4-Nitrophthalic acid and 1,3-benzenediol were fused 6 h/190-200degreesC (no dehydrant); the product was dissolved in dild. NaOH and precipitated with HCl to yield (80%) 5- and 6-nitrofluoresceins. To separate the isomers, the resulting mixture was acetylated (with AcH) and the O-diacetyl derivatives were crystallized from EtOH-toluene. The acetates were hydrolyzed, each nitro was reduced with aq Na2S + NaHS to the amino-fluorescein and purified by crystallization to yield 72% of 5-amino and 60% of 6-aminofluorescein.