Development and performance evaluation of a positive reference material for hemolysis testing.
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.
Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan.
Yuji
Y
Haishima
Chie
C
Hasegawa
Yusuke
Y
Nomura
Tsuyoshi
T
Kawakami
Toshiyasu
T
Yuba
Tomoko
T
Shindo
Keisuke
K
Sakaguchi
Takahiro
T
Tanigawa
Kaori
K
Inukai
Mika
M
Takenouchi
Kazuo
K
Isama
Atsuko
A
Matsuoka
Shingo
S
Niimi
D000818:Animals; D006403:Hematologic Tests / Q000379:methods* / Q000592:standards*; D006461:Hemolysis; D008823:Micelles; D011092:Polyethylene Glycols / Q000737:chemistry*; D011143:Polyvinyl Chloride; D011817:Rabbits; D012015:Reference Standards
D008823:Micelles; D011092:Polyethylene Glycols; D011143:Polyvinyl Chloride; C116996:genapol X 080
biological safety evaluation; guidance; hemolysis test; positive control