Within‐ and between‐laboratory reproducibility and predictive capacity of amino acid derivative reactivity assay (ADRA) using a 0.5?mg/mL test chemical solution: Results of the study for reproducibility confirmation implemented in five participating laboratories

Yusuke Yamamoto; Masaharu Fujita; Shinichi WatanabeHiroaki YamagaKoji WakabayashiYu TaharaNobuyuki HorieKeiichi FujimotoKazuya TakeuchiKohei KamiyaTsuyoshi KawakamiKohichi KojimaTakashi SozuHajime KojimaToshihiko KasaharaAtsushi Ono

首页> 外文期刊>Journal of applied toxicology >Within‐ and between‐laboratory reproducibility and predictive capacity of amino acid derivative reactivity assay (ADRA) using a 0.5?mg/mL test chemical solution: Results of the study for reproducibility confirmation implemented in five participating laboratories

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Within‐ and between‐laboratory reproducibility and predictive capacity of amino acid derivative reactivity assay (ADRA) using a 0.5?mg/mL test chemical solution: Results of the study for reproducibility confirmation implemented in five participating laboratories

机译：Within‐ and between‐laboratory reproducibility and predictive capacity of amino acid derivative reactivity assay (ADRA) using a 0.5?mg/mL test chemical solution: Results of the study for reproducibility confirmation implemented in five participating laboratories

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Abstract The amino acid derivative reactivity assay (ADRA) is an in chemico alternative assay for skin sensitization listed in OECD test guideline 442C. ADRA evaluates the reactivity of sensitizers to proteins, which is key event 1 in the skin sensitization adverse outcome pathway. Although the current key event 1 evaluation method is a simple assay that evaluates nucleophile and test chemical reactivity, mixtures of unknown molecular weights cannot be evaluated because a constant molar ratio between the nucleophile and test chemical is necessary. In addition, because the nucleophile is quantified by HPLC, the frequency of co‐eluting the test chemical and nucleophile increases when measuring multi‐component mixtures. To solve these issues, test conditions have been developed using a 0.5?mg/mL test chemical solution and fluorescence‐based detection. Since the practicality of these methods has not been substantiated, a validation test to confirm reproducibility was conducted in this study. The 10 proficiency substances listed in the ADRA guidelines were tested three times at five different laboratories. The results of both within‐ and between‐laboratory reproducibility were 100, and the results of ultraviolet‐ and fluorescence‐based measurements were also consistent. In addition to the proficiency substances, a new positive control, squaric acid diethyl ester, was tested three times at the five laboratories. The results showed high reproducibility with N‐(2‐(1‐naphthyl)acetyl)‐l‐cysteine depletion of 37–52 and α‐N‐(2‐(1‐naphthyl)acetyl)‐l‐lysine depletion of 99–100. Thus, high reproducibility was confirmed in both evaluations of the 0.5?mg/mL test chemical and the fluorescence‐based measurements, validating the practicability of these methods.

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《Journal of applied toxicology》 |2022年第6期|1078-1090|共13页
作者
Yusuke Yamamoto; Masaharu Fujita; Shinichi WatanabeHiroaki YamagaKoji WakabayashiYu TaharaNobuyuki HorieKeiichi FujimotoKazuya TakeuchiKohei KamiyaTsuyoshi KawakamiKohichi KojimaTakashi SozuHajime KojimaToshihiko KasaharaAtsushi Ono;
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作者单位

Safety Evaluation Center,Fujifilm Corporation;

Safety Science Research Laboratories,Lion Corporation;

Chemical Safety Department,Mitsui Chemicals, Inc.Environmental Health Science Laboratory,Sumitomo Chemical Co., Ltd.Biological Research Laboratories,Nissan Chemical CorporationDivision of Environmental Chemistry,National Institute of Health SciencesFood and Drug Safety CenterFaculty of Engineering,Tokyo University of ScienceBiological safety Research Center, Division of Risk Assessment, National Institute of HealthGraduate School of Medicine, Dentistry and Pharmaceutical Sciences, Division of Pharmaceutical;

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正文语种英语
中图分类毒物学（毒理学）;
关键词
ADRA (0.5?mg/mL); amino acid derivative reactivity assay; between‐laboratory reproducibility; NAC; NAL; predictive capacity; proficiency substances; reproducibility; within‐laboratory reproducibility;

Within‐ and between‐laboratory reproducibility and predictive capacity of amino acid derivative reactivity assay (ADRA) using a 0.5?mg/mL test chemical solution: Results of the study for reproducibility confirmation implemented in five participating laboratories

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