Assay of humain platelet guanylate cyclase activity by high-perofrmance liquid chromatography with fluorescence derivatization

摘要

A selective and sensitive high-performance liquid chromatography(HPLC) method with fluorescence derivatization for the assay of guanylate cyclase (GC)activity is described.GTP and cGMP ,which are the substrate and the product of GC,respectively,and other guanine-containing compunds are selectively converted by the reaction with (3,4-dimethoxyphenyl)glyooxal to the fluorescent derivatives.The derivatives were separated by reversed-phase HPLC.The limit of detection at a singnal-to-noise ratio of 3 for cGMP was 10 fmol on the column.The sensitivity of this method was less than that of the conventional radiosotopic metod,but this method is simple and convenient.Human platelet GC activity was measured,and theeffects of some compunds were investigated.