Sarcophagid quinone isomerase readily converted NBAD quinone to its quinone methide, which reacted rapidly with water forming the Michael-1,6-addition product, NBANE. Phenoloxidase generated NBANE quinone served as a substrate for quinone isomerase generating N-β-alanylarterenone. HPLC Mass spectral studies revealed the productions of NBANE, N-β-alanylarterenone, dehydro NBAD, and oligomers of dehydro NBAD in reaction mixture containing NBAD, phenoloxidase, quinone isomerase and quinone methide isomerase.

Tyrosinase oxidized dehydro NBAD to a transient quinone methide imine amide (QMIA) before producing oligomers of dehydro NBAD. Liquid chromatography mass spectrometry analysis of the reaction confirmed the production of dimers and other oligomers. Laccase also oxidized dehydro NBAD to oligomers, with out producing QMIA, probably by free radical coupling. Dehydro NBAD is sensitive to alkaline pH and exhibited rapid nonenzymatic oxidation. These results indicate that dehydro NBAD behaves much like dehydro NADA. The difference in color of cuticle using NBAD and NADA seems to come from the differences in reactivity of NBAD quinone, NBAD quinone methide, and NBAD QMIA and the way the enzymes utilize these reactive intermediates both temporal and spatial in the cuticle during sclerotization.