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9420087 
Journal Article 
Development and performance evaluation of a moderate positive reference material containing genapol x-080 for hemolysis testing 
Nomura, Y; Shindo, T; Motohashi, H; Yamakage, K; Watanabe, M; Fukui, C; Morishita, Y; Haishima, Y 
2019 
0 
Kokuritsu Iyakuhin Shokuhin Eisei Kenkyūjo Hōkoku / Bulletin of the National Institute of Health Sciences
ISSN: 1343-4292 
National Institute of Health Sciences 
2019 
137 
32-39 
English 
In the previously study, we developed plasticized polyvinyl chloride (PVC) pellets named“Y-2”or “Y-4”that show weak or strong hemolytic activity depending on the content of Genapol X-080, which is used in biological safety evaluations of medical devices. However, the hemolytic activity of Y-2 was weak and inferior in reproducibility tests. This study deals with the development and performance evaluation of a moderate positive reference material (Y-3) for hemolysis testing. Y-3 was prepared as PVC pellets consisting of 33.4% di (2-ethylhexyl)-phthalate (DEHP), 4.86% epoxidized soybean oil (ESBO), 3.04 × 10−2% calcium and zinc stearates, and 0.91% Genapol X-080 as the hemolytic substance. Y-3 showed a hemolysis ratio around 40‒60% with human and rabbit blood samples in American Society for Testing and Materials (ASTM) direct contact assays under the test sample/ extraction vehicle ratio specified by ISO 10993-12 (0.2 g/mL). Y-3 also exhibited hemolytic activity in extract-based assays, but the hemolysis ratio varied greatly depending on the test conditions; this likely occurred because the solubility of Genapol X-080 decreases markedly at temperatures higher than the cloud point (74‒76℃). As a result of optimizing the test sample/extraction vehicle ratio for each extraction condition used in the extract-based assays, a hemolysis ratio around 20‒80% was yielded with human and rabbit blood when Y-3 extracts with phosphate-buffered saline were prepared under the following conditions: 0.08 g/mL at 37 or 50℃ for 72 h, 0.24 g/mL at 70℃ for 24 h, and 0.8 g/ mL at 121℃ for 1 h. Y-3 may be useful as a positive reference material for hemolysis testing and may show advantages over PVC sheets spiked with 0.61% (Y-2: weak positive reference material) or 5.8% (Y-4: strong positive reference material) of Genapol X-080 that induce weak or complete hemolysis, respectively, as previously reported. Hatano Research Institute is planning to start distributing Y-3 as a first positive reference material for hemolysis test on a worldwide scale beginning in the spring of 2019. © 2019, National Institute of Health Sciences. All rights reserved. 
Biological safety evaluation; Guidance; Hemolysis test; Positive reference material