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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Five studies on genetic toxicity on undecylenic acid were carried out in the years 1988 and 1989. Four in vitro GLP tests were performed according to corresponding OECD guidelines for the ames test, the cytogenicity test, the in vitro mammalian cell gene mutation and the in vitro DNA repair assay. All tests were negative.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989-07-21 till 1998-09-29
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)
Version / remarks:
1981
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test
Species / strain / cell type:
lymphocytes: human
Details on mammalian cell type (if applicable):
- Type and identity of media: HEPES-buffered RPMI, 20% FCS, 50 ug/ml Gentamycin
- Properly maintained: yes
- Phytohaemaglutinine for stimulation (0.1 ml/10 ml culture)
Metabolic activation:
with and without
Metabolic activation system:
S 9 (Arochlor-induced rat liver)
Test concentrations with justification for top dose:
211.25, 325 and 500 µg/ml
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
Migrated to IUCLID6: and methylmethanesulphonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium in suspension;


DURATION
- Preincubation period: 44 h
- Exposure duration: 3 h
- Fixation time (start of exposure up to fixation or harvest of cells): 3 h (methanol and glacial acetic acid)
SPINDLE INHIBITOR (cytogenetic assays): Colchicine 1 ug/ml added after 2 h to arrest cells in metaphase
STAIN (for cytogenetic assays): Gurr`s Giemase R66


NUMBER OF REPLICATIONS: one male and one female donor, control and all 4 doses with and without S9


NUMBER OF CELLS EVALUATED: 200 (min. 160)
Evaluation criteria:
no data
Statistics:
no data
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Conclusions:
Under the experimental conditions, the test substance undecylenic acid was unable to induce chromosome aberrations in human lymphocytes.
Executive summary:

Undecylenic acid was tested in an in vitro cytogenetic assay according to OECD guideline 473 and EU method B10. The test substance, undecylenic acid, was applied to primary human lymphocyte cultures in the presence or absence of a metabolic activating system (rat liver S9). Treatment of cells with the test subtance in the presence or absence of S9 resulted in similar numbers of aberrations compared to negative controls, while cell cultures treated with positive control substances resulted in significant elevation of aberrations. Under the experimental conditions, the test substance undecylenic acid was unable to induce chromosome aberrations in human lymphocytes.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1981
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine operon
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 -mix from Arochlor-induced rats
Test concentrations with justification for top dose:
10, 50, 100, 250, 500 and 750 µg/ml (genotox);
(bacterial toxicity testing up to 5000 µg/ml)
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: and 2-nitrofluorene, sodium azide, 2-aminoanthracene with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: 20 min.
-Substance Exposure and colony growth (selection): 48h


SELECTION AGENT (mutation assays): histidine-depletion


NUMBER OF REPLICATIONS: 2 studies, each triplicate


DETERMINATION OF CYTOTOXICITY
- Method: counting of formed colonies

Evaluation criteria:
no data
Statistics:
no data
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
not precised
Vehicle controls validity:
not specified
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Conclusions:
In conclusion, undecylenic acid was under the experimental conditions not genotoxic in the AMES test.
Executive summary:

The potential genotoxicity of undecylenci acid was tested by AMES test according to OECD guideline 471 and EU method B.13/14 in five tester strains of salmonella typhimurium. Bacterial cultures were treated with the test substance in the presence and absence of a metabolically activating system (rat liver S-9). Whether in the presence or absence of metabolic activation, no increase was observed in the number of His+-revertant colonies/plate at any of the dose levels tested an any tester strain. In conclusion, undecylenic acid was under the experimental conditions not genotoxic in the AMES test.

Endpoint:
in vitro DNA damage and/or repair study
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 482 (Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells In Vitro)
Version / remarks:
1986
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.18 (DNA Damage and Repair - Unscheduled DNA Synthesis - Mammalian Cells In Vitro)
Deviations:
no
GLP compliance:
yes
Type of assay:
DNA damage and repair assay, unscheduled DNA synthesis in mammalian cells in vitro
Species / strain / cell type:
hepatocytes: rat
Details on mammalian cell type (if applicable):
no
Metabolic activation:
without
Test concentrations with justification for top dose:
test no1 : 1, 5, 10, 25, 50 and 100 µg/ml
test no2 : 10, 25, 50 and 100 µg/ml
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Remarks:
untreated
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
yes
Remarks:
pyren (0.1 mM)
Positive controls:
yes
Positive control substance:
other: 2-aminofluoren (0.01 and 0.05 mM)
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: no data
- Exposure duration: 18-20 hours

NUMBER OF REPLICATIONS: 3 cell cultures

NUMBER OF CELLS EVALUATED: 20 cells/slide

DETERMINATION OF CYTOTOXICITY : yes in preliminary study

OTHER EXAMINATIONS:
- Determination of polyploidy: no data
- Determination of endoreplication: no data
Evaluation criteria:
The result is considered positive when the average nuclear grains in the treated cultures is significantly different from controls and when number of grains in the nucleus subtracted from the number of grains obtained in the cytoplasm was higher than 3.
Statistics:
The test results of DNA repair were quantified by determining the average grain nuclear and cytoplasmic grains, with the standard error in each case.
Species / strain:
lymphocytes: rats
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 100 µg/ml and more
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
See tables

Results of test no1 :

Number of grains per cell

Cytoplasmic grains per cell

Batch no.0 : negative control (untreated)

15+/-0.4

20+/-0.3

Batch no.1 : solvent control (DMSO)

14+/-0.3

20+/-0.4

Batch no.2 : true negative control (pyren)

16+/-0.5

23+/-0.3

Batch no.3 : positive control (2AF, 0.1nM/ml)

82+/-1.9

15+/-0.3

Batch no.4 : positive control (2AF, 0.5nM/ml)

97+/-2.3

13+/-0.3

Batch no.5 : undecylenic acid (100 µg/ml)

14+/-0.4

18+/-0.5

Batch no.6 : undecylenic acid (50 µg/ml)

13+/-0.4

19+/-0.5

Batch no.7 : undecylenic acid (25 µg/ml)

14+/-0.5

21+/-0.5

N=60

Results of test no2 :

Number of grains per cell

Cytoplasmic grains per cell

Batch no.0 : negative control (untreated)

19+/-0.5

23+/-0.4

Batch no.1 : solvent control (DMSO)

20+/-0.4

25+/-0.4

Batch no.2 : true negative control (pyren)

17+/-0.5

22+/-0.5

Batch no.3 : positive control (2AF, 0.1nM/ml)

112+/-0.3

24+/-0.6

Batch no.4 : positive control (2AF, 0.5nM/ml)

141+/-1.8

28+/-0.6

Batch no.5 : undecylenic acid (100 µg/ml)

19+/-0.6

24+/-0.4

Batch no.6 : undecylenic acid (50 µg/ml)

17+/-0.4

22+/-0.4

Batch no.7 : undecylenic acid (25 µg/ml)

16+/-0.4

20+/-0.4

N=60

Conclusions:
Under the experimental conditions, no signs of genotoxicity could be found for the test substance undecylenic acid.
Executive summary:

Undecylenic acid was tested in an in vitro DNA-repair test system for potential genotoxicity acoording to OECD guideline 482 / EU method B.18.

Primary rat hepatocytes cell cultures were treated with the test substance. The amount of thymidine-incorporation into DNA was equivalent in control cell cultures and in cell cultures treated with the test substance up to 500 ug/ml, while thymidine-uptake was significantly increased in cell cultures treated with a positive control substance.

Under the experimental conditions, no signs of genotoxicity could be found for the test substance undecylenic acid.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
1984
Qualifier:
according to guideline
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
yes
Type of assay:
mammalian cell gene mutation assay
Target gene:
hypoxanthine-phosphoribosyl-transferase (HPRT)
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
- Type and identity of media: DMEM 10% FCS, Pen/Strep/fungizone
- Properly maintained: yes
-obtained as cryopreserved stock from Institute de Recherches Scientifiques sur le Cancer, Villejuif, France
Metabolic activation:
with and without
Metabolic activation system:
S 9 (from the liver of Arochlor-induced rats)
Test concentrations with justification for top dose:
10, 30, 100, 300, 500, 600 µg/ml
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
and Dimethylnitrosamin with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk


DURATION
- Preincubation period:
- Exposure duration: 3 h
- Expression time (cells in growth medium): 6 days
- Selection time (if incubation with a selection agent): 10 days on 6-thioguanine

STAIN (for cytogenetic assays): Giemsa


NUMBER OF REPLICATIONS: 2 studies (6 and 10 replicates/condition, respectively)



DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency;
Evaluation criteria:
Mutagen if: 3 times mutation frquency of untreated cells + dose-dependent, statistically significant increase in mutation frequency + reproducible in repeat-experiment
Statistics:
no data
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
up to limit of solubility
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
>= 500 µg/ml in the presence of S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: <= 500 µg/ml

Conclusions:
Under the described experimental conditions, the test substance undecylenic acid is not mutagenic in the chinese hamster V79/HPRT gene mutation assay.
Executive summary:

The in vitro mutagenic activity of the test substance undecylenic acid was investigated in the chinese hamster V79/HPRT-test. The test substance was applied to cell cutures at concentrations of 30 -500 ug/ml in the presence or absence of metabolic activation system (S9 -mix). The mutation frequency of treated cell cultures was equal to that of untreated controls, while cell cultures treated with positive controls presented higher mutation frequencies.

Under the described experimental conditions, the test substance undecylenic acid is not mutagenic in the chinese hamster V79/HPRT gene mutation assay.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

A micronucleus test was performed in the mice according to GLP and OECD guideline. All tests gave negative results on genotoxicity of the test item.

Link to relevant study records
Reference
Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1987-09-22 till 1987-12-16
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
1893
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
Remarks:
(bone marrow)
GLP compliance:
yes
Type of assay:
micronucleus assay
Species:
mouse
Strain:
CD-1
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Chalres River, Cléon, France
- Age at study initiation: 7 weeks
- Weight at study initiation: 29g (males) 22g (females)
- Housing: wire mesh bottom stainless steel cages
- Diet / Water (e.g. ad libitum): ad lib.
- Acclimation period:2 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-2
- Humidity (%): 60+/-10
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
10% gum arabic, freshly prepared
Details on exposure:
no
Duration of treatment / exposure:
single
Frequency of treatment:
single treatment on day 1
Post exposure period:
24, 48 and 72 h
Dose / conc.:
1 000 mg/kg bw/day
Dose / conc.:
2 000 mg/kg bw/day
Dose / conc.:
4 000 mg/kg bw/day
No. of animals per sex per dose:
15
Control animals:
yes, concurrent vehicle
Positive control(s):
Cyclophosphamide ip (15 males and 15 females)
Tissues and cell types examined:
bone marrow smears
Details of tissue and slide preparation:
DETAILS OF SLIDE PREPARATION:
both femurs severed, bone marrow flushed out with calf`s serum, cells opbtained by mild centrifugation and spread over slides after mild homogenization; fixed with methanol and stained automatically

Evaluation criteria:
An increase in micronucleated polychromatic cells shows a possible genotoxic effect.
When the ratio PCE / NCE decreases, this represents marrow cytotoxicity.
Statistics:
no
Sex:
male/female
Genotoxicity:
negative
Remarks:
for all doses similar to that of control animals
Toxicity:
yes
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF DEFINITIVE STUDY
- Ratio of PCE/NCE (for Micronucleus assay): similar for control and 24/48h-groups; 72 h not evaluated
- Appropriateness of dose levels and route: 3 cases of mortality (high dose, only males, 24/48h)
Conclusions:
Under the experimental conditions, undecylenic acid did not reveal any in vivo genotoxic activity in the micronucleus test in mice.
Executive summary:

The in vivo genotoxic potential of undecylenic acid was assessed in the mouse by micronucleus test according to OECD guideline 474 and EU method B.12. Undecylenic acid was administered once by oral gavage at dose levels of 1,2 and 4 g/kg bw to CD-1 mice. Each group consisted of 15 male and 15 female animals. Cyclophosphamide served as positive control.

No clinical signs were reported throughout the study, but three maled of the highest dose group were found dead 24 or 72 h after treatment. The PCE/NCE ratio obtained in all treated animals (24 and 48h after treatment) was similar to that of the control animals, and evaluation of the samples obtained 72 h after treatment was not considered necessary.

Under the experimental conditions, undecylenic acid did not reveal any in vivo genotoxic activity in the micronucleaus test in mice.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Justification for classification or non-classification

Since all genotoxicity tests gave negative results, no classification of undecylenic acid is warranted for genotoxicity properties according to EU regulation (EC) No 1272/2008 (CLP).