A chemiluminescent substrate of a hydrolytic
enzyme having the following general Formula I is disclosed, as follows:Lumi-M-PFormula Iwhere “Lumi” is a chemiluminescent
moiety capable of producing light (a) by itself, (b) with MP attached and (c) with M attached. Examples of Lumi include chemiluminescent acridinium compounds, benzacridinium compounds,
quinolinium compounds, isoquinolinium compounds, phenanthridinium compounds, and lucigenin compounds, spiroacridan compounds,
luminol compounds and isoluminol compounds. M is a multivalent
heteroatom having at least one
lone pair of electrons selected from
oxygen,
nitrogen and
sulfur, directly attached to the light emitting
moiety of Lumi at one end and to P at the other end. P is a group that can be readily removed by hydrolytic enzymes. An enzymatic reaction utilizing the above compound is the following:where HE is a hydrolytic
enzyme. Lumi-M is a chemiluminescent product having physical and / or chemical properties different from those of Lumi-M-P.