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EC number: 232-577-1 | CAS number: 9001-05-2
The clastogenic and aneugenic activity of catalase was investigated in cultured human peripheral blood lymphocytes by effects on the frequency of micronuclei. Division of the lymphocytes was stimulated by adding phytohaemagglutinin to the cultures. Sets of duplicate cultures were exposed to the test substance for 3 hours in the absence and presence of metabolic activation (S-9 mix) and harvested 24 hours after the beginning of treatment (3+21 hour treatment). Additionally, a continuous 24-hour treatment in absence of S-9 mix was included with harvesting 48 hours after the beginning of treatment (24+24 hour treatment). The cultures were treated with cytochalasin-B after removal of the test substance to block cytokinesis. Three concentrations, covering an appropriate range of cytotoxicity, were selected for scoring of micronuclei. A minimum of 1000 cells per concentration were scored.
The proportion of binucleate cells with micronuclei in all cultures of the vehicle controls (purified water) was within the limits of the historical ranges. The positive controls induced statistically significant increases in the proportion of cells with micronuclei, demonstrating the sensitivity of the test procedure and the metabolic activity of the S-9 mix employed.
Treatment of cells with Catalase, PPX 28086 in the absence and presence of S-9 resulted in frequencies of MNBN cells which were generally similar to, and not significantly (p ≤ 0.05) higher than those observed in concurrent vehicle controls for the majority of concentrations analysed (all treatments). Exceptions (3000 μg/mL, 3+21 hour +S-9 treatment) and (500.0 μg/mL, 24+24 hour -S-9 treatment) where small but statistically significant increases were noted, were not considered of biological importance.
It is concluded that Catalase, PPX 28086 did not induce micronuclei in cultured human peripheral blood lymphocytes following treatment in the absence and presence of an aroclor induced rat liver metabolic activation system (S-9). Maximum concentrations analysed were either up to the recommended regulatory maximum of 5000 μg/mL (3+21 hour +/-S-9 treatments), or to 3000 μg/mL (24+24 hour –S-9 treatment), limited by toxicity, in accordance with current regulatory guidelines for the in vitro micronucleus assay.
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