Abstract
The formation of the hydroxyl free radical (HFR) can be quantified indirectly, by measuring two products of the hydroxylation of salicylic acid, 2,3-dihydroxybenzoate (2,3-DHB) and 2,5-dihydroxybenzoate (2,5-DHB). In this study, we used reversed-phase high-performance liquid chromatography with electrochemical (coulometric) detection to measure 2,3- and 2,5-DHB levels in human platelets. The limits of detection of the method were 10 and 5 fmol on column for 2,3-DHB and 2,5-DHB, respectively. We tested the technique by measuring increases in dihydroxybenzoate levels after exposure of platelets to experimentally induced oxidative stress. Then, we measured platelet levels of 2,3- and 2,5-DHB in patients with Parkinson's disease, under therapy with L-DOPA, and in normal subjects. We also measured platelet concentrations of L-DOPA and its major metabolite, 3-O-methyldopa (3-OMD). Parkinsonian patients showed increased levels of both 2,3- and 2,5-DHB. Platelet levels of 2,3-DHB were positively correlated with platelet levels of L-DOPA and 3-OMD. The technique we describe proved simple and extremely sensitive and may represent a useful tool for the study of oxidative stress in humans. Copyright (C) 1999.
| Original language | English |
|---|---|
| Pages (from-to) | 213-220 |
| Number of pages | 8 |
| Journal | Journal of Chromatography B: Biomedical Sciences and Applications |
| Volume | 732 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Sep 10 1999 |
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Keywords
- Dihydroxybenzoate
- Hydroxyl free radical formation
- L-DOPA
ASJC Scopus subject areas
- Chemistry(all)
Cite this
Determination of hydroxyl free radical formation in human platelets using high-performance liquid chromatography with electrochemical detection. / Blandini, F.; Martignoni, E.; Ricotti, R.; Di Jeso, F.; Nappi, G.
In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 732, No. 1, 10.09.1999, p. 213-220.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Determination of hydroxyl free radical formation in human platelets using high-performance liquid chromatography with electrochemical detection
AU - Blandini, F.
AU - Martignoni, E.
AU - Ricotti, R.
AU - Di Jeso, F.
AU - Nappi, G.
PY - 1999/9/10
Y1 - 1999/9/10
N2 - The formation of the hydroxyl free radical (HFR) can be quantified indirectly, by measuring two products of the hydroxylation of salicylic acid, 2,3-dihydroxybenzoate (2,3-DHB) and 2,5-dihydroxybenzoate (2,5-DHB). In this study, we used reversed-phase high-performance liquid chromatography with electrochemical (coulometric) detection to measure 2,3- and 2,5-DHB levels in human platelets. The limits of detection of the method were 10 and 5 fmol on column for 2,3-DHB and 2,5-DHB, respectively. We tested the technique by measuring increases in dihydroxybenzoate levels after exposure of platelets to experimentally induced oxidative stress. Then, we measured platelet levels of 2,3- and 2,5-DHB in patients with Parkinson's disease, under therapy with L-DOPA, and in normal subjects. We also measured platelet concentrations of L-DOPA and its major metabolite, 3-O-methyldopa (3-OMD). Parkinsonian patients showed increased levels of both 2,3- and 2,5-DHB. Platelet levels of 2,3-DHB were positively correlated with platelet levels of L-DOPA and 3-OMD. The technique we describe proved simple and extremely sensitive and may represent a useful tool for the study of oxidative stress in humans. Copyright (C) 1999.
AB - The formation of the hydroxyl free radical (HFR) can be quantified indirectly, by measuring two products of the hydroxylation of salicylic acid, 2,3-dihydroxybenzoate (2,3-DHB) and 2,5-dihydroxybenzoate (2,5-DHB). In this study, we used reversed-phase high-performance liquid chromatography with electrochemical (coulometric) detection to measure 2,3- and 2,5-DHB levels in human platelets. The limits of detection of the method were 10 and 5 fmol on column for 2,3-DHB and 2,5-DHB, respectively. We tested the technique by measuring increases in dihydroxybenzoate levels after exposure of platelets to experimentally induced oxidative stress. Then, we measured platelet levels of 2,3- and 2,5-DHB in patients with Parkinson's disease, under therapy with L-DOPA, and in normal subjects. We also measured platelet concentrations of L-DOPA and its major metabolite, 3-O-methyldopa (3-OMD). Parkinsonian patients showed increased levels of both 2,3- and 2,5-DHB. Platelet levels of 2,3-DHB were positively correlated with platelet levels of L-DOPA and 3-OMD. The technique we describe proved simple and extremely sensitive and may represent a useful tool for the study of oxidative stress in humans. Copyright (C) 1999.
KW - Dihydroxybenzoate
KW - Hydroxyl free radical formation
KW - L-DOPA
UR - http://www.scopus.com/inward/record.url?scp=0032852976&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032852976&partnerID=8YFLogxK
U2 - 10.1016/S0378-4347(99)00286-8
DO - 10.1016/S0378-4347(99)00286-8
M3 - Article
C2 - 10517238
AN - SCOPUS:0032852976
VL - 732
SP - 213
EP - 220
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
SN - 1387-2273
IS - 1
ER -