“…The high specificity of the present reaction is due to simultaneous requirement of two different functional groups, active methylene and adjoining carbonyl groups, for the formation of fluorophores. Therefore, indole and pyrrole, which can be detected with colorimetric reagents for active methylenes such as sodium nitroprusside (23,24), 1,2naphthoquinone-4-sulfonic acid (25,26) and p-dimethylaminobenzaldehyde (25), are not detectable in the present fluorescent reaction. Moreover, some other typical active methylenes such as creatinine, resorcinol, orcinol, and phloroglucinol which may produce a cyclic n-methylene carbonyl group and show positive response to the colorimetric reagents or some cyclic -methylene carbonyls such as dimedone, camphor, and isoketopinic acid gave no fluorescence, probably because of the steric hindrance around the amethylene carbonyl groups.…”