Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 232-650-8 | CAS number: 9002-07-7
Trypsin was tested for genotoxicity potential using Ames and Chromosome Aberration test.
It was concluded that the results of the experiments give no indication of mutagenic activity of SP 387/TL1 in the presence or absence of metabolic activation, up to 5000 µg/mL under the conditions employed in this study.
It is concluded that SP 387/TL1 did not induce chromosome aberrations in cultured human peripheral blood lymphocytes when tested to 5000 µg/mL, an acceptable maximum concentration for chromosome aberration studies according to current regulatory guidelines, in both the absence and presence of a rat liver metabolic activation system (S-9).
SP 387/TL 1 (batch PPF26813) was examined for mutagenic activity in the bacterial reverse mutation assay using Salmonella typhimurium strain TA1535, TA100, TA1537, TA98 and Escherichia coli W P2uvr A. Crude enzyme preparations, like the present batch of SP387/TL 1, contain the free amino acid histidine, most often in an amount, which exceeds the critical concentration for incorporation in the direct standard assay. To overcome this problem all Salmonella typhimurium strains were exposed to SP387/TL 1 in liquid culture ("treat and plate assay"). Bacteria were exposed to 6 doses of the test substance in a phosphate buffered nutrient broth for 3 hours with 5 mg (dry matter) per ml as highest concentration. After incubation the test substance was removed by centrifugation prior to plating.
Usually the content of tryptophan in enzyme preparations is low and insignificant. Therefore the part of the study comprising Escherichia coli was conducted with the strain WP2uvrA using the direct plate incorporation assay. 6 doses of the test substance were applied with 5 mg (dry matter) per plate as the highest dose level followed by successive bi-sections between doses. The study was conducted with and without the metabolic activation system S9 - a liver preparation from male rats, pre-treated with Aroclor 1254, and the co-factors required for mixed function oxidase activity (S9 mix). Two identical and independent experiments were conducted.
A test substance was considered as positive when it had induced at least a doubling in the mean number of revertants per plate compared to the appropriate solvent control in one or more of the strains, in the presence or absence of S9, if this response was dose related and reproducible. If a dose-related numerical increased below a doubling but at least 50% higher than the solvent control was observed then the result was considered as equivocal and would need further clarification.
Cytotoxicity was observed at the highest concentration of 5000 μg substance/mL in experiment 1 in strain TA1537 in the presence of S9; and in experiment 2 in strains TA1537 and TA1535 in the presence of S9. No treatments of any of the Salmonella typhimurium and E. coli strains with SP 387/TL 1, batch PPF26813 either in the presence or absence of S-9 mix, resulted in any increases in revertant numbers that meets these criteria for a positive response.
It was concluded, that the results of the experiments, described in this report, gave no indication of mutagenic activity of SP 387/TL1, batch PPF26813 in the presence or absence of metabolic activation, when tested under the conditions employed in this study.
SP 387/TL1 was tested in an in vitro cytogenetics assay using duplicate human lymphocyte cultures prepared from the pooled blood of three female donors in two independent experiments. Treatments covering a broad range of concentrations, separated by narrow intervals, were performed both in the absence and presence of metabolic activation (S-9). The test article was formulated in sterile water for injection (purified water) and the highest concentration used, 5000 μg/mL, is an acceptable maximum concentration for in vitro chromosome aberration studies according to current regulatory guidelines. The chromosome aberration assay was used to evaluate the clastogenic potential of the test article.
In Experiment 1, treatment in the absence and presence of S-9 was for 3 hours followed by a 17 hour recovery period prior to harvest (3+17). The S-9 used was prepared from a rat liver post-mitochondrial fraction (S-9) from Aroclor 1254 induced animals. The test article concentrations for chromosome analysis were selected by evaluating the effect of SP 387/TL1 on mitotic index. Chromosome aberrations were analysed at three concentrations, 2813, 3750, 5000 μg/mL.
In Experiment 2, treatment in the absence of S-9 was continuous for 20 hours (20+0). Treatment in the presence of S-9 was for 3 hours only followed by a 17 hour recovery period prior to harvest (3+17). Chromosome aberrations were analysed at three concentrations 3200, 4000, 5000 μg/mL.
Appropriate negative (vehicle) control cultures were included in the test system in both experiments under each treatment condition. The proportion of cells with structural aberrations in these cultures fell within historical vehicle control ranges. 4-Nitroquinoline 1-oxide (NQO) and Cyclophosphamide (CPA) were employed as positive control chemicals in the absence and presence of rat liver S-9 respectively. Cells receiving these were sampled in each experiment, 20 hours after the start of treatment; both compounds induced statistically significant increases in the proportion of cells with structural aberrations. Treatment of cultures with SP 387/TL1 in the absence and the presence of metabolic activation (S-9) [both experiments] resulted in frequencies of cells with structural chromosome aberrations (excluding gaps), which were similar to those observed in concurrent vehicle control cultures for all concentrations analysed. The aberrant cell frequency of all SP 387/TL1 treated cultures fell within (or very close to) current historical vehicle control (normal) ranges. No increases in the frequency of cells with numerical aberrations, which exceeded the historical negative control range were observed in cultures treated with SP 387/TL1 in the absence and presence of S-9 (both experiments).
It was concluded that SP 387/TL1 did not induce chromosome aberrations in cultured human peripheral blood lymphocytes when tested to 5000 mg/mL, an acceptable maximum concentration for chromosome aberration studies according to current regulatory guidelines, in both the absence and presence of a rat liver metabolic activation system (S-9).
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
Welcome to the ECHA website. This site is not fully supported in Internet Explorer 7 (and earlier versions). Please upgrade your Internet Explorer to a newer version.
Close Do not show this message again