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- Title
Fully automatic flow method for the determination of scavenging capacity against nitric oxide radicals.
- Authors
Ribeiro, Joana P. N.; Magalhães, Luís M.; Segundo, Marcela A.; Reis, Salette; Lima, José L. F. C.
- Abstract
In the present work, a fluorimetric automatic method based on multisyringe flow injection analysis (MSFIA) was developed for in vitro evaluation of scavenging capacity against nitric oxide (NO) using 4,5-diaminofluorescein (DAF-2) as an NO-selective fluorogenic probe. The MSFIA manifold was assembled to perform the in-line generation of NO and the competitive reaction of putative scavenger molecules and DAF-2 with NO at conditions close to those found in vivo regarding temperature (37°C), pH (7.4), and concentration of NO (less than 1 μM). This approach allowed the evaluation of scavenging capacity against NO by endogenous antioxidant molecules, pharmaceutical compounds, and human plasma. IC50 values were calculated for rutin (1.30 ± 0.02 μM, positive control), cysteine (321 ± 8 μM), reduced glutathione (1106 ± 93 μM), uric acid (134 ± 12 μM), dipyrone (1.36 ± 0.06 μM), and captopril (363 ± 28 μM). A high degree of automation was attained as the successive dilution of antioxidant standard solutions required for IC50 assessment was performed automatically, in a dilution chamber placed in the flow system. A determination throughput of 16 h-1 and a good precision were attained (relative standard deviation between 1.6 and 9.0%), fostering the application of the proposed method to routine/screening analysis of scavenging capacity against NO. [Figure not available: see fulltext.]
- Subjects
RADICALS (Chemistry); NITRIC oxide; FLOW injection analysis; FLUORIMETRY; PRECIPITATION scavenging
- Publication
Analytical & Bioanalytical Chemistry, 2010, Vol 397, Issue 7, p3005
- ISSN
1618-2642
- Publication type
Article
- DOI
10.1007/s00216-010-3883-5