Amino Reactivity of Glutardialdehyde and Monoaldehydes─Chemoassay Profile vs Skin Sensitization Potency

Chemoassay profiling of organic electrophiles through the direct peptide reactivity assay has become an OECD-accepted nonanimal component in the REACH evaluation of potential skin sensitizers. For aldehydes forming imines (Schiff bases), however, existing chemoassays yielded inconclusive results, indicating issues with their NH₂ sensitivity and the reversibility of the reaction. In the present study, a new kinetic chemoassay employing the N terminus of glycine-para-nitroanilide, Gly-pNA, as a model nucleophile for protein NH₂ groups is introduced and applied to nine aliphatic monoaldehydes and glutardialdehyde (1,5-pentanedial) that have log K_ow (octanol/water partition coefficient) values from 0.63 to 3.99. The Gly-pNA second-order rate constants k₁ range from 8.56 to 150 L·mol^-1·min^-1 for the monoaldehydes. Interestingly, glutardialdehyde with a k₁ of 17 731 L·mol^-1·min^-1 is 170-fold more reactive than its monoaldehyde counterpart pentanal. This can be rationalized by hydration or tautomerization of the dialdehyde to monoaldehydic forms, now facilitating Schiff base formation through an intramolecular H bond. Comparison with murine local lymph node assay data from the literature reveals that adduct stability in terms of reaction thermodynamics (K = k₁/k_-1^pseudo) rather than formation kinetics (k₁) governs the skin sensitization potency of Schiff-base-forming aldehydes. The discussion includes analytically determined adduct patterns, and the impact of α- and β-carbon substitution as well as hydrophobicity on aldehyde reactivity.