Abstract
We developed a double microdialysis fiber technique to generate hydroxyl radicals (OH) in rat spinal cord. H2O2 and FeCl2/EDTA were pumped through two parallel microdialysis fibers inserted into the spinal cord such that the reactants mix in the tissue to generate OH by the Fenton reaction. Generated OH was detected by administering phenylalanine through one fiber and measuring o-,m- and p-hydroxyphenylalanine in collected, dialysates by high pressure liquid chromatography and fluorescence detection. The hydroxyphenylalanines are produced by OH attacking the phenylalanine. OH generation was also accomplished in in vitro experiments and the results were consistent with in vivo experiments. This novel method to generate and measure OH radical in vivo overcomes difficulties in studying damage to tissue by short-lived OH. Although developed to study the role of OH in spinal cord injury, this method could be used to study other diseases involving OH damage.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 281-291 |
| Number of pages | 11 |
| Journal | Journal of Biochemical and Biophysical Methods |
| Volume | 27 |
| Issue number | 4 |
| DOIs | |
| State | Published - 1993 |
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Keywords
- Fluorescence detection
- High pressure liquid chromatography
- Hydroxyl radical generation
- Microdialysisl Fenton reaction
- Phenylalanine hydroxylation
ASJC Scopus subject areas
- Biochemistry
- Biophysics
Cite this
Generation and detection of hydroxyl radical in vivo in rat spinal cord by microdialysis administration sampling. / Liu, Danxia.
In: Journal of Biochemical and Biophysical Methods, Vol. 27, No. 4, 1993, p. 281-291.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Generation and detection of hydroxyl radical in vivo in rat spinal cord by microdialysis administration sampling
AU - Liu, Danxia
PY - 1993
Y1 - 1993
N2 - We developed a double microdialysis fiber technique to generate hydroxyl radicals (OH) in rat spinal cord. H2O2 and FeCl2/EDTA were pumped through two parallel microdialysis fibers inserted into the spinal cord such that the reactants mix in the tissue to generate OH by the Fenton reaction. Generated OH was detected by administering phenylalanine through one fiber and measuring o-,m- and p-hydroxyphenylalanine in collected, dialysates by high pressure liquid chromatography and fluorescence detection. The hydroxyphenylalanines are produced by OH attacking the phenylalanine. OH generation was also accomplished in in vitro experiments and the results were consistent with in vivo experiments. This novel method to generate and measure OH radical in vivo overcomes difficulties in studying damage to tissue by short-lived OH. Although developed to study the role of OH in spinal cord injury, this method could be used to study other diseases involving OH damage.
AB - We developed a double microdialysis fiber technique to generate hydroxyl radicals (OH) in rat spinal cord. H2O2 and FeCl2/EDTA were pumped through two parallel microdialysis fibers inserted into the spinal cord such that the reactants mix in the tissue to generate OH by the Fenton reaction. Generated OH was detected by administering phenylalanine through one fiber and measuring o-,m- and p-hydroxyphenylalanine in collected, dialysates by high pressure liquid chromatography and fluorescence detection. The hydroxyphenylalanines are produced by OH attacking the phenylalanine. OH generation was also accomplished in in vitro experiments and the results were consistent with in vivo experiments. This novel method to generate and measure OH radical in vivo overcomes difficulties in studying damage to tissue by short-lived OH. Although developed to study the role of OH in spinal cord injury, this method could be used to study other diseases involving OH damage.
KW - Fluorescence detection
KW - High pressure liquid chromatography
KW - Hydroxyl radical generation
KW - Microdialysisl Fenton reaction
KW - Phenylalanine hydroxylation
UR - http://www.scopus.com/inward/record.url?scp=0027359748&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027359748&partnerID=8YFLogxK
U2 - 10.1016/0165-022X(93)90009-D
DO - 10.1016/0165-022X(93)90009-D
M3 - Article
VL - 27
SP - 281
EP - 291
JO - Journal of Proteomics
JF - Journal of Proteomics
SN - 1874-3919
IS - 4
ER -