关键词: 致敏, 皮肤变态反应, 动力学直接多肽反应试验, OECD TG 442C, 小檗碱

Abstract:

This study systematically investigates the applicability of the kinetic direct peptide reactivity assay （kDPRA）, an alternative method for skin sensitization assessment adopted by the Organisation for Economic Cooperation and Development （OECD） in 2022, in domestic laboratories. By examining the influence of key experimental factors such as reagents, the research combines internal laboratory validation with a ring test involving four external laboratories. The results show that different brands of monobromobimane reagent have no significant impact on the assay results, and commercially available versus laboratory-prepared neutral phosphate buffers yield consistent results under identical formulations. Inter-laboratory validation achieves 100% accuracy in the sensitization potency classification of 10 reference substances, fully aligning with OECD TG 442C requirements. For the 10 blind-coded samples, the predictive accuracy across the five participating laboratories ranges between 80% and 100%, meeting the validation acceptance criteria. Furthermore, kDPRA demonstrates reliable performance when applied within a defined approach for evaluating the sensitization potential of the yellow-colored compound berberine. These findings indicate that the kDPRA method exhibits good reproducibility and reliability in Chinese laboratories. It is suitable for domestic implementation and adoption, and can serve as a component of an Adverse Outcome Pathway （AOP） -based integrated testing strategy for skin sensitization assessment. This work provides scientific evidence and practical support for establishing national kDPRA standards, promoting the method, and improving China’s alternative testing system for animal experimentation.

Key words: sensitization, allergic contact dermatitis, kinetic direct peptide reactivity assay, OECD TG442C, berberine