Development of micellar reactive oxygen species assay for photosafety evaluation of poorly water-soluble chemicals

详细信息    查看全文

• 作者：Yoshiki Seto ; Masashi Kato ; Shizuo Yamada ; Satomi Onoue
• 关键词：CV ; coefficient of variation ; DMSO ; dimethyl sulfoxide ; mROS assay ; micellar reactive oxygen species assay ; NaPB ; sodium phosphate buffer ; NBT ; nitroblue tetrazolium ; OECD ; the Organisation for Economic Co-operation and Development ; PABA ; p-aminobenzoic a
• 刊名：Toxicology in Vitro
• 出版年：2013
• 出版时间：September, 2013
• 年：2013
• 卷：27
• 期：6
• 页码：1838-1846
• 全文大小：825 K

文摘

A reactive oxygen species (ROS) assay was previously developed for photosafety assessment; however, the phototoxic potential of some chemicals cannot be evaluated because of their limited aqueous solubility. The present study was undertaken to develop a new micellar ROS (mROS) assay system for poorly water-soluble chemicals using a micellar solution of 0.5 % (v/v) Tween 20 for solubility enhancement. In repeated mROS assay, intra- and inter-day precisions (coefficient of variation) were found to be below 11 % , and the Z¡ä-factors for singlet oxygen and superoxide suggested a large separation band between positive and negative standards. The ROS and mROS assays were applied to 65 phototoxins and 18 non-phototoxic compounds for comparative purposes. Of all 83 chemicals, 25 were unevaluable in the ROS assay due to poor solubility, but only 2 were in the mROS assay. Upon mROS assay on these model chemicals, the individual specificity was 76.5 % , and the positive and negative predictivities were found to be 93.9 % and 86.7 % , respectively. The mROS assay provided 2 false negative predictions, although negative predictivity for the ROS assay was found to be 100 % . Considering the pros and cons of these assays, strategic combined use of the ROS and mROS assays might be efficacious for reliable photosafety assessment with high applicability and predictivity.