Utility and limitations of a peptide reactivity assay to predict fragrance allergens in vitro.
A key step in the skin sensitization process is the formation of a covalent adduct between the skin sensitizer and endogenous proteins and/or peptides in the skin. A published peptide depletion assay was used to relate the in vitro reactivity of fragrance molecules to LLNA data. Using the classical assay, 22 of 28 tested moderate to strong sensitizers were positive. The prediction of weak sensitizers proved to be more difficult with only 50% of weak sensitizers giving a positive response, but for some compounds this could also be due to false-positive results from the LLNA. LC-MS analysis yielded the expected mass of the peptide adducts in several cases, whereas in other cases putative oxidation reactions led to adducts of unexpected molecular weight. Several moderately sensitizing aldehydes were correctly predicted by the depletion assay, but no adducts were found and the depletion appears to be due to an oxidation of the parent peptide catalyzed by the test compound. Finally, alternative test peptides derived from a physiological reactive protein with enhanced sensitivity for weak Michael acceptors were found, further increasing the sensitivity of the assay.
Natsch
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Givaudan Schweiz AG, Ueberlandstrasse 138, CH-8600 Duebendorf, Switzerland. andreas.natsch@givaudan.com
Gfeller
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H
Rothaupt
M
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Ellis
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G
D000447:Aldehydes / Q000737:chemistry / Q000633:toxicity; D000485:Allergens / Q000737:chemistry / Q000633:toxicity*; D002853:Chromatography, Liquid; D017449:Dermatitis, Allergic Contact / Q000209:etiology; D005544:Forecasting / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010455:Peptides; D010476:Perfume / Q000737:chemistry / Q000633:toxicity*; D023422:Skin Irritancy Tests / Q000379:methods*
D000447:Aldehydes; D000485:Allergens; D010455:Peptides; D010476:Perfume