Fully automatic flow method for the determination of scavenging capacity against nitric oxide radicals

• Type: Article in International Scientific Journal
• Date: 2010

Authors

Joana P N Ribeiro (Author) Other; Luis M Magalhaes (Author) Other; Marcela A Segundo (Author) FFUP; Salette Reis (Author) FFUP; Jose L F C Lima (Author) FFUP

Journal

Title: Analytical and Bioanalytical Chemistry

Vol. 397 No. 7

Pages: 3005-3014

ISSN: 1618-2642

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 6 Citations

ISI Web of Science

Scopus - 7 Citations

Scientific classification

CORDIS: Health sciences

FOS: Natural sciences > Chemical sciences

Other information

Authenticus ID: P-003-43B

DOI: 10.1007/s00216-010-3883-5

Resumo (PT): 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 <br> <br> <a target="_blank" href="http://web.ebscohost.com/ehost/detail?hid=104&sid=2ede759a-b305-4a81-b284-b4bf38d0dbbf%40sessionmgr111&vid=5&bdata=JnNpdGU9ZWhvc3QtbGl2ZQ%3d%3d#db=a9h&AN=52299525"> Texto integral </a> <br> <br>

Abstract (EN): 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 degrees C), pH (7.4), and concentration of NO (less than 1 mu M). This approach allowed the evaluation of scavenging capacity against NO by endogenous antioxidant molecules, pharmaceutical compounds, and human plasma. IC(50) values were calculated for rutin (1.30+/-0.02 mu M, positive control), cysteine (321+/-8 mu M), reduced glutathione (1106+/-93 mu M), uric acid (134+/-12 mu M), dipyrone (1.36+/-0.06 mu M), and captopril (363+/-28 mu M). A high degree of automation was attained as the successive dilution of antioxidant standard solutions required for IC(50) 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 10