2.2 Biochemistry analysis
2.2.1 Peroxide assay. The peroxide content was determined in
plasma by means of a quantitative colorimetric assay, according to the
kit indications ”QuantiChromTM Peroxide Assay Kit
(DIOX-250) - BioAssay Systems”, at the end of the process the optical
density at 570 nm was quantified in ELISA plate reading equipment
(Chromate), reporting values of ng/ml.
2.2.2 Nitric
oxide assay. The nitric oxide content was determined in plasma by means
of a quantitative colorimetric assay, according to the kit indications
”QuantiChromTM Nitric Oxide Assay Kit (D2NO-100) -
BioAssay Systems”, at the end of the process the optical density was
quantified at 570 nm, in an ELISA plate reading equipment (Chromate),
reporting in µM values.
2.2.3 Oxidized proteins assay. Protein carbonyl content in
plasma was determined as previously described.12 The
plasma was incubated with DNPH (2,4-Dinitrophenylhydrazine; 10 mM) and
HCl (2N) and finally with guanidine hydrochloride (6M). Assessment of
carbonyl formation was done on the basis of formation of protein
hydrazone by reaction with DNPH. The absorbance was measured at 370 nm.
Result was expressed as nmol carbonyl per mg protein.