Triphenylphosphine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

A bacterial reverse mutation assay revealed no mutagenic potential of the test item in 4 strains of S. typhimurium (Zeiger et al., 1987).

A micronucleus test in Chinese Hamster Lung cells in vitro was negative (Ni et al., 1993).

The test item did not induce gene mutations at the HPRT locus in a mammalian cell in vitro test according to OECD TG 476 and GLP (BASF, 2014).

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-Dec-05 - 2014-May-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5300 - In vitro Mammalian Cell Gene Mutation Test

Qualifier:

according to guideline

Guideline:

EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wsserwirtschaft und Gewerbeaufsicht, Kaiser-Friedrich-Straße 7, D-55116 Mainz

Type of assay:

mammalian cell gene mutation assay

Specific details on test material used for the study:

- Purity: 99.71%

SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 130034P040.
- Expiration date of the lot/batch: 05 Jul 2015.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature.
- Stability under test conditions: The stability of the test substance at room temperature in the vehicle THF over a period of 4 hours was determined analytically. The test substance was stable for 2 hours, after 4 hours a recovery rate of about 88% was obtained. However, all test substance solutions were prepared immediately before administration.
- Solubility and stability of the test substance in the solvent/vehicle: Due to the insolubility of the test substance in several commonly used vehicles, tetrahydrofurane (THF) was selected as vehicle.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The substance was dissolved in tetrahydrofurane (THF).

FORM AS APPLIED IN THE TEST (if different from that of starting material): solution in in tetrahydrofurane (THF).

Target gene:

HPRT locus

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

- Type and identity of media:
Treatment medium (with S9 mix): Ham's F12 medium containing stable glutamine and hypoxanthine;
Treatment medium (without S9 mix): Ham's F12 medium containing stable glutamine and hypoxanthine supplemented with 10% (v/v) fetal calf serum (FCS)
Culture medium: Ham's F12 medium containing stable glutamine and hypoxanthine supplemented with 10% (v/v) fetal calf serum (FCS);
Pretreatment medium ("HAT" medium): Ham's F12 medium supplemented with hypoxanthine (13.6 x 10E-3 mg/mL), aminopterin (0.18 x 10E-3 mg/mL), thymidine (3.88 x 10E-3 mg/mL), 10% (v/v) fetal calf serum (FCS);
Selection medium ("TG" medium): Hypoxanthine-free Ham's F12 medium supplemented with 6-thioguanine (10 μg/mL), 1% (v/v) stable glutamine (200 mM), 10% (v/v) fetal calf serum (FCS).
All media were supplemented with 1% (v/v) penicillin/streptomycin (stock solution: 10000 IU / 10000 μg/mL) and 1% (v/v) amphotericine B (stock solution: 250 μg/mL)

- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes (Each batch used for mutagenicity testing was checked for mycoplasma contamination)
- Periodically "cleansed" against high spontaneous background: yes

Metabolic activation:

with and without

Metabolic activation system:

phenobarbital/β-naphthoflavone-induced rat liver S9 mix

Test concentrations with justification for top dose:

1st Experiment with S9 mix: 6.25; 12.50: 25.00; 50.00; 100.00; 200.00 µg/mL
1st Experiment without S9 mix: 2.5; 5.00; 10.00: 20.00; 40.00; 80.00 µg/mL
2nd Experiment with S9 mix: 3.13; 6.25; 12.50; 25.00; 50.00; 100.00 µg/mL
2nd Experiment without S9 mix: 0.78; 1.56; 3.13; 6.25; 12.50; 25.00; 50.00; 100.00 µg/mL
3rd Experiment without S9 mix: 0.63; 1.25; 2.50; 5.00; 10.00; 20.00; 40.00 µg/mL

Following the requirements of the current international guidelines and the ICPEMC Task Group (4) a test substance should be tested up to a maximum concentration of 5 mg/mL, 5 μL/mL or 10 mM, whichever is the lowest. In case of toxicity, the top dose should result in approximately 10 - 20% relative survival (relative cloning efficiency), but not less than 10%. For relatively insoluble test substances at least one concentration should be scored showing no precipitation in culture medium at the end of the exposure period.
In the pretest for toxicity based on the purity and the molecular weight of the test substance 2600 μg/mL (approx. 10 mM) test substance was used as top concentration both with and without S9 mix at 4-hour exposure time.
In the pretests the pH value was not relevantly influenced by the addition of the test substance preparation to the culture medium at the concentrations measured. In addition, precipitation of the test substance in the vehicle tetrahydroflurane was not
observed in the stock solution (Test group: 2600 μg/mL). Test substance precipitation (macroscopical detection) occurred by the end of treatment at concentrations of 325.0 μg/mL and above in the absence and presence of S9 mix.
After 4 hours treatment in the absence of S9 mix, cytotoxicity was observed as indicated by a reduced relative cloning efficiency of about or below 20% relative survival at 162.5 μg/mL and above. In addition, in the presence of S9 mix, a clearly reduced relative cloning efficiency was observed after treatment with 40.6 μg/mL and above.

Vehicle / solvent:

- Vehicle/solvent used: tetrahydrofurane (THF)
- Justification for choice of solvent/vehicle: Due to the insolubility of the test substance in several commonly used vehicles, tetrahydrofurane (THF) was selected as vehicle,which has been demonstrated to be suitable in the CHO/HPRT assay and for which historical contral data are available.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

ethylmethanesulphonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium.

DURATION
- Exposure duration: 4 h (with and without S9).
- Expression time (cells in growth medium): 7 - 9 days.
- Selection time (if incubation with a selection agent): 6 - 7 days.
- Fixation time (start of exposure up to fixation or harvest of cells): 15 days.

SELECTION AGENT (mutation assays): 6-thioguanidine .
STAIN (for cytogenetic assays): Giemsa.

NUMBER OF REPLICATIONS: Duplicate cultures were used for all experimental groups.

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency.

Evaluation criteria:

ACCEPTANCE CRITERIA:
The HPRT assay is considered valid if the following criteria are met:
- The absolute cloning efficiencies of the negative/vehicle controls should not be less than 50% (with and without S9 mix).
- The background mutant frequency in the negative/vehicle controls should fall within our historical negative control data range of 0.00 – 16.43 mutants per 10E+06 clonable cells.
- The positive controls both with and without S9 mix must induce distinctly increased mutant frequencies (historical positive control data).
- At least 4 dose levels ranging up to a toxic concentration or up to or beyond the limit of solubility under culture conditions should be tested. Freely soluble and apparently non-toxic substances are not tested at concentrations higher than 5 mg/mL or 10 mM.

ASSESSMENT CRITERIA:
A finding is assessed as positive if the following criteria are met:
- Increase of the corrected mutation frequencies (MF[corr.]) both above the concurrent negative control values and our historical negative control data range.
- Evidence of reproducibility of any increase in mutant frequencies.
- A statistically significant increase in mutant frequencies and the evidence of a dose-response relationship.
Isolated increases of mutant frequencies above our historical negative control range (i.e. 15 mutants per 10E+06 clonable cells) or isolated statistically significant increases without a dose-response relationship may indicate a biological effect but are not regarded as sufficient evidence of mutagenicity.
The test substance is considered non-mutagenic according to the following criteria:
- The corrected mutation frequency (MF[corr.]) in the dose groups is not statistically significant increased above the concurrent negative control and is within our historical negative control data range.

Statistics:

An appropriate statistical trend test was performed to assess a dose-related increase of mutant frequencies. The number of mutant colonies obtained for the test substance treated groups was compared with that of the respective negative control groups. A trend is judged as statistically significant whenever the p-value (probability value) is below 0.05 and the slope is greater than 0. However, both, biological and statistical significance will be considered together.

Species / strain:

Chinese hamster Ovary (CHO)

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

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
Osmolarity and pH values were not influenced by test substance treatment.
In this study, in the absence of S9 mix, test substance precipitation was macroscopically observed in culture medium 3 – 4 hours after start of treatment at 200.00 µg/mL in the 1st Experiment, and at 50.00 µg/mL and above in the 2nd Experiment. Additionally, in the presence of S9 mix, precipitates were found in culture medium at the end of treatment at 50.00 µg/mL and above in the 2nd Experiment.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxic effects, as indicated by clearly reduced cloning efficiencies of about or below 20% of the respective negative control values were observed in all three experiments in the absence and presence of S9 mix, at least at the highest applied concentrations.
In the 1st Experiment in the absence of metabolic activation only two surviving test groups (6.25 to 12.50 µg/mL) were obtained after scoring the survival rate (CE1). Due to missing the recommendations of the current OECD Guideline 476 which demands the evaluation of at least four concentrations, this experimental part was discontinued.
In detail, without S9 mix, there was a strong decrease in the number of colonies from 25.00 µg/mL (CE1 relative: 2.5%) onward in the 2nd Experiment and from 20.00 µg/mL (CE1 relative: 0.0%) in the 3rd Experiment, each. The cell densities were distinctly reduced.
In addition, with S9 mix, there was a clear decrease in the number of colonies at 80.00 µg/mL (CE1 relative: 0.0%) in the 1st Experiment and from 50.00 µg/mL (CE1 relative: 11.6%) onward in the 2nd Experiment, respectively. The cell densities were distinctly reduced.

CELL MORPHOLOGY:
After 4 hours of treatment the morphology and attachment of the cells treated with at least the highest applied concentration were adversely influenced in all experimental parts scored for gene mutations. This occurred in samples regardless of the presence or absence of metabolic activation.

Table 1: Summary of results - experimental parts without S9 mix

Exp.	Exposure period [h]	Test groups [µg/mL]	S9 mix	Prec.*	Genotoxicity** MFcorr. [per 10^6cells]	Cytotoxicity***
						CE1 [%]	CE2 [%]
1	4	Vehicle control^1	-	n.d.	n.c.^3	100.0	n.c.^3
		6.25	-	-	n.c.^3	99.0	n.c.^3
		12.50	-	-	n.c.^3	93.1	n.c.^3
		25.00	-	-	n.c.^3	5.7	n.c.^3
		50.00	-	-	n.c.^3	0.0	n.c.^3
		100.00	-	-	n.c.^3	0.0	n.c.^3
		200.00	-	+	n.c.^3	0.0	n.c.³
		Positive control^2	-	n.d	n.c.^3	110.2	n.c.^3
2	4	Vehicle control^1	-	n.d.	4.45	100.0	100.0
		0.78	-	-	n.c.^1	105.3	n.c.^1
		1.56	-	-	3.51	114.8	97.5
		3.13	-	-	0.32	116.3.	99.5
		6.25	-	-	2.33	90.8	99.2
		12.50	-	-	1.12	91.5	109.3
		25.00	-	-	0.65	80.6	102.1
		20.00	-	-	n.c.^2	2.5	n.c.^2
		50.00	-	+	n.c.^2	0.0	n.c.^2
		100.00	-	+.	n.c.²	0.0	n.c.²
		Positive control²	-	n.d.	63.37	113.8	95.7
3	4	Vehicle control^1	-	n.d.	2.69	100.0	100.0
		0.63	-	-	n.c.^1	104.6.	n.c.^1
		1.25	-	-	0.73	109.4	92.4
		2.5.0	-	-	1.85	104.3	93.8
		5.00	-	-	0.00	111.5	96.4
		10.00	-	-	0.65	80.6	102.1
		20.00	-	-	n.c.^2	0.0	n.c.^2
		40.00	-	-	n.c.^2	0.0	n.c.^2
		Positive control^2	-	n.d.	145.02	97.8	85.5

* Precipitation in culture medium at the end of exposure period

** Mutant frequency MF_corr.: mutant colonies per 10⁶ cells corrected with the CE₂ value

*** Cloning efficiency related to the respective vehicle control

n.c.¹ Culture was not continued since a minimum of only four analysable concentrations are required

n.c.² Culture was not continued due to strong cytotoxicity

n.c.³ Culture was not continued due to missing the OECD recommendations

n.d. not determined

1 THF 0.5% (v/v) ² EMS 300 µg/ml

 

Table 2: Summary of results - experimental parts with S9 mix

Exp.	Exposure period [h]	Test groups [µg/mL]	S9 mix	Prec.*	Genotoxicity** MFcorr. [per 10^6cells]	Cytotoxicity***
						CE1 [%]	CE2 [%]
1	4	Vehicle control^1	+	n.d.	1.43	100.0	100.0
		2.50	+	-	2.56	109.0	111.3
		5.00	+	-	4.77	89.1	93.4
		10.00	+	-	2.07	88.4	102.9
		20.00	+	-	2.49	118.4	96.3
		40.00	+	-	0.71	56.1	96.3
		80.00	+	-	n.c.²	0.0	n.c.²
		Positive control^2	+	n.d.	433.00	104.9	78.6
2	4	Vehicle control^1	+	n.d	0.96	100.0	100.0
		3.13	+	-	1.02	91.7	94.2
		6.25	+	-	0.77	79.1	^92.7
		12.50	+	-	2.02	96.1	94.2
		25.00	+	-	0.72	98.8	85.3
		50.00	+	-	0.00	11.6	108.7
		100.00	+	-	n.c.²	0.0	n.c.²
		Positive control^2	+	n.d.	293.78	66.3	67.2

* Precipitation in culture medium at the end of exposure period

** Mutant frequency MF_corr.: mutant colonies per 10⁶ cellscorrected with the CE₂ value

*** Cloning efficiency related to the respective vehicle control

n.c.¹ Culture was not continued since a minimum of only four analysable concentrations are required

n.c.² Culture was not continued due to strong cytotoxicity

n.c.³ Culture was not continued due to missing the OECD recommendations

n.d. not determined

1 THF 0.5%(v/v) ² MCA 20 µg/mL

Conclusions:

The test substance is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation.

Executive summary:

The test substance was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro.

After an attachment period of 20 - 24 hours and a treatment period of 4 hours both with and without metabolic activation and 24 hours without metabolic activation, an expression phase of about 6 - 8 days and a selection period of about 1 week followed. The colonies of each test group were fixed with methanol, stained with Giemsa and counted.

The vehicle controls gave mutant frequencies within the range expected for the CHO cell line. Both positive control substances, EMS and DMBA, led to the expected increase in the frequencies of forward mutations.

In this study, in all experiments at least the highest concentrations tested for gene mutations were clearly cytotoxic in the absence and the presence of metabolic activation.

The 1st Experiment in the absence of S9 mix failed the requirements of the current OECD Guideline 476 due to strong cytotoxicity. A repeat experiment was performed.

On the basis from the results of the present study, the test substance did not cause any relevant and dose-dependent increase in the mutant frequencies both without S9 mix and after adding a metabolizing system in three experiments performed independently of each other.

Conclusion: Thus, under the experimental conditions of this study, the test substance is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation.

Reference 2

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Partly limited documentation. Tested only without metabolic activation.

Principles of method if other than guideline:

Comprehensive testing programme on organophosphorus pesticides comprising in-vitro assay and in-vivo test.

GLP compliance:

not specified

Type of assay:

in vitro mammalian cell micronucleus test

Target gene:

Chinese hamster lung cells

Species / strain / cell type:

other: Chinese hamster lung (CHL) cells

Metabolic activation:

without

Test concentrations with justification for top dose:

different concentration levels (not specified)

Vehicle / solvent:

DMSO

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Statistics:

yes

Species / strain:

other: CHL cells

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: tested up to cytotoxicity threshold

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Conclusions:

Interpretation of results: negative.

Executive summary:

The test substance was assessed for its potential to induce chromosome abberations in an in vitro micronucleus assay as part of a comprehensive testing programme on organophosphorus pesticides.

The assay was conducted on Chinese hamster lung (CHL) cells without metabolic activation. CHL cells were incubated for 48 h at 37°C, then 0.1 ml test substance in DMSO was added and incubated with the test substance for 48 h. The highest dose level was selected to result in 50% survival. Negative (solvent) and positive (0.1 ug/ml) control (presumably concurrent) were also tested. 4000 cells per dose group were examined and the number of nuclei per 1000 cells determined.

Only a short summary table on the results was given, which stated that the test substance was negative in the in virto micronucelus assay.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: comparable to standard guideline with acceptable restrictions; TA 102 and E. coli WP2uvrA not tested, partly limited documentation

Principles of method if other than guideline:

- Method according to Haworth, S. et al.: Environ. Mutagen. 5, Suppl. 1, 3-142, 1983.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Target gene:

S. typhimurium

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat and hamster liver S9 mix

Test concentrations with justification for top dose:

0, 89, 285, 854, 2733 and 8200 ug/plate

Vehicle / solvent:

DMSO

Untreated negative controls:

other: not applicable

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: Positive control in the presence of S9-mix for all strains was 2-aminoanthracene. Without S9-mix: TA 98 4-nitro-ophenylenediamine

Evaluation criteria:

A chemical was judged to be mutagenic (+), or weakly mutagcnic (+W), if it produced a reproducible, dose-related increase in his+ revertants over the corresponding solvent controls in replicate trials. A chemical was considered to be questionable (?) if a reproducible increase of his+ revertants did not meet the criteria for either a " + " or " +W," or if only single doses produced an increase in his+ revertants in repeat trials. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or nonmutagenicity.

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

No increase in the number of revertants at any dose level and in any strain.

Conclusions:

Interpretation of results: negative.

Executive summary:

The test substance was assessed for its potential to induce gene mutations in a bacterial reverse mutation assay as part of a comprehensive testing programme within NTP/USA in which 255 substances were investigated.

The assay was done according to Haworth et al., 1983 (Preincubation method) and covered 4 strains of S. typhimurium: TA 1535, TA1537, TA 98 and TA 100. The test was performed in the presence and absence of rat and hamster liver microsomes (from Arochlor 1254-induced male animals). DMSO was used as vehicle and doses of 0, 89, 285, 854, 2733 and 8200 ug/plate were tested. Concurrent positive and negative (solvent) controls were tested.

No mutagenic activity was observed at doses up to 8200 ug/plate in Salmonella strains TA 98, TA 100, TA 1535 and TA 1537 with or without metabolic activation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

An in vivo micronucleus test in mice was negative (Li et al., 1993).

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: comparable with guideline study with acceptable restrictions; partly limited documentation

Principles of method if other than guideline:

Comprehensive testing programme on organophosphorus pesticides.

GLP compliance:

no

Type of assay:

micronucleus assay

Specific details on test material used for the study:

- purity > 98%

Species:

mouse

Strain:

other: series 615 (no further data)

Sex:

male/female

Route of administration:

intraperitoneal

Vehicle:

corn oil

Details on exposure:

Groups of 10 mice (5 males and 5 females) were treated daily by intraperitoneal injection on four consecutive days with doses corresponding to 80, 40, 20 and 10% of the LD50 (4 day-values; test substance in corn oil). 24 hours after the last treatment, the animals were sacrificed, and bone marrow cells were obtained from the sternum. Slides were produced using standard methods. 1000 polychromatic erythrocytes were scored per animal for the incidence of micronuclei. Cyclophosphamide (100 mg/kg bw, administered on two days) was used as positive control. Negative controls received corn oil (solvent).

Duration of treatment / exposure:

4 days

Frequency of treatment:

daily

Remarks:

80% of the LD50

Remarks:

60% of the LD50

Remarks:

40% of the LD50

Remarks:

20% of the LD50

Remarks:

10% of the LD50

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide (100 mg/kg bw, administered on 2 days)

Tissues and cell types examined:

Bone marrow

Details of tissue and slide preparation:

Slides were produced using standard methods

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Conclusions:

Interpretation of results: negative

Executive summary:

The test substance did not induce micronuclei in bone marrow cells of mice. No details given on results of treated animals, negative and positive controls. However, 6 out of the 21 substances tested in this in vivo micronucleus assay yielded positive results, indicating that the test system was functional.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

In vitro genetic toxicity:

The test substance was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro according to OECD TG 476 and GLP (BASF, 2014). Three independent experiments were carried out with and without the addition of liver S9 mix from phenobarbital- andβ-naphthoflavone induced rats (exogenous metabolic activation). According to the results of the present in vitro study, in three experiments performed independently of each other the test substance did not relevantly increase the number of mutant colonies, either without S9 mix or after the addition of a metabolizing system.

Zeiger et al. (1987) reported a negative result of the test substance in a bacterial reverse mutation assay with and without metabolic activation. Concentations used ranged from 0 - 8200 ug/plate. Test strains used were S. typhimurium TA98, TA100, TA1535 and TA 1537. Negative results in the bacterial reverse mutation assay are published for additional strains like S. typhimurium TA97, TA102 and E. coli WP2 (uvrA), CM891, ND160, MR2 -102, but the documentation of the experimental conditions is very limeted (Li et al., 1993).

Ni et al. (1993) reported a negative result when tested in an in vitro mammalian cell micronucleus assay using CHL cells. Concentrations used were not specified (different concentration levels). This test was done without metabolic activation.

Li et al. (1993) reported a negative result when the test substance was tested in an UDS assay in vitro. No further details are available.

In vivo genetic toxicity:

The test substance did not induce micronuclei in bone marrow cells of mice (Ni et al., 1993). Mice were given daily intraperitoneally doses corresponding to 80, 40, 20 and 10% of the LD50.

Endpoint Conclusion: not genotoxic

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for genotoxicity under Regulation (EC) No. 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.