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EC number: 948-912-6
CAS number: -
Gene mutation in bacteria: non mutagenic
Gene mutation in mammalian cells: non
According to Annex VII of the REACH
Regulation (EC 1907/2006) and ECHA Guidance Chapter R.7a: Endpoint
specific guidance, Version 5.0 – December 2016, a preliminary assessment
of mutagenicity should normally include data from a gene mutation test
in bacteria unless existing data for analogous substances indicates that
this would be inappropriate. When the result of the bacterial test is
positive, it is important to consider the possibility of the substance
being genotoxic in mammalian cells, whereas a negative response would
generally not require any further testing. However, if results from
further studies of genotoxicity are available, it is recommended to take
them into account in the assessement.
No data on target substance was
available, thus a read across approach was followed using available data
on Similar Substance 01 and Similar Substance 02. Details on the read
across are available in section 13 .
In particular, the assessment was
based on available data on mutagenic potential of Similar Substance 01
and 02 in Ames assay (OECD 471) as well as on mutagenic potential of
Similar Substance 01 in mammalian cells (OECD 476).
Both Ames assays on Similar
Substance 01 included 4 Salmonella typhimurium strains instead of the 5
recommended by the current version of OECD guideline 471. However,
significance and reliability of the result was confirmed by an available
study on Similar Substance 02, which included the 5 required strains.
In an Ames assay from 1995 on
Similar Substance 01, a sample with high purity (90 % dye, described as
"purified") was tested at concentration up to 5000 µg/plate using water
as vehicle. The study included a plate
incorporation test (experiment I) and a pre-incubation test (experiment
II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98,
and TA 100.
Slight toxic effects occurred in strains
TA 1535 and TA 1537 without S9 mix.
Slight increases in revertant colony
numbers were observed in strain TA 1537 at 5000.0 µg/plate without S9
mix and at 2500.0 µg/plate with S9 mix as well as in strain TA 98 at
5000.0 µg/plate with S9 mix in experiment II. These effects were
considered as biologically irrelevant, due to the relatively low level
of of the corresponding solvent control (strain TA 1537) and could not
be reproduced in the independent experiment (strain TA 1537 and TA 98).
On these bases, test substance was
considered as non-mutagenic in this assay.
In an Ames assay from 1994 on
Similar Substance 01, a sample with 80 % purity, described as "raw", was
tested at concentration up to 5000 µg/plate using DMSO as vehicle. Two
independent experiments were run using the plate incorporation method.
In the original and confirmatory
experiment, strain TA 1537 without metabolic activation showed a slight
increase in the number of revertant colonies at the concentration of
5000 µg/plate. No effect was seen with TA 1537 with metabolic
activation. Moreover, no effect was seen in the other strains both with
and without metabolic activation.
As no mutagenic effect was seen in
the study from 1995, despite
- the content of test substance was
higher than in the test sample that used for the study from 1994,
- both plate incorporation and
preincubation method were used,
the slight mutagenic response in the
study from 1994 was reasonably attributed to the impurities content of
test sample. Therefore, such study was disregarded, while the study from
1995 was selected as key study.
In addition, an Ames assay was
conducted on Similar Substance 02 using 5 Salmonella typhimurium
- TA 1535, TA 100 and TA 102 to
detect base-pair substitutions,
- TA 98 and TA 1537 to detect
No cytotoxicity, evident as
reduction in the number of revertants, was reported. Moreover, no
mutagenic effects were noted, in both the original and the confirmatory
experiment; in particular, positive findings with TA 102 strain in the
confirmatory experiment were considered as incidental.
Based on these experimental
evidences, a mutagenic potential was reasonably excluded.
A gene mutation assay in mammalian
cells with Similar Substance 01 was run using a sample of 80 % purity,
i.e. same sample as that exhibiting a mutagenic effect in the Ames assay
from 1994. Two independent experiments were run and each concentration
was tested in duplicate. Test concentrations were selected in a
preliminary test, where cytotoxic effects were noted. In both
experiments, no relevant increase of mutant frequencies compared to
controls was noted. Accordingly, a mutagenic potential in this assay was
According to the CLP Regulation (EC
1272/2008), a mutation means a permanent change in the amount or
structure of the genetic material in a cell. The term ‘mutation’ applies
both to heritable genetic changes that may be manifested at the
phenotypic level and to the underlying DNA modifications when known.
The more general terms ‘genotoxic’
and ‘genotoxicity’ apply to agents or processes which alter the
structure, information content, or segregation of DNA, including those
which cause DNA damage by interfering with normal replication processes,
or which in a non- physiological manner (temporarily) alter its
For the purpose of classification
for germ cell mutagenicity, substances are allocated to one of two
Category 1: substances known to
induce heritable mutations or to be regarded as if they induce heritable
mutations in the germ cells of humans.
Category 1A: based on positive
evidence from human epidemiological studies.
Category 1B: based on:
- positive result(s) from in vivo
heritable germ cell mutagenicity tests in mammals; or
- positive result(s) from in vivo
somatic cell mutagenicity tests in mammals, in combination with some
evidence that the substance has potential to cause mutations to germ
- positive results from tests
showing mutagenic effects in the germ cells of humans, without
demonstration of transmission to progeny; for example, an increase in
the frequency of aneuploidy in sperm cells of exposed people.
Category 2: substances which cause
concern for humans owing to the possibility that they may induce
heritable mutations in the germ cells of humans, based on positive
evidence obtained from experiments in mammals and/or in some cases from
in vitro experiments, obtained from:
- somatic cell mutagenicity tests in
vivo, in mammals; or
- other in vivo somatic cell
genotoxicity tests which are supported by positive results from in vitro
Based on negative result in
available experimental studies, a mutagenic potential was excluded and
the substance was not classified under the CLP Regulation (EC 1272/2008).
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.
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