Abstract: Fluorogenic substrates, methods of making the fluorogenic substrates, and methods of detecting glycosidase activities (EC3.2.1) are provided. The fluorogenic substrates are composed of a sugar covalently bonded to the 7-hydroxy position of a substituted coumarin sulfonated fluorophore is provided. The fluorogenic substrates include sodium ?-D-cellobioside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; sodium ?-D-glucoside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; sodium ?-D-xyloside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; sodium ?-D-xylobioside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; and sodium ?-D-xylopolyoside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate. The fluorogenic substrates may be made by covalently bonding the sugar moiety to the 7-hydroxy position of the substituted coumarin sulfonated fluorophore by reaction in DMF.

Abstract: Substituted sulfonated coumarins are expressed in the general formula (I), where: R1 is H, OH, or a substituted or unsubstituted, straight or branched C1-C6 alkyl radical, or —COR4, or —COOR4, or —CONHR4, R2 is H or a halogen, in particular fluorine, or a substituted or unsubstituted, straight or branched C1-C6 alkyl radical, or —COR4, or —COOR4, or —CONHR4, R1 and R2 being capable of together forming a ring, such as a substituted or unsubstituted aryl or furane, R3 is H or a halogen, in particular fluorine, or a substituted or unsubstituted, straight or branched C1-C6 alkyl radical, or —COR4, or —COOR4, or —CONHR4, where R4 is H, or a substituted or unsubstituted, straight or branched C1-C6 alkyl radical, or a substituted or unsubstituted aryl, and M is Na or K.