Reversed‐phase high‐performance liquid chromatography method with fluorescence detection to screen nitric oxide synthases inhibitors. Issue 12 (29th April 2014)

• Record Type: Journal Article
• Authors: Maccallini, Cristina; Di Matteo, Mauro; Ammazzalorso, Alessandra; D'Angelo, Alessandra; De Filippis, Barbara; Di Silvestre, Sara; Fantacuzzi, Marialuigia; Giampietro, Letizia; Pandolfi, Assunta; Amoroso, Rosa
• Is Part Of: Journal of separation science. Volume 37:Issue 12(2014:Jun.)
• Page Start: 1380
• Page End: 1385
• Publication Date: 2014-04-29
• Subjects: Separation (Technology) -- Periodicals; Chromatographic analysis -- Periodicals; 543.089
• DOI: 10.1002/jssc.201400059
• Languages: English
• ISSNs: 1615-9306

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Nitric oxide synthase (NOS) inhibitors are potential drug candidates due to the critical role of an excessive production of nitric oxide in a range of diseases. At present, the radiometric detection of <sc>l</sc>‐[³H]‐citrulline produced from <sc>l</sc> ‐[³H]‐arginine during the enzymatic reaction is one of the most accepted methods to assess the <italic>in vitro</italic> activity of NOS inhibitors. Here we report a fast, easy, and cheap reversed‐phase high‐performance liquid chromatography method with fluorescence detection, based on the precolumn derivatization of <sc>l</sc>‐citrulline with <italic>o</italic>‐phthaldialdehyde/<italic>N</italic>‐acetyl cysteine, for the <italic>in vitro</italic> screening of NOS inhibitors. To evaluate enzyme inhibition by the developed method, <italic>N</italic>‐[3‐(aminomethyl)benzyl]acetamidine, a potent and selective inhibitor of inducible NOS, was used as a test compound. The half maximal inhibitory concentration obtained was comparable to that derived by the well‐established radiometric assay.</p> </abstract>