A PRACTICAL METHOD FOR PREPARING PEROXYNITRITE SOLUTIONS OF LOW IONIC-STRENGTH AND FREE OF HYDROGEN-PEROXIDE

被引:224
作者
PRYOR, WA
CUETO, R
JIN, X
KOPPENOL, WH
NGUSCHWEMLEIN, M
SQUADRITO, GL
UPPU, PL
UPPU, RM
机构
[1] Biodynamics Institute, Louisiana State University, Baton Rouge, LA
基金
美国国家卫生研究院;
关键词
AZIDE; DIMETHYL SULFOXIDE; HYDROGEN PEROXIDE; NITRIC OXIDE; NITRITE; OZONE; PEROXYNITRITE; L-TYROSINE NITRATION; FREE RADICALS;
D O I
10.1016/0891-5849(94)00105-S
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The reaction of ozone (similar to 5% in oxygen) with sodium azide (0.02-0.2 M in water) at pH 12 and 0-4 degrees C is shown to yield concentrated, stable peroxynitrite solutions of up to 80 mM. The product of this reaction is identified based on a broad absorption spectrum with a maximum around 302 nm and by its first-order rate of decomposition (k = 0.40 +/- 0.01 s(-1) at pH 7.05 and 25 degrees C). These peroxynitrite solutions can be obtained essentially free of hydrogen peroxide (detection limit 1 mu M) and only traces of azide (detection limit 0.1 mM). They are low in ionic strength and have a pH of about 12 but without buffering capacity; therefore, they can be adjusted to any pH by addition of buffer. These preparations of peroxynitrite frozen at -20 degrees C show negligible decomposition for about 3 weeks of storage and follow a first-order decomposition with a halflife of about 7 days at refrigerator temperatures (similar to 5 degrees C). These preparations give reactions that are characteristic of peroxynitrite. For example, at pH 7.0, they react with L-tyrosine to give a 7.3 mol % yield of nitrotyrosine(s), and with dimethyl sulfoxide to give a 8.2 mol % yield of formaldehyde, based on starting peroxynitrite concentration.
引用
收藏
页码:75 / 83
页数:9
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