Assay for s-nitrosothiol compounds

G - Physics – 01 – N

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Details

G01N 33/487 (2006.01) G01N 24/10 (2006.01) G01R 33/60 (2006.01)

Patent

CA 2420496

A method for the quantitative measurement of S-nitrosothiols, e.g. S- nitrosoglutathione (RN=57564-91-7), in a biological sample comprises converting the S-nitrosothiols to nitric acid in alkaline solution (pH>10.5), reacting the nitric oxide with a spin trap, e.g. 3,5-dibromo-4-nitrosobenzene sulphonate (DBNBS), or an iron (II) complex of N-methyl-D-glucamine dithiocarbamate (MGD) else diethylcarbamate (DETC), to produce a paramagnetic adduct, and quantifying the paramagnetic adduct using EPR spectrometry. In a second method, a spin trap capable of reacting with the thiyl radical, e.g. 5- (diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide or 5,5-dimethyl-1-pyrroline- N-oxide, is used. Diagnostic methods based on the quantitative measurement of S-nitrosothiols are also described.

La présente invention concerne un procédé permettant de réaliser des mesures quantitatives sur des S-nitrosothiols tels que S-nitrosoglutathione (RN=57564-91-7) dans un échantillon biologique. Ledit procédé comprend la conversion des S-nitrosothiols en acide nitrique dans une solution alcaline (pH>10,5), la réaction de l'acide nitrique avec un agent de piégeage de spin (spin trap) tel que le 3,5-dibromo-4-nitrosobenzène sulphonate (DBNBS) ou un complexe de fer (II) de N-méthyl-D-glucamine dithiocarbamate (MGD) sinon diéthylcarbamate (DETC), afin de produire un produit d'addition paramagnétique, et la quantification du produit d'addition paramagnétique par spectroscopie EPR. Dans un second procédé, un spin trap capable de réagir avec le radical thiyl tel que le 5-(diéthoxyphosphoryl)-5-méthyl-1-pyrroline-N-oxyde ou 5,5-diméthyl-1-pyrroline-N-oxyde, est utilisé. Cette invention concerne également des procédés de diagnostic se basant sur les mesures quantitatives réalisées sur les S-nitrosothiols.

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