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QUANTIFICATION OF NITRITE IN THE PRESENCE OF ANTHOCYAMNS USING GRIESS AND GC/MS ASSAYS 1
Author(s) -
WANG J.,
MAZZA G.
Publication year - 2002
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.2002.tb00758.x
Subject(s) - griess test , nitrite , chemistry , chromatography , nitric oxide , absorbance , repeatability , reagent , lipopolysaccharide , gas chromatography–mass spectrometry , mass spectrometry , biochemistry , nitrate , organic chemistry , medicine , endocrinology
Nitrite level in cell supernatants is cm index of nitric oxide (NO) produced by lipopolysaccharide/interferon‐γ (LPS/IFN‐y) activated RAW 264.7 macrophages. The Griess assay has been widely used to quantify nitrite in biological fluids including cell supernatants. The assay is based on the reaction of nitrite under acidic conditions with Griess reagent to form a purple azo dye with a maximum wavelength (λ max ) at 540 nm. In acidic media anthocyanins also have intense absorbance at the same visible absorption range as the azo dye. A modified Griess assay is described to quantify nitrite in cell supernatants in the presence of anthocyanin pigments. Gas chromatography mass spectrometry (GC/MS) with electron impact ioniziation (EI) was also used to quantify nitrite but suffered from poor repeatability. Results obtained by the modified Griess and the GC/MS assay were significantly correlated and served to demonstrate the effects of anthocyanins on NO production in RAW 264.7 macrophages .