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    A simple and sensitive assay for the nitric oxide metabolites nitrite and nitrate in serum, bronchoalveolar lavage and culture media

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    Authors
    van Amsterdam JGC
    Piet JJ
    Zuidema J
    Bisschop A
    Wemer J
    Type
    Report
    Language
    en
    
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    Title
    A simple and sensitive assay for the nitric oxide metabolites nitrite and nitrate in serum, bronchoalveolar lavage and culture media
    Translated Title
    Een eenvoudige en gevoelige bepalingsmethode voor de stikstofoxide-metabolieten nitriet en nitraat in serum, bronchoalveolaire lavagevloeistof en cultuurmedia
    Publiekssamenvatting
    NO (Stikstof mono-oxide) wordt door het NO-synthase uit L-arginine vrijgezet en vertegenwoordigt een belangrijke intra- en intercellulaire boodschapperfunctie. Binnen biologische systemen wordt het NO snel naar nitriet en nitraat geoxideerd. Het totale nitriet is te beschouwen als parameter voor NO-synthase activiteit en wordt gekwantificeerd met behulp van de lichtabsorbtie van het Griess-chromofoor bij 420 nm na reductie van nitraat naar nitriet met Klebsiella pneumoniae (BBD 900.990). Met deze methode kunnen secundaire NO-synthase producten op reproduceerbare wijze in sera en andere eiwitbevattende media, zoals bronchoalveolaire wasvloeistoffen en gekleurde kweekmedia met een detectielimiet van 0,3 muM bepaald worden.<br>
    NO (nitric mono-oxide) released from L-arginine by NO-synthase represents an important intra- and intercellular messenger. Within biological systems NO is rapidly oxidized to nitrite and nitrate. Total nitrite regarded as parameter for NO-synthase activity is quantified using absorbance of the Griess chromophore at 420 nm after deproteination and reduction of nitrate to nitrite using Klebsiella pneumoniae (BBD 900.990). This method allows reproducible assay of secundary NO-synthase products in sera and other protein containing media like bronchoalveolar lavages and coloured culture media with a detection limit of 0.3 muM.<br>
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    Rijksinstituut voor Volksgezondheid en Milieu RIVM
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