The chemiluminescent (CL) properties of biacridine probes, such as 10,10'-dimethyl-3,3'-disulfo-9,9'-biacridine (DMDSBA), 3,3'-disulfo-9,9'-biacridine dinitrate (DSBADN), 10,10'-diethcarboxyl-9,9'-biacridine dinitrate (DEBADN), 10,10'-dimethyl-3,3'-diamino-9,9'-biacridine (DMDABA), and 10,10'-di(4-aminobutyl)-9,9-biacridine dinitrate (DABADN), were studied in this paper, and the relationships between properties and molecular structures were obtained. A new theory and some valuable experimental data have been provided for the investigation and exploitation of the new CL probes for biomacromolecules.