To establish further a practical quantitative in chemico reactivity assay for screening contact
allergens,
lysine peptide was incorporated into a liquid chromatography and tandem mass spectrometry-based assay for reactivity assessments of
hapten and pre-/pro-
hapten chemical sensitizers. Loss of
peptide was determined following 24 h coincubation with test chemical using a concentration-response study design. A total of 70 chemicals were tested in discrete reactions with
cysteine or
lysine peptide, in the presence and absence of
horseradish peroxidase-
hydrogen peroxide oxidation system. An empirically derived prediction model for discriminating sensitizers from nonsensitizers resulted in an accuracy of 83% for 26
haptens, 19 pre-/pro-
haptens, and 25 nonsensitizers. Four sensitizers were shown to selectively react with
lysine including two strong/extreme and two weak sensitizers. In addition, seven sensitizers were identified as having higher reactivity toward
lysine compared with
cysteine. The majority of sensitizing chemicals (27/45) were reactive toward both
cysteine and
lysine peptides. An estimate of the relative reactivity potency was determined based on the concentration of test chemical that depletes
peptide at or above a threshold positive value. Here, we report the use of EC15 as one example to illustrate the use of the model for screening the skin sensitization potential of novel chemicals. Results from this initial assessment highlight the utility of
lysine for assessing a chemical's potential to elicit sensitization reactions or induce
hypersensitivity. This approach has the potential to qualitatively and quantitatively evaluate an important mechanism in contact
allergy for hazard and quantitative risk assessments without animal testing.