Method of determination of a fluorescent substance, and a method of assay of an enzyme activity

摘要

A method for determining a fluorescent substance in a broader concentration range with higher measurement precision is provided which is applicable to assay of enzyme activity in enzyme-label immunoassay, enzyme-label DNA hybridization, etc. A method for assaying enzyme activity is also provided which employs the above fluorescence measurement. In the method, a specimen containing an objective fluorescent substance to be determined and a reference substance is irradiated with excitation light capable of exciting both the objective fluorescent substance and the reference fluorescent substance; measuring a first fluorescence intensity at a first wavelength where fluorescence is emitted mainly by the objective fluorescent substance; measuring a second fluorescence intensity at a second wavelength where fluorescence is emitted mainly by the reference fluorescent substance; and calculating the concentration of the objective fluorescent substance from the equation: $where\; x\; is\; the\; concentration\; of\; the\; objective\; fluorescent\; substance,\; and\; y\; is\; a\; ratio\; of\; the\; first\; fluorescence\; intensity\; to\; the\; second\; fluorescence\; intensity.\mathop{\mathrm{image}}$