Formaldehyde, polymer with benzenamine, hydrogenated

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test substance did not exhibit clastogenic potential and it showed no evidence of mutagenic activity when tested in three in vitro mutagenicity tests according to OECD guidelines 471, 473 and 476.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (GLP)

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

The indicator cell used for this study was the L5178Y mouse lymphoma cell line that is heterozygous at the TK locus (+/-). The particular clone (3.7.2C) used in this assay is isolated by ATCC (American Type Culture Collection), 0801 University Blvd., Manassas, VA 20110-2209, USA.
Stock cultures were obtained from ATCC, and master stocks were maintained in liquid nitrogen. Laboratory cultures were periodically checked for karyotype stability and the absence of mycoplasma contamination by culturing methods. To reduce the background mutant frequency (spontaneous frequency) of TK-/- mutants to a level as low as possible, cell cultures were exposed to conditions that select against the TK-/- phenotype (exposure to aminopterin or methotrexate). Cell cultures were maintained in cleansing medium for one day, placed in recovery medium for one day and then returned to normal growth medium for three to eight days before use.

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

In the first experiment without metabolic activation and both experiments with metabolic activation (3-hour exposure):
15.63, 31.25, 62.5, 125 and 250 µg/mL medium
In the second experiment without metabolic activation (24-hour exposure):
7.81, 15.63, 31.25, 62.5 and 125 µg/mL medium.

Vehicle / solvent:

The test item was completely dissolved in dimethylsulfoxide (DMSO).
As recommended by the guidelines at least 10 exp5 cells were suspended in treatment medium and diluted to 5 x 10 exp5 cells/mL.
Fresh preparations of the test and reference item solutions were prepared on each day of biological testing.

Untreated negative controls:

no

Remarks:

The vehicle dimethylsulfoxide (DMSO) served as the negative control.

Negative solvent / vehicle controls:

yes

Remarks:

The vehicle dimethylsulfoxide (DMSO) served as the negative control.

True negative controls:

no

Positive controls:

yes

Positive control substance:

3-methylcholanthrene

methylmethanesulfonate

Remarks:

The vehicle dimethylsulfoxide (DMSO) served as the negative control.

Key result

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Conclusions:

Under the present test conditions, Copolymer of aniline and formaldehyde, hydrogenated, tested up to cytotoxic concentrations, in the absence and presence of metabolic activation in two independent experiments was negative with respect to the mutant frequency in the L5178Y TK +/- mammalian cell mutagenicity test. Under these conditions the positive controls exerted potent mutagenic effects and demonstrated the sensitivity of the test system and conditions.
In addition, no change was noted in the ratio of small to large mutant colonies. Therefore, Copolymer of aniline and formaldehyde, hydrogenated also did not exhibit clastogenic potential at the concentration-range investigated.
According to the evaluation criteria for this assay, these findings indicate that Copolymer of aniline and formaldehyde, hydrogenated, tested up to cytotoxic concentrations in the absence and presence of metabolic activation, did neither induce mutations nor have any chromosomal aberration potential.

Executive summary:

Copolymer of aniline and formaldehyde, hydrogenatedwas assayed in a gene mutation assay in cultured mammalian cells (L5178Y TK +/-) both in the presence and absence of metabolic activation by a liver post-mitochondrial fraction (S9 mix) from Aroclor 1254-induced rats. The test was carried out employing two exposure times without S9 mix: 3 and 24 hours, and one exposure time with S9 mix: 3 hours; the experiment with S9 mix was carried out in two independent assays.

Copolymer of aniline and formaldehyde, hydrogenated was completely dissolved indimethylsulfoxide (DMSO).The vehicle DMSO served as the negative control.

In the preliminary experiment without and with metabolic activation (24-hour or 3-h exposure)cytotoxicity (decreased survival) was noted starting at a concentrations of 100 or 250 µg/mL, respectively.

Hence, in the main studythe concentration-range of 15.63 to 250 µg/mL was usedin theexperiments without and with metabolic activation with a 3-hour exposure time and a concentration-range of 7.81 to 125 µg/mL was used in the second experiment without metabolic activation (24-hour exposure).

Methylmethanesulfonate (at 10 or 15 µL/mL) was employed as a positive control in the absence of exogenous metabolic activation and 3 -Methylcholanthrene (at 2.5 or 4.0 µg/mL) in the presence of exogenous metabolic activation.

In the main study,cytotoxicity(decreased survival) was noted in the presence and absence of metabolic activation (3-hour exposure) at the top concentration of 250 µg/mL and in the second experiment without metabolic activation (24-hour exposure) at the top concentration of 125 µg/mL.

The values of mutation frequencies of the negative controls ranged from 57.54 to 74.74 per 10⁶ clonable cells in the experiments without metabolic activation and from 55.50 to 69.29 per 10⁶ clonable cells in the experiments with metabolic activationand, hence, were well within the historical data-range.

The mutation frequencies of the cultures treated with Copolymer of aniline and formaldehyde, hydrogenated ranged from 67.37to 127.28 per 10⁶ clonable cells (3 hours exposure) and from 49.44 to 60.19 per 10⁶clonable cells (24 hours exposure) in the experiments without metabolic activation and from 63.68 to 95.20 per 10⁶clonable cells (3 hours exposure, first assay) and from 66.66 to 96.51 per 10⁶ clonable cells (3 hours exposure, second assay) in the experiments with metabolic activation. These results were within the range of the negative control values and the normal range of 50 to 170 mutants per 10⁶ viable cells and, hence, no mutagenicity was observed according to the criteria for assay evaluation.

In addition, no change was observed in the ratio of small to large mutant colonies, ranging from 0.43 to 1.50 for Copolymer of aniline and formaldehyde, hydrogenated treated cells and from 0.67 to 0.95 for the negative controls.

The plating efficiency (PE step 1 and step 2) of the negative control was ≥ 50%, the mean cloning efficiencies (CE) within the range of 65% to 120% two days after treatment, and the mean suspension growth (SG) within the range of 8 to 32 following 3 -hour treatments and between 32 and 180 following 24-hour treatments.

The positive controls Methylmethanesulfonate (MMS) and 3-Methylcholanthrene (3-MC) caused pronounced increases in the mutation frequency ranging from 1157.15 to 2592.61 per 10⁶ clonable cells in the case of MMS and ranging from 917.29 to 1434.11 per 10⁶ clonable cells in the case of 3 -MC.

In addition, the colony size ratio was moderately shifted towards an increase in small colonies, ranging from 1.65 to 2.29 in the case of MMS.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (GLP)

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

other: Chinese hamster lung (CHL) cells, strain JCRB0030

Details on mammalian cell type (if applicable):

JCRB0030
Chinese hamster lung (CHL) cells, strain JCRB0030, were obtained from SafePharm on 29 May 1995 at a passage number of 5 and stored in polypropylene ampoules at -196°C in 90% foetal calf serum and 10% dimethylsulphoxide. When the cells were first grown at Huntingdon Research Centre from those stored at -196 °C, they were given a passage number of 1. The passage numbers quoted in this report refer to those used once the cells were grown at HRC. The cells were routinely grown and subcultured in Minimal Essential Medium (MEM, Gibco) supplemented with 10% foetal calf serum (FCS, Imperial Laboratories) at 37°C in a humid atmosphere containing 5% carbon dioxide in 175 cm³ plastic tissue culture flasks (Falcon). The doubling time of this strain of CHL cells is approximately 12 hours and they have a modal chromosome number 25.

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S-9

Test concentrations with justification for top dose:

Initially cells were establisecl for set 4 ancl dosed as explained below to give final concentrations of
MPCA as follows:
39.1, 78.1, 156.3, 312.5, 625, 1250, 2500 and 5000 µg/ml

Vehicle / solvent:

DMSO
Prior to commencing testing, the solubility of the test substance in dimethyl sulphoxide was assessed.
MPCA was found to be soluble in DMSO at 500 mglml. This gave a tinal concentration of
5000 pglml when dosed at 1% vlv into tissue culture medium. 5000 pglml is the highest
concentration used in the test system of this nature as artefactual increases in chromosomal aberrations
may be caused by solutions of high ionic strength and osmolality (Galloway et al.). At 5000 µg/ml
a precipitate was formed in the culture medium. This was used as the highest dose level for
subsequent testing. in accortlance with Japanese guiclelines.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

other: Carbendazim

Details on test system and experimental conditions:

A 50 ml culture of CHL cells was harvested as follows: the supernatant medium was removed and
20 ml of 0.1 % trypsin was then added for 45 seconds. The trypsin solution was removed and the
flask incubated at 37°C for 10 minutes. The cells were then resuspended in 20 ml MEM + 10%
FCS and diluted to give 8 x 10 4 cells/ml for cultures undergoing a 24 hour incubation after addition
of test substance and 2.5 x l0 4 cells/ml for cultures to be incubated for 48 hours after addition of
test substance. Aliquots (5 ml) of cells were added to Nunc 25 cm' tissue culture flasks and the
cultures incubated at 37 °C in rt humid atmosphere containing 5% carbon dioxide.
After approximately 24 hours the cells were treated with the test substance. Four sets of treatments
were used which are summarised as follows:
Set 1: 6 hours treatment, +S-9, harvest at 24 hours.
Set 2: 6 hours treatment, -S-9, harvest at 24 hours.
Set 3: 24 hours treatment, -S-9, harvest at 24 hours.
Set 4: 48 hours treatment, -S-9, harvest at 48 hours.
The highest concentration of test substance had been prepared previously by dissolving in DMSO.
This was diluted so that when dosed at 10 µl/ml the final concentrations would be as follows
Set 1 : 2.5, 5, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 and 4246 µg/ml
Set 2: 0.625, 1.25, 2.5, 5, 10, 20, 40, 80, 160, 320. 640, 1280 µg/ml
Set 3: 0.625, 1.25, 2.5, 5, 10, 20, 40, 80, 160, 320 µg/ml
Set 4: 0.157, 0.313, 0.625, 1.25, 2.5, 5, 10, 20, 40, 80 µg/ml
To the tirst set of cultures 1.25 ml of S-9 mix was added followed by 62.5 µl of the various dilutions
of MPCA. Two cultures were treated with each dose level. Four cultures were treated with 62.5 µl
of the solvent control, DMSO, and two cultures with 62.5 µl of the clastogenic positive control
compound, cyclophosphamide, at final concentrations of 10 and 15 µg/ml.
To the second, third and fourth sets of cultures, without S-9 mix, 50 µl aliquots of the dilutions of
MPCA were added so that two cultures were treated with each dose level. 50 µl of the solvent
control was added to four cultures in each set and 50 µl aliquots of the following positive control
compounds were added to two cultures in each set. The clastogenic positive control, mitomycin C,
at a final concentration of 0.2 and 0.4 µg/ml was added to cultures in set 2, and 0.1 and 0.2 µg/ml
to cultures in sets 3 and 4. Carbendazim, 50% w/w, the positive control for polyploidy, was added
to cultures in set 2 at final concentrations of 12 and 24 µg/ml, to set 3 at final concentrations of 6 and
12 µg/ml and to set 4 at final concentrations of 3 and 6 µg/ml.
The first three sets of cultures were then incubated at 37°C for 24 hours and the fourth set of cultures
for 48 hours.
Six hours after addition of the test compound to the first and second sets of cultures, the medium
containing the test substance was removed and replaced with fresh MEM + 10% FCS. The cultures
were returned to the incubator for a further 18 hours.

HARVESTING AND FIXATION
Two hours before the end of the 24 hour incubation period mitotic activity was arrested in the first
three sets of cultures by the acldition of Colcemid (Sigma) to each culture at a final concentration
of 0.1 µg/ml. Colcemid (0.1 µg/ml) was added to the fourth set of cultures two hours before the
end of their 48 hour incubation period.
After the incubation period the medium was removed and retained. 4 ml of 0.1 % trypsin solution
was then added. After one minute this was removed and placed in a plastic conical centrifuge tube,
with the medium that had been removed. The flasks were then incubated for 5 minutes at 37°C after
which the cells were resuspentled in the combined medium and trypsin solution. The cell suspensions
were placed with the trypsin solution in the centrifuge tubes. These cell suspensions were then
centrifuged for 10 minutes at 200 g. The supernatant was discarded and the cells resuspended in 2.5
ml 0.07M KCI. After a 10 minute incubation at room temperature the cell suspensions were
centrifuged for 5 minutes at 110 g. The supernatant was discarded and 4 ml of freshly prepared
fixative (3 parts methanol : 1 part glacial acetic acid v/v) added. The pellets were allowed to fix for
at least 2 hours.
SLIDE PREPARATION
The pellets were resuspendetl by repeated aspiration through a 20 gauge needle, centrifuged at 200 g
for 10 minutes, the supernatant discarded, and the cell pellet resuspended in a small volume of fresh
fixative. Two or three drops of this cell suspension were dropped onto cold, pre-cleaned microscope
slides which were then allowed to air-dry. The slides were then placed in buffered distilled water
(pH 6.8) prior to staining in Giemsa (dilution factor : 1 part Giemsa to 9 parts buffered distilled
water). After rinsing in hufferecl clistilled wates the slides were left to air-dry and then mounted in
DPX.
MlCROSCOPIC EXAMINATION
The prepared slides were examined at a magnification of x160. The proportion of mitotic cells per
1000 cells in each culture was recorded except for positive control treated cultures. From these results at least three dose levels for subsecluent metaphase analysis were selected for each treatment
regime. The highest dose level was that causing a depression in mitotic index of approximately 50%
when compared with the solvent control level. Intermecliate and low concentrations usually
approximated to 50% and 25% of the highest close level.
The concentration of positive control compounds selected for analysis was the lowest dosed unless
a preliminary scan of metaphase figures inclicatecl an insufticient level of aberrant cells.
Prior to metaphase analysis all slides were coded. Metaphase figures were identified, using a
magnification of x160, and examined at a magnification of x1000 using an oil immersion objective.
100 metaphase figures were examined, where possible, from each culture. Only cells with 23 - 27
chromosomes were analysed for structural aberrations. The vernier readings of all aberrant metaphase
figures were recorded. Additionally, the proportion of polyploid cells in 100 metaphase figures,
where possible, was recorded.
The number of aberrant metaphase figures, or polyploid cells, in each treatment group was compared
with the solvent control value using Fisher's test (Fisher 1973).

Species / strain:

mammalian cell line, other: Chinese hamster lung

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: strain/cell type: Chinese hamster lung cells

Remarks:

Migrated from field 'Test system'.

Conclusions:

Formaldehyde, polymer with benzenamine, hydrogenated has shown no evidence of clastogenic activity in this in vitro cytogenetic test system.
However, polyploidy inducing activity was seen in the absence of S-9 mix, with some evidence of
polyploidy-inducing activity in its presence.

Executive summary:

With S-9 mix, 6 hour treatment, harvest at 24 hours At 40 µg/ml there was a reduction in the mitotic index to 53% of the solvent control value. The higher concentration was too toxic for analysis. The dose levels chosen for analysis were 10, 20 and 40 µg/ml. The concentration of cyclophosphamide selected for analysis was 15 µg/ml. Set 2: Without S-9 mix, 6 hour treatment, harvest at 24 hours At 20 µg/rnl there was a reduction in the mitotic index to 38% of the solvent control value. This dose level together with 5 and 10 µg/ml were chosen for analysis. The concentrations of mitomycin C and carbendazim selected for analysis were 0.4 and 24 µg/ml, respectively. Set 3: Without S-9 mix, 24 hour treatment and harvest at 5 µg/ml the mitotic index was reduced to 50% of the solvent control value. The higher dose level of 10 µg/ml was too toxic for analysis. 1.25, 2.5 and 5 µg/ml were chosen for analysis. The concentrations of mitomycin C and carbendazim selected for analysis were 0.2 and 6 µg/ml respectively. Set 4: Without S-9 mix, 48 hour treatment aod harvest The mitotic index was reduced to 31 % of the solvent control value at 5 µg/ml. This dose level together with 1.25 and 2.5 µg/ml were chosen for analysis. The concentrations of mitomycin C and carbendazin chosen for analysis were 0.1 and 3 µg/ml, respectively. No statistically significant increases in the proportion of aberrant cells were seen in any treatment set in the absence or the presence of S-9 mix. There were no statistically signiticant increases in the proportion of polyploid cells seen in sets 3 and 4. However, statistically signiticant increases were seen at the highest dose level in sets 1 and 2. The increase in set one. to 5.5%, was within the historical control range of 0.25-6.75%. The increase in set 2, to 14% at the top close level of 20 µg/ml, falls well outside the historical control value of 7% and therefore is considered to be treatment-related. All positive control compounds caused large, statistically signiticant increases in either the proportion of aberrant cells or the proportion of polyploid cells. This demonstrates the efficacy of the S-9 mix and the sensitivity of the test system.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (GLP)

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

other: histidine dependent auxotrophic mutants of Salmonella qphimurium (strains TA 1535, TA 1537, TA 98 and TA 100) and a tryptophan dependent mutant of Escherichia coli (WP2 uvrA)

Metabolic activation:

with and without

Metabolic activation system:

S-9

Test concentrations with justification for top dose:

In the preliminary toxicity test with dose levels of up to 5000 µg/plate some toxicity was observed
at the top dose level. A top dose level of 5000 µg/plate was chosen for the subsequent mutation
study. Other dose levels used in the mutation assays were: 2500, 1250, 625, 3 12.5, 156.25 and
78.13 µg/plate.

Vehicle / solvent:

Prior to commencing testing the solubility of the test substance was assessed at 50 mglml in water
and dimethyl sulphoxide. At this concentration the test substance was found to be immiscible with
water but fully miscible with dimethyl sulphoxide. Therefore dimethyl sulphoxide was chosen as the
solvent for this study.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

other: 2-Aminoanthracene

Details on test system and experimental conditions:

The test substance was added to cultures of the five tester strains at seven concentrations separated
by 2-fold dilutions. The highest concentration of MPCA used was 5000 pglplate. The negative
control was the chosen solvent, dimethyl sulphoxide. The positive control compounds were also
included.
An aliquot of 0.1 ml of a 10 hour bacterial culture and 0.5 ml S-9 mix or 0.5 ml 0.1 M phosphate
buffer (pH 7.4) were placed in glass bottles. An aliquot of 0.1 ml of the test solution was added,
followed immediately by 2 ml of histidineltryptophan deficient agar. The mixture was thoroughly
shaken and overlaid onto previously prepared petri dishes containing 25 ml minimal agar. Three petri
dishes were used for each dose level. A set of plates were also prepared containing only bacterial
culture and S-9 mix or phosphate buffer (0 pglplate). Plates were also prepared without the addition
of bacteria in order to assess the sterility of the test substance, S-9 mix and phosphate buffer. All
plates were incubated at 37°C for 3 days. After this period revertant colonies were counted using
a Seescan Automatic Colony Counter.
At a later date the main test was repeated using the procedures described above with the same
concentrations of test substance.

Evaluation criteria:

The mean number of revertant colonies for all treatment groups is compared with those obtained for
solvent control groups. The mutagenic activity of a test substance is assessed by applying the
following criteria:
(a) If treatment with a test substance produces an increase in revertant colony numbers of at least
twice the concurrent solvent controls, with some evidence of a positive dose-relationship, in
two separate experiments, with any bacterial strain either in the presence or absence of S-9
mix, it is considered to show evidence of mutagenic activity in this test system. No statistical
analysis is performed.
(b) If treatment with a test substance does not produce reproducible increases of at least 1.5 times
the concurrent solvent controls, at any dose level with any bacterial strain, it is considered to
show no evidence of mutagenic activity in this test system. No statistical analysis is performed.
(c) If the results obtained fail to satisfy the criteria for a clear "positive" or "negative" response
given in paragraphs (a) and (b), the following approach is taken in order to resolve the issue
of the substance's mutagenic activity in this test system.
(i) Repeat tests may be performed using modifications of the experimental method. These
modifications include (but are not restricted to), the use of a narrower dose range than
that already tested; the use of different levels of liver homogenate S-9 fraction in the S-9
mix. Should an increase in revertant colony numbers be observed which satisfies
paragraph (a) the substance is considered to show evidence of mutagenic activity in this
test system. No statistical analysis is performed.
(ii) If no clear "positive" response can be obtained the test data may be subjected to analysis
to determine the statistical significance of any observed increases in revertant colony
numbers. The statistical procedures used will be those described by Mahon et al.
(1989).

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Conclusions:

Test substance, when tested in dimethyl sulphoxide, up to cytotoxic concentrations in the presence or absence of metabolic activation showed no evidence of mutagenic activity in this bacterial reverse mutatin assay.

Executive summary:

The test substance was toxic at 5000 µg/plate towards all the tester strains in the absence of S-9 mix and towards TA 98 and TA 100 in the presence of S-9 mix. Therefore 5000 µg/plate was chosen as the top dose level in the first mutation test but a total of seven dose levels were used to ensure that sufficient non toxic concentrations were assessed. Toxicity was observed at both 5000 and 2500 µg/plate but as five nontoxic dose levels were obtained it was decided that the same concentrations should be used in the second mutation test. No substantial increases in revertant colony numbers of any of the tester strains were observed following treatment with the test substance at any dose level, in the presence or absence of S-9 mix, in either mutation test. The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Additional information from genetic toxicity in vitro:

Formaldehyde, polymer with benzenamine, hydrogenated (MPCA) when tested in dimethyl sulphoxide, up to cytotoxic concentrations in the presence or absence of metabolic activation showed no evidence of mutagenic activity in this bacterial reverse mutation assay according to OECD guideline 471/2.

MPCA tested up to cytotoxic concentrations, in the absence and presence of metabolic activation in two independent experiments was negative with respect to the mutant frequency in the L5178Y TK +/- mammalian cell mutagenicity test according to OECD guideline 476. In addition, no change was noted in the ratio of small to large mutant colonies. Therefore, MPCA also did not exhibit clastogenic potential in this test.

 

MPCA tested up to cytotoxic concentrations, in the absence and presence of metabolic activation showed no evidence of clastogenic activity in the in vitro cytogenetic test system according to OECD 473 Guideline.  However, polyploidy inducing activity was seen in the absence of S-9 mix, with some evidence of polyploidy-inducing activity in its presence.

Justification for selection of genetic toxicity endpoint
Most recent study report (2013) supported by two other studies from 1995. All three reports are done according to EC Directive 92/69/EEC and Regulation EC/440/2008 guideline methods under GLP conditions.

Justification for classification or non-classification

Based on the results of three in vitro mutagenicity studies and according to the criteria of CLP Regulation 1272/2008 the test substance must not be classified.