TY - JOUR
T1 - Nitrosothiol quantification in human plasma
AU - Goldman, Robert K.
AU - Vlessis, Angelo A.
AU - Trunkey, Donald D.
PY - 1998/5/15
Y1 - 1998/5/15
N2 - A high-pressure liquid chromatography (HPLC) assay for measuring picomole quantities of nitrosothiol in biological samples was developed. The assay utilizes the catalytic reduction of nitrosothiol by mercuric cation (Hg2+). Released nitrogen oxide reacts with sulfanilamide (SA) and N-(1- napthyl)ethylenediamine (NNED) to form a stable azo dye. The azo dye is then separated from N-(1-napthyl)ethylenediamine and quantified by reversed-phase HPLC. In addition to nitrosothiol, nitrite and atmospheric nitrogen oxides are sources of nitrogen oxide that react with the reagents, SA and NNED, to form the azo dye. Therefore, a reference sample, which includes the nitrosothiol sample and all reagents except Hg2+, is utilized for the subtraction of nitrite and atmospheric nitrogen oxides which 'contaminate' the nitrosothiol sample and reagents. This method is a sensitive (~3 pmol; ~10-1 μM) and accurate means to measure nitrosothiol concentration in biologic samples.
AB - A high-pressure liquid chromatography (HPLC) assay for measuring picomole quantities of nitrosothiol in biological samples was developed. The assay utilizes the catalytic reduction of nitrosothiol by mercuric cation (Hg2+). Released nitrogen oxide reacts with sulfanilamide (SA) and N-(1- napthyl)ethylenediamine (NNED) to form a stable azo dye. The azo dye is then separated from N-(1-napthyl)ethylenediamine and quantified by reversed-phase HPLC. In addition to nitrosothiol, nitrite and atmospheric nitrogen oxides are sources of nitrogen oxide that react with the reagents, SA and NNED, to form the azo dye. Therefore, a reference sample, which includes the nitrosothiol sample and all reagents except Hg2+, is utilized for the subtraction of nitrite and atmospheric nitrogen oxides which 'contaminate' the nitrosothiol sample and reagents. This method is a sensitive (~3 pmol; ~10-1 μM) and accurate means to measure nitrosothiol concentration in biologic samples.
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U2 - 10.1006/abio.1998.2651
DO - 10.1006/abio.1998.2651
M3 - Article
C2 - 9606149
AN - SCOPUS:0032524424
SN - 0003-2697
VL - 259
SP - 98
EP - 103
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -