title: Intra-/inter-laboratory validation study on reactive oxygen species assay for chemical photosafety evaluation using two different solar simulators.
abstract: A previous multi-center validation study demonstrated high transferability and reliability of reactive oxygen species (ROS) assay for photosafety evaluation. The present validation study was undertaken to verify further the applicability of different solar simulators and assay performance. In 7 participating laboratories, 2 standards and 42 coded chemicals, including 23 phototoxins and 19 non-phototoxic drugs/chemicals, were assessed by the ROS assay using two different solar simulators (Atlas Suntest CPS series, 3 labs; and Seric SXL-2500V2, 4 labs). Irradiation conditions could be optimized using quinine and sulisobenzone as positive and negative standards to offer consistent assay outcomes. In both solar simulators, the intra- and inter-day precisions (coefficient of variation; CV) for quinine were found to be below 10%. The inter-laboratory CV for quinine averaged 15.4% (Atlas Suntest CPS) and 13.2% (Seric SXL-2500V2) for singlet oxygen and 17.0% (Atlas Suntest CPS) and 7.1% (Seric SXL-2500V2) for superoxide, suggesting high inter-laboratory reproducibility even though different solar simulators were employed for the ROS assay. In the ROS assay on 42 coded chemicals, some chemicals (ca. 19-29%) were unevaluable because of limited solubility and spectral interference. Although several false positives appeared with positive predictivity of ca. 76-92% (Atlas Suntest CPS) and ca. 75-84% (Seric SXL-2500V2), there were no false negative predictions in both solar simulators. A multi-center validation study on the ROS assay demonstrated satisfactory transferability, accuracy, precision, and predictivity, as well as the availability of other solar simulators.
authors.affiliation: Department of Pharmacokinetics and Pharmacodynamics, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan. Electronic address: onoue@u-shizuoka-ken.ac.jp.
authors.forename: Satomi
authors.initials: S
authors.lastname: Onoue
authors.affiliation: Non-Clinical Research Group, Ophthalmic Research and Development Center, Santen Pharmaceutical Co., Ltd., 8916-16 Takayama-cho, Ikoma, Nara 630-0101, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Hosoi
authors.affiliation: Drug Developmental Research Laboratories, Shionogi & Co., Ltd., 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan.
authors.forename: Tsuguto
authors.initials: T
authors.lastname: Toda
authors.affiliation: Research Center, Taisho Pharmaceutical Co., Ltd., 1-403, Yoshino-cho, Kita-ku, Saitama 331-9530, Japan.
authors.forename: Hironori
authors.initials: H
authors.lastname: Takagi
authors.affiliation: Research Center, Taisho Pharmaceutical Co., Ltd., 1-403, Yoshino-cho, Kita-ku, Saitama 331-9530, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Osaki
authors.affiliation: Safety Research Department, ASKA Pharmaceutical Co., Ltd., 5-36-1, Shimosakunobe, Takatsu-ku, Kawasaki, Kanagawa 213-8522, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Matsumoto
authors.affiliation: Pharmaceuticals Research Center, Asahi Kasei Pharma Corporation, 632-1 Mifuku Izunokuni-shi, Shizuoka 410-2321, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Kawakami
authors.affiliation: Laboratory of Cell Toxicology, Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano, Kanagawa 257-8523, Japan.
authors.forename: Shinobu
authors.initials: S
authors.lastname: Wakuri
authors.affiliation: Safety Research Laboratories, Mitsubishi Tanabe Pharma Corporation, 1-1-1, Kazusakamatari, Kisarazu, Chiba 292-0818, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Iwase
authors.affiliation: Safety Research Laboratories, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawaguchi, Toda-shi, Saitama 335-8505, Japan.
authors.forename: Toshinobu
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: Product Development Regulatory Affairs Department, Shionogi & Co., Ltd., 2-17-5 Shibuya, Shibuya-ku, Tokyo 150-8673, Japan.
authors.forename: Kazuichi
authors.initials: K
authors.lastname: Nakamura
authors.affiliation: National Institute of Health Sciences (NIHS), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Ohno
authors.affiliation: Japanese Center for the Validation of Alternative Methods (JaCVAM), National Institute of Health Sciences (NIHS), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Kojima
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001681:Biological Assay; D007753:Laboratories / Q000592:standards*; D051379:Mice; D010777:Photochemistry / Q000379:methods; D017319:Photosensitizing Agents / Q000737:chemistry / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015203:Reproducibility of Results; D013472:Sunlight; D014466:Ultraviolet Rays
chemical_list: D017319:Photosensitizing Agents; D017382:Reactive Oxygen Species
keywords: Inter-laboratory precision; Photoreactivity; Phototoxicity; Reactive oxygen species; Validation