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    PROCESS FOR DETERMINING S-NITROSOTHIOLS IN BIOLOGICAL FLUIDS
    1.
    发明公开
    PROCESS FOR DETERMINING S-NITROSOTHIOLS IN BIOLOGICAL FLUIDS 有权
    用于确定S-亚硝基硫醇生物体液

    公开(公告)号：EP2054727A2

    公开(公告)日：2009-05-06

    申请号：EP07789530.8

    申请日：2007-07-23

    申请人： UNIVERSITA' DI PISA

    发明人： POMPELLA, Alfonso ,  PAOLICCHI, Aldo ,  BRAMANTI, Emilia ,  BARSACCHI, Renata ,  FRANZINI, Maria ,  VECOLI, Cecilia

    IPC分类号： G01N33/68

    CPC分类号： C12Q1/37 ,  C12Q1/48 ,  G01N33/6815 ,  G01N33/6842 ,  G01N33/94 ,  G01N2333/9108 ,  G01N2333/96425 ,  G01N2400/00

    摘要： A process for determining S-nitrosothiols, in particular S-nitrosoglutathione, in biological fluids that is easy, selective, cheap with respect to the prior art, which requires the use of equipment commonly available in laboratories, at low cost, which can be used by not qualified operators. The process is based on the hydrolysis of S-nitrosoglutathione (GSNO) by an enzyme, in particular γ-glutamyltranspeptidase (GGT). This enzyme hydrolizes the residual γ-glutamyl of GSNO for giving glutamate (GIu) and S-nitroso-cysteinylglycine (GIyCySNO). In the presence of ions of transition metals GGT speeds up the release of NO since the intermediate that is formed, the GIyCySNO, is is much more sensitive to a metal- dependent decomposition. Advantageously, the amount of nitric oxide present in the sample is measured through a reaction thereof with 4,5 diaminof luorescein (DAF-2), said reaction creating a fluorescent compound in an amount proportional to the S-nitrosothiol amount present in the sample. Alternatively, the amount of released NO can be measured by a chemiluminescence analyser, commercially available. In the presence of biological fluids having complex matrix, the introduction of the enzyme is done after separation of the S-nitrosothiol from the other components of the fluid.