p-vinylphenol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

It was concluded that PHS has mutagenic potential in L5178Y TK+/-− -clone 3.7.2C cells under the conditions of this study.

No in-vivo test is proposed since the monomer is not supplied into the EU only the polymer

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 January to 20 February 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian cell gene mutation tests using the thymidine kinase gene

Species / strain / cell type:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Metabolic activation system:

Short-term treatment in the absence of metabolic activation (−S9 mix) Short-term treatment in the presence of metabolic activation (+S9 mix): 24-hour continuous treatment (−S9 mix): 1.32, 1.98, 2.96, 4.44, 6.67, 10.0 and 15.0 μg/mL

Test concentrations with justification for top dose:

In accordance with the specification of “Toxicity Study Guidelines”, the highest dose level was set at 1200 μg/mL (equivalent to 10 mM), and this was diluted using a common ratio of 2 to obtain a total of 9 concentrations (600, 300, 150, 75.0, 37.5, 18.8, 9.38 and 4.69 μg/mL).

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

cyclophosphamide

methylmethanesulfonate

Key result

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

PHS has mutagenic potential in L5178Y TK+/-− -clone 3.7.2C cells under the conditions of this study.

Executive summary:

A dose range finding test and a gene mutation test were conducted for PHS using the mutations in the thymidine kinase gene loci in the cultured mouse lymphocytes (L5178Y TK+/− -clone 3.7.2C) cells with the short-term treatment and 24-hour continuous treatment.

Observation and measurement were performed after the treatment with the test article or the control articles for 3 hours for the short-term treatment and for 24 hours for the continuous treatment. Dimethyl sulfoxide (DMSO), the vehicle, was used as the negative control article and methyl methanesulfonate (MMS) and cyclophosphamide (CP) as the positive control articles for the treatment without and with metabolic activation, respectively.

Since cytotoxicity was occurred in the dose range finding test, the concentrations for the gene mutation test was selected to cover the cytotoxicity range from that producing cytotoxicity and including concentrations at which there is moderate and little or no cytotoxicity.

– Short-term treatment in the absence of metabolic activation (−S9 mix):

5.27, 7.90, 11.9, 17.8, 26.7, 40.0 and 60.0 μg/mL (common ratio: 1.5)

– Short-term treatment in the presence of metabolic activation (+S9 mix):

8.78, 13.2, 19.8, 29.6, 44.4, 66.7 and 100 μg/mL (common ratio: 1.5)

– 24-hour continuous treatment (−S9 mix):

1.32, 1.98, 2.96, 4.44, 6.67, 10.0 and 15.0 μg/mL (common ratio: 1.5)

In the judgment of the results, the indices calculated by addition of the Global Evaluation Factor (GEF: 126 × 10−6) to the total mutant frequency (T-MF) of the negative control group were used for evaluation according to recommendation of "OECD Guidelines for Testing of Chemicals 490".

In the gene mutation test, precipitation was not observed in any test article treatment group. Color of the culture medium did not change for any test article treatment group.

In the short-term treatment without metabolic activation, the T-MFs were 65.95, 132.75, 206.79, 314.11, 401.39, 800.00 and 871.09 × 10−6 at 5.27, 7.90, 11.9, 17.8, 26.7, 40.0 and 60.0 μg/mL, respectively, and the T-MFs at the dose level of 11.9 to 60.0 μg/mL were increased more than the indices calculated by addition of the GEF to the T-MF of the concurrent negative control group (73.63 × 10−6). However, the T-MFs at the dose levels of 40.0 and 60.0 μg/mL were excluded from the statistical analysis and the evaluation of the results, since the RTG was not higher than 10%. This treatment with the test article showed a significant dose-dependent

T-G327

9

T-MF increase (p < 0.05). According to the judging criteria, the result of the short-term treatment without metabolic activation was evaluated as positive.

In the short-term treatment with metabolic activation, the T-MFs were 88.95, 152.61, 205.27, 236.46, 302.97, 309.96, 413.79 × 10−6 at the dose levels of 8.78, 13.2, 19.8, 29.6, 44.4, 66.7 and 100 μg/mL, respectively, and the T-MFs at the dose levels of 19.8 to 100 μg/mL were increased more than the indices calculated by addition of the GEF to the T-MF of the concurrent negative control group (65.20 × 10−6). However, the T-MFs at the dose levels of 66.7 and 100 μg/mL were excluded from the statistical analysis and the evaluation of the results, since the RTG was not higher than 10%. This treatment with the test article also showed a significant dose-dependent T-MF increase (p < 0.05). According to the judging criteria, the result of the short-term treatment without metabolic activation was evaluated as positive.

In the continuous treatment, the T-MFs were 93.00, 62.97, 108.92, 98.93, 155.00, 246.84 and 288.02 × 10−6 at the dose levels of 1.32, 1.98, 2.96, 4.44, 6.67, 10.0 and 15.0 μg/mL, respectively, and the T-MFs at the dose levels of 10.0 and 15.0 μg/mL were increased more than the indices calculated by addition of the GEF to the T-MF of the concurrent negative control group (109.29 × 10−6). This treatment with the test article also caused a significant dose-dependent T-MF increase (p < 0.05). According to the judging criteria, the result of the continuous treatment without metabolic activation was evaluated as positive.

For the negative control group, the T-MF was within 50×10-6 to 170×10-6, the CE was within 0.65 to 1.20, and the TSG was between 8 to 32-fold in the short-term treatment and 32 to 180-fold in the continuous treatment. For the positive control group, the RTG was more than 10% and the S-MF increased 150×10-6 above the concurrent negative control group. For all the treatment groups, the number of evaluable dose levels was 4 or more. Therefore, it was judged that the study was conducted appropriately.

Based on the above results, it was concluded that PHS has mutagenic potential in L5178Y TK+/-− -clone 3.7.2C cells under the conditions of this study.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 to 11 September 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Maruzen Petrochemical Co. Ltd.; Bx 7706RXD
- Expiration date of the lot/batch: no data
- Purity test date: no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: The test item gradually polymerises at room temperature. Stable at storage conditions
- Stability under test conditions: no data
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

1250, 625, 313, 156, 78.1 and 39.1 µg/plate. The top dose was chosen because during dose selection testing bacterial growth inhibition was observed at 1250 µg/plate.

Vehicle / solvent:

- Vehicle used: DMSO (dehydrated)

- Justification for choice of vehicle: Because the test item gradually polymerises in water and acetone dehydrated DMSO was used.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

True negative controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide and 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable):

DURATION
- Preincubation period: no
- Exposure duration: 48h
- Expression time (cells in growth medium): 20min
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):

SPINDLE INHIBITOR (cytogenetic assays):

STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS: 2

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED:

NUMBER OF CELLS EVALUATED:

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells):

CRITERIA FOR MICRONUCLEUS IDENTIFICATION:

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:
- Any supplementary information relevant to cytotoxicity:

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable):

- OTHER:

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

It is concluded from this study that PHS had no ability to induce mutations under the test conditions.

Reference 3

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 October to 22 December 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Maruzen petrochemical Co. Ltd.; Lot no. 7706RXD
- Expiration date of the lot/batch: no data
- Purity test date: no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: stable
- Stability under test conditions: Readily polymerises
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

CELLS USED
- Source of cells: Commercial supplier
- Suitability of cells: recommended by test method
- Cell cycle length, doubling time or proliferation index: doubling time about 15 hours
- Sex, age and number of blood donors if applicable:
- Whether whole blood or separated lymphocytes were used if applicable:
- Number of passages if applicable:
- Methods for maintenance in cell culture if applicable:
- Modal number of chromosomes: 25 per cell
- Normal (negative control) cell cycle time:

MEDIA USED
- Type and identity of media including CO2 concentration if applicable:
- Properly maintained: [yes/no]
- Periodically checked for Mycoplasma contamination: [yes/no]
- Periodically checked for karyotype stability: [yes/no)
- Periodically 'cleansed' against high spontaneous background: [yes/no]

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

18.8, 37.5, 75.0, 150, 300, 600 and 1200 µg/L. 10 mmol/L i.e. 1200 µg/L was chosen as the maximum dose as this gave no cytotoxicity.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO;
- Justification for choice of solvent/vehicle: the substance polymerises at ambient.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium;
- Cell density at seeding (if applicable): 5xE03 cells/mL

DURATION
- Preincubation period: 3 days
- Exposure duration:
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):

SPINDLE INHIBITOR (cytogenetic assays):

STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS:

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED:

NUMBER OF CELLS EVALUATED:

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells):

CRITERIA FOR MICRONUCLEUS IDENTIFICATION:

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:
- Any supplementary information relevant to cytotoxicity:

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable):

- OTHER:

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not applicable

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Conclusions:

Under the conditions of this test it was concluded that PVP did not induce chromosomal aberrations.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)