Measurement and Clinical Significance of Biomarkers of Oxidative Stress in Humans

Table 2

Markers based on ROS-induced modifications.

Markers	Methods	Limitations and confoundings
Lipid oxidation
HNE	HPLC, GC-MS Immunoassay
MDA, alkenals, alkadienals	Spectrophotometric/fluorimetric (TBARS), HPLC (UV or fluorescence)Immunoassay	Sugars, amino acids, bilirubin and albumin, hemolysis
F2-IsoPs	Gas/liquid chromatography coupled with mass spectroscopy techniques Immunoassay	Hemolysis Antibody specificity
DNA oxidation
8oxodG, 5-chlorocytosine, 5-chlorouracil, εdA, εdC	ELISA assays, HPLC-ECD, HPLC/GC-MS	Antibody specificity
Protein oxidation
ALEs, AGEs	HPLC, Western blot after one-dimensional or two-dimensional electrophoretic separation, immunohistochemistry, ELISA	Structural heterogeneity of these products Antibody specificity
Carbonils	Spectrophotometric, HPLC, ELISA
3-NO-Tyr	HPLC/GC-MS, ELISA	Possible nitration of tyrosine residues in the sample by the presence of nitrite and the acid conditions during protein precipitation and hydrolysis Antibody specificity
AOPP	MS, colorimetric assays
oxLDL	Immunodetection (ELISA)	Antibody specificity
IMA	ABC test, immunodetection (ELISA)	Sensitive to pH changes, temperature, and time of sample storage Antibody specificity

8oxodG: 7,8-dihydroxy-8-oxo-2′-deoxyguanosine; ABC test: binding capacity of albumin for cobalt; AGEs: advanced glycation end products; ALEs: advanced lipoxigenation end products; AOPP: advanced oxidation protein products; F2-IsoPs: F2-isoprostanes; GC: gas chromatography; HNE: 4-hydroxy-2-nonenal; HPLC: high-performance liquid chromatography; ECD: electrochemical detection; IMA: ischemia-modified albumin; MS: mass spectroscopy; MDA: malondialdehyde; TBARS: thiobarbituric acid reactive substances.