denise C.
Tostes, Rita C. A.
Laurindo, Francisco R. M.
Scavone, Christoforo
Fortes, Zuleica B.
Carvalho, Maria Helena C.
[J]. HYPERTENSION, 2007, 50 (04) : 756 - 761
[5] A selective, cell-permeable optical probe for hydrogen peroxide in living cells
Chang, MCY
Pralle, A
Isacoff, EY
Chang, CJ
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (47) : 15392 - 15393
[6] S-glutathiolation of p21ras by peroxynitrite mediates endothelial insulin resistance caused by oxidized low-density lipoprotein
Clavreul, Nicolas
Bachschmid, Markus M.
Hou, Xiuyun
Shi, Chaomei
Idrizovic, Azra
Ido, Yasuo
Pimentel, David
Cohen, Richard A.
[J]. ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2006, 26 (11) : 2454 - 2461
[7] Definitive evidence for the nonmitochondrial production of superoxide anion by human spermatozoa
De Iuliis, GN
Wingate, JK
Koppers, AJ
McLaughlin, EA
Aitken, RJ
[J]. JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 2006, 91 (05) : 1968 - 1975
[8] A targetable fluorescent probe for imaging hydrogen peroxide in the mitochondria of living cells
Dickinson, Bryan C.
Chang, Christopher J.
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2008, 130 (30) : 9638 - +
[9] Measurement of reactive oxygen species in cardiovascular studies
Dikalov, Sergey
Griendling, Kathy K.
Harrison, David G.
[J]. HYPERTENSION, 2007, 49 (04) : 717 - 727
[10] Molecular mechanisms of angiotensin II-mediated mitochondrial dysfunction - Linking mitochondrial oxidative damage and vascular endothelial dysfunction
Doughan, Abdulrahman K.
Harrison, David G.
Dikalov, Sergey I.
[J]. CIRCULATION RESEARCH, 2008, 102 (04) : 488 - 496

← 1 2 3 4 5 →