Establishment and intra-/inter-laboratory validation of a standard protocol of reactive oxygen species assay for chemical photosafety evaluation.
A reactive oxygen species (ROS) assay was previously developed for photosafety evaluation of pharmaceuticals, and the present multi-center study aimed to establish and validate a standard protocol for ROS assay. In three participating laboratories, two standards and 42 coded chemicals, including 23 phototoxins and 19 nonphototoxic drugs/chemicals, were assessed by the ROS assay according to the standardized protocol. Most phototoxins tended to generate singlet oxygen and/or superoxide under UV-vis exposure, but nonphototoxic chemicals were less photoreactive. In the ROS assay on quinine (200 µm), a typical phototoxic drug, the intra- and inter-day precisions (coefficient of variation; CV) were found to be 1.5-7.4% and 1.7-9.3%, respectively. The inter-laboratory CV for quinine averaged 15.4% for singlet oxygen and 17.0% for superoxide. The ROS assay on 42 coded chemicals (200 µm) provided no false negative predictions upon previously defined criteria as compared with the in vitro/in vivo phototoxicity, although several false positives appeared. Outcomes from the validation study were indicative of satisfactory transferability, intra- and inter-laboratory variability, and predictive capacity of the ROS assay.
Department of Pharmacokinetics and Pharmacodynamics, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan.
Satomi
S
Onoue
Kazuhiro
K
Hosoi
Shinobu
S
Wakuri
Yumiko
Y
Iwase
Toshinobu
T
Yamamoto
Naoko
N
Matsuoka
Kazuichi
K
Nakamura
Tsuguto
T
Toda
Hironori
H
Takagi
Naoto
N
Osaki
Yasuhiro
Y
Matsumoto
Satoru
S
Kawakami
Yoshiki
Y
Seto
Masashi
M
Kato
Shizuo
S
Yamada
Yasuo
Y
Ohno
Hajime
H
Kojima
D001577:Benzophenones / Q000737:chemistry / Q000528:radiation effects; D017484:Dermatitis, Phototoxic / Q000209:etiology / Q000378:metabolism; D064420:Drug-Related Side Effects and Adverse Reactions / Q000378:metabolism; D006801:Humans; D007753:Laboratories / Q000592:standards*; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000528:radiation effects*; D055668:Photochemical Processes; D011803:Quinine / Q000737:chemistry / Q000528:radiation effects; D017382:Reactive Oxygen Species / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results; D014466:Ultraviolet Rays / Q000009:adverse effects; D054928:Validation Studies as Topic
D001577:Benzophenones; D004364:Pharmaceutical Preparations; D017382:Reactive Oxygen Species; C009361:sulisobenzone; D011803:Quinine