Development and performance evaluation of a positive reference material for hemolysis testing

• Published in: Journal of biomedical materials research. Part B, Applied biomaterials Vol. 102; no. 8; pp. 1809 - 1816
• Main Authors: Haishima, Yuji, Hasegawa, Chie, Nomura, Yusuke, Kawakami, Tsuyoshi, Yuba, Toshiyasu, Shindo, Tomoko, Sakaguchi, Keisuke, Tanigawa, Takahiro, Inukai, Kaori, Takenouchi, Mika, Isama, Kazuo, Matsuoka, Atsuko, Niimi, Shingo
• Format: Journal Article
• Language: English
• Published: Hoboken, NJ Blackwell Publishing Ltd 01.11.2014; Wiley-Blackwell; Wiley Subscription Services, Inc
• Subjects: Animals; Assaying; Biological and medical sciences; biological safety evaluation; Biomedical materials; Contact; guidance; Hematologic Tests - methods; Hematologic Tests - standards; Hemolysis; hemolysis test; Materials research; Materials science; Medical sciences; Micelles; Performance evaluation; Polyethylene Glycols - chemistry; Polyvinyl Chloride; Polyvinyl chlorides; positive control; Rabbits; Reference materials; Reference Standards; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Surgical implants; Technology. Biomaterials. Equipments; Performance evaluation; Guidance; Hemolysis; Biological properties; biological safety evaluation; positive control; Toxicity; hemolysis test; Biomaterial; Material testing; Biomedical engineering; guidance
• ISSN: 1552-4973; 1552-4981
• DOI: 10.1002/jbm.b.33169

This study deals with the development and performance evaluation of a positive reference material for hemolysis testing, which is used for evaluating the biological safety of medical devices. Genapol X-080, a nonionic detergent, was selected as a candidate hemolytic substance in a survey of 23 chemical compounds; it showed significant hemolytic activity against rabbit defibrinated blood at concentrations more than 20 µg/mL. A polyvinyl chloride (PVC) sheet spiked with 0.6% (w/w) of the compound exhibited weak hemolytic activity in direct contact and/or extract-based assays after 4 h incubation at 37°C. A PVC sheet containing 5.8% (w/w) Genapol X-080 induced complete hemolysis in both assays. The amount of Genapol X-080 eluted from each PVC sheet during hemolysis testing using the direct contact method increased time-dependently and reached 25.6 (former sheet) or 1154 (later sheet) µg/mL after 4 h incubation, which was similar to or much higher than the critical micelle concentration, respectively. Similar elution behavior was observed using the extract-based method, and the Genapol X-080 content in test solutions prepared by autoclave extraction of both sheets was 22.5 and 358 µg/mL, respectively, indicating a clear relationship between the degree of hemolytic activity and the eluted amount of Genapol X-080. Thus, a PVC sheet spiked with a compound exhibiting different hemolytic activity depending on its concentration may be useful as a positive reference material to validate the hemolysis tests.