Disclosed are new acridone dye derivatives having characteristic
fluorescence lifetimes. Also disclosed are methods for labelling target
biological materials employing the acridone dyes and use of the labelled
materials in biological assays. The acridone derivatives have the
following structure:
in which Z.sup.1 and Z.sup.2 represent the atoms necessary to complete one
ring, two fused ring, or three fused ring aromatic or heteroaromatic
systems, each ring having five or six atoms selected from carbon atoms
and optionally no more than two atoms selected from oxygen, nitrogen and
sulphur; R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are selected from
hydrogen, halogen, amide, hydroxyl, cyano, nitro, mono- or
di-nitro-substituted benzyl, amino, mono- or di-C.sub.1-C.sub.4
alkyl-substituted amino, sulphydryl, carbonyl, carboxyl, C.sub.1-C.sub.6
alkoxy, acrylate, vinyl, styryl, aryl, heteroaryl, C.sub.1-C.sub.20
alkyl, aralkyl, sulphonate, sulphonic acid, quaternary ammonium, the
group -E-F and the group --(CH.sub.2--).sub.nY; R.sup.1 is selected from
hydrogen, mono- or di-nitro-substituted benzyl, C.sub.1-C.sub.20 alkyl,
aralkyl, the group -E-F and the group --(CH.sub.2--).sub.nY; where E is a
spacer group, F is a target bonding group; Y is selected from sulphonate,
sulphate, phosphonate, phosphate, quaternary ammonium and carboxyl; and n
is an integer from 1 to 6. The invention also relates to a set of
different fluorescent acridone dye derivatives, each dye having a
different fluorescence lifetime, the set of dyes being particularly
useful for multiparameter analysis.