Registration Dossier

Administrative data

Key value for chemical safety assessment

Additional information

Justification for grouping of substances and read-across

The long-chain aliphatic ester (LCAE) category covers mono-esters of a fatty acid and a fatty alcohol. The category contains both mono-constituent and UVCB substances. The fatty acid carbon chain lengths range is C8 - C22 (even and uneven numbered, including saturated, unsaturated, branched and linear chains) esterified with fatty alcohols with chain lengths from C8 - C22 (even and uneven numbered, including saturated, unsaturated, branched and linear) in varying proportions to mono-esters.

The available data allows for an accurate hazard and risk assessment of the category and the category concept is applied for the assessment of environmental fate and environmental and human health hazards. Thus, where applicable, environmental and human health effects are predicted from adequate and reliable data for source substance(s) within the group, by interpolation to the target substances in the group (read-across approach), applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No 1907/2006. In particular, for each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements for adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across.

A detailed justification for the grouping of chemicals and read-across is provided in the technical dossier (see IUCLID Sections 7.1 and 13) and within Chapter 5.1 of the CSR.

 

Overview of genetic toxicity

CAS

Genetic Toxicity in vitro: gene mutation in bacteria

Genetic Toxicity in vitro: cytogenicity in mammalian cells

Genetic Toxicity in vitro: gene mutation in mammalian cells

Genetic toxicity

in vivo

91031-48-0 (b)

Experimental result:
not mutagenic

--

--

--

26399-02-0

--

Experimental result:
not clastogenic

Experimental result:
not mutagenic

--

868839-23-0

Experimental result:
not mutagenic

Experimental result:
not clastogenic

--

--

3687-46-5

RA: CAS 3234-85 -3,

93803-87-3,

95912-88-2

RA: CAS 3687-45 -4,
93803-87-3,
26399-02-0

RA: CAS 3687-45-4,
26399-02-0

RA: CAS 93803-87-3

59231-34-4 (a)

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

36078-10-1

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

95912-86-0

Experimental result:
not mutagenic

RA: CAS 93803-87-3

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

95912-87-1

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

91031-91-3

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

85116-88-7

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

95912-88-2

Experimental result:
not mutagenic

RA: CAS 3234-85-3,

93803-87-3

RA: CAS 3687-45-4,
93803-87-3,
26399-02-0

RA: CAS 3687-45-4,
26399-02-0

RA: CAS 93803-87-3

3234-85-3

Experimental result:
not mutagenic

RA: CAS 93803-87-3

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

22393-85-7

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

101227-09-2

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

97404-33-6

RA: CAS 3234-85-3,

93803-87-3,

95912-88-2

RA: CAS 3687-45-4,
93803-87-3,
26399-02-0

RA: CAS 3687-45-4,
26399-02-0

RA: CAS 93803-87-3

72576-80-8

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

3687-45-4

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

Experimental result:
not clastogenic

Experimental result:
not mutagenic

--

17673-56-2

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

96690-38-9

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

93803-87-3

Experimental result:
not mutagenic

Experimental result:
not clastogenic

RA: CAS 3687-45-4,

26399-02-0

Experimental result:
not clastogenic

17671-27-1

RA: CAS 3234-85-3, 93803-87-3, 95912-88-2

RA: CAS 3687-45-4, 93803-87-3, 26399-02-0

RA: CAS 3687-45-4,

26399-02-0

RA: CAS 93803-87-3

111937-03-2 (c)

Experimental result:
not mutagenic

--

--

--

(a) Category members subject to the REACh Phase-in registration deadline of 31 May 2013 are indicated in bold font.

(b) Substances that are either already registered under REACh or not subject to the REACh Phase-in registration deadline of 31 May 2013 are indicated in normal font.

(c) Surrogate substances are either chemicals forming part of a related category of structurally similar fatty acid esters or precursors/breakdown products of category members (i.e. alcohol and fatty acid moieties). Available data on these substances are used for assessment of (eco)toxicological properties by read-across on the same basis of structural similarity and/or mechanistic reasoning as described below for the present category.

For all category members registered under REACh a full data set for each endpoint is provided. For substances not subject to the current REACh Phase-in registration, lack of data for a given endpoint is indicated by "--".

Discussion

Genetic toxicity endpoint

CAS 26399-02-0

The cytogenetic potential of 2-ethylhexyl oleate (CAS 26399-02-0) was assessed in an in vitro mammalian chromosome aberration test in primary human lymphocytes, performed according to OECD guideline 473 (Buskens, 2010). Duplicate cultures of human lymphocytes were evaluated for chromosome aberrations in the presence and absence of metabolic activation (rat liver S9-mix). In the first experiment, cells were incubated with test substance concentrations of 3, 10 and 33 µg/mL in ethanol for 3 hours with and without metabolic activation. In the second experiment cells were incubated with 3, 10 and 33 µg/mL for 24 hours followed by 24 hours expression time and 48 hours following 48 hours expression time, all without metabolic activation. 33 µg/mL was chosen as maximum concentration due to limited solubility of the test substance. Evaluation of 100 well-spread metaphase cells from each culture for structural chromosomal aberrations revealed no increase in the frequency of chromosome aberrations and polyploid cells at any dose level in comparison to the negative controls. The test material demonstrated only modest cytotoxicity. The vehicle (solvent) and positive controls were shown to be valid. The test material did not induce a statistically significant increase in the frequency of cells with chromosome aberrations with or without metabolic activation. An in vitro mammalian cell gene mutation assay was performed with 2-ethylhexyl oleate according to OECD 476 (Verspeek-Rip, 2010). Two independent experiments (with 3 or 24 hours of exposure) were performed in mouse lymphoma L5178Y cells in the absence and presence of metabolic activation (S9-mix) with test substance concentrations up to 100 μg/mL dissolved in ethanol. Precipitation was seen at 100 µg/mL and higher. The positive and negative controls were valid and within the range of historical control data. No significant increase in mutation frequency occurred in any of the test conditions.

CAS 3687-46-5

The mutagenic potential of decyl oleate (CAS 3687-46-5) was assessed in a bacterial reverse mutation assay (Ames test) (Gloxhuber, 1979). S. typhimurium strain TA 1535, TA 1537, TA 98, TA 100 and TA 1538 were used in the plate incorporation method in concentrations up to 2500 µg/plate, with and without metabolic activation. The test substance did not induce reversions in any of the S. typhimurium strains with or without metabolic activation. No details were given on the validity of the negative and positive controls, or on cytotoxicity. Due to the limited data on methods and individual results, this study was considered insufficient for assessment.

CAS 95912-86-0

The in-vitro genetic toxicity of Fatty acids, C8-10, C12-18 alkyl esters (CAS 95912-86-0) was assessed in a bacterial reverse mutation assay (Ames test) (Banduhn, 1989). The study was performed equivalent to OECD 471, but without the required TA 102 or E. coli strain. The plate incorporation method was applied using S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 at concentrations up to 5000 µg/plate. The test substance did not induce reversions in any of the S. typhimurium strains with or without metabolic activation. Cytotoxicity was observed at 5000 µg/plate, with and without metabolic activation. All the positive controls were valid.

CAS 95912-87-1

A non-guideline bacterial reverse mutation assay (Ames test) was performed with Fatty acids, C16-18, C12-18 alkyl esters (CAS 95912-87-1) by Wallat (1984). Only a summary report with very limited information was available. S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 were exposed at concentrations up to 5000 µg/plate. The test substance did not induce reversions in any of the S. typhimurium strains with or without metabolic activation. There was no data on the cytotoxicity level, positive control results or negative control results.

CAS 95912-88-2

The mutagenic potential of Fatty acids, C16-18, isotridecyl esters (CAS 95912-88-2) was assessed in a bacterial reverse mutation assay (Ames test) (Banduhn, 1989). The study was performed equivalent to OECD 471, but without the required TA 102 or E. coli strain. S. typhimurium strain TA 1535, TA 1537, TA 98, TA 100 and TA 1538 were used in the plate incorporation method, using concentrations up to 5000 µg/plate, with and without metabolic activation. The test substance did not induce reversions in any of the S. typhimurium strains with or without metabolic activation. No cytotoxicity was observed up to the highest dose level tested and the positive controls were valid.

CAS 3234-85-3

The in-vitro genetic toxicity of tetradecanoic acid, tetradecyl ester (CAS 3234-85-3) was assessed in a bacterial reverse mutation assay (Ames test) performed similarly to OECD 471 (Marquardt, 1995). The preincubation method was applied using S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 at concentrations up to 1000 µg/plate. No TA 102 or E. coli strain was used. The test substance did not induce reversions in any of the S. typhimurium strains with or without metabolic activation. All the positive controls were valid. Precipitation was observed at the highest dose level in the range-finding study with TA 98 and TA 100. More than 50% cytotoxicity was observed in the range-finding study with TA 98 from 10 µg/plate (without metabolic activation) and with TA 100 from 1000 µg/plate (without metabolic activation).

CAS 3687-45-4

The potential of oleyl oleate (CAS 3687-45-4) to induce chromosomal aberrations was assessed using Chinese hamster V79 cells, in a study performed according to OECD 473 (Völkner, 1994). The V79-cells were exposed to oleyl oleate at concentrations up to 100 µg/mL, with and without metabolic activation (S9-mix). One experiment with duplicate replications was performed with short-term treatment (4 h) and fixation time 18 and 28 h, without metabolic activation; and with metabolic activation using 18 h treatment time and 18 h fixation time and 28 h treatment time and 28 h fixation time, respectively. The test material did not induce a statistically significant increase in the frequency of cells with chromosome aberrations, with or without metabolic activation. The mitotic indices of the treated cultures without metabolic activation were 83.4-119% and with metabolic activation 91-127.1%, compared with the vehicle control. Precipitation was observed at concentrations from 100 µg/mL, while no cytotoxicity was noted at any concentration. The vehicle and positive controls were valid.

An in vitro mammalian cell gene mutation assay was performed using oleyl oleate, according to OECD 476 (Poth, 1994). Chinese hamster lung fibroblasts (V79) were treated with oleyl oleate at concentrations of up to 100 µg/mL for 4 h both with and without metabolic activation. After an expression time of 7 days in growth medium, cells were incubated for 9 or 12 days with 6 -thioguanine as selection agent for forward mutation at the HPRT locus. Both with and without metabolic activation, no increases in mutant frequency were observed in the initial and in the confirmatory gene mutation assay. There was no evidence of excessive cytotoxicity (i.e., < 10 % relative cloning efficiency) at any of the tested concentrations either in the presence or absence of metabolic activation in any of the experiments performed.

CAS 17673-56-2

A non-guideline bacterial reverse mutation assay (Ames test) was performed with (Z)-octadec-9-enyl (Z)-docos-13-enoate (CAS 17673-56-2) by Wallat (1982). S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 were exposed to the test substance at concentrations up to 2500 µg/plate. The test substance did not induce reversions in any of the S. typhimurium strains with or without metabolic activation. No information regarding cytotoxicity, precipitation or positive control results were reported. Due to the limited data on results, this study was considered insufficient for assessment.

CAS 93803-87-3

The in vitro genetic toxicity of 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) was assessed in a bacterial reverse mutation assay (Ames test) (Verspeek-Rip, 1998), performed according to OECD 471. S. typhimurium strains TA 1535, TA 1537, TA 98 and TA 100, and E. coli WP2 uvr A were exposed to the test substance at concentrations up to 1000 µg/plate. Precipitation was observed in the medium from 1000 µg/plate and above in all strain, with and without metabolic activation. The test substance did not induce reversions in the S. typhimurium strains or E. coli strain, with or without metabolic activation.

An in vitro chromosomal aberration test was performed with 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) according to OECD 473 (Bertens, 1998). Cultured human peripheral lymphocytes were exposed to the test substance at concentrations up to 1000 µg/mL, with and without metabolic activation (S9-mix). In experiment 1, a short-term treatment (3 h) with harvest time 24 and 48 h, was performed without metabolic activation; while the 24 h treatment time with 24 h harvest time and 48 h treatment time with 48 h harvest time, respectively, was done with metabolic activation. In experiment 2, both the 3 h treatment with metabolic activation and the 24 h treatment without metabolic activation had a harvest time of 24 h. The test material did not induce a statistically significant increase in the frequency of cells with chromosome aberrations, with or without metabolic activation. The mitotic indices of the treated cultures without metabolic activation were 81-113% and with metabolic activation 68-108%, compared with the vehicle control. No cytotoxicity was noted at any concentrations, but precipitation was observed at concentrations from 1000 µg/mL and above. The vehicle and positive controls were valid.

An in vivo mammalian erythrocyte micronucleus test was performed according to OECD Guideline 474, using 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) (Bertens, 1998). 5 mice/sex/dose were administered 500, 1000 and 2000 mg/kg bw of the test substance via intraperitoneal injection and sacrificed after 24 hours. An additional control group and 2000 mg/kg bw treatment group were sacrificed after 48 hours. Bone marrow cells from the femur were extracted, and 2 slides per animal were prepared and stained according to the ‘Wright-stain-procedure’. No increase in the frequency of micronucleated polychromatic erythrocytes was observed in the isolated polychromatic erythrocytes. The treatment groups did not show a decrease in the ratio of polychromatic to normochromatic erythrocytes, compared with the vehicle control groups. No toxicity was observed up to and including the limit dose of 2000 mg/kg bw. The positive control substance (cyclophosphamide) induced a statistically significant increase in the number of micronucleated polychromatic erythrocytes and a reduction in the ratio of polychromatic to normochromatic erythrocytes compared with the vehicle controls, showing the positive control was valid.

 

Overall conclusion for genetic toxicity

Several studies assessing the potential genetic toxicity of category members in vitro in bacteria (Ames test) were available (Banduhn, 1989; Banduhn, 1989; Marquardt, 1995; Verspeek-Rip, 1998). In vitro chromosomal aberration tests were performed with 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) and 2-ethylhexyl oleate (CAS 26399-02-0) respectively (Bertens, 1998; Buskens, 2010). One in vitro mammalian cell gene mutation assay was also available with 2-ethylhexyl oleate (CAS 26399-02-0) (Verspeek-Rip, 2010). The in vivo mammalian erythrocyte micronucleus test in bone marrow was performed with 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) (Bertens, 1998). The results of all the tests were negative. The available data on genetic toxicity, both in vitro and in vivo, indicates that the members of the LCAE category do not have genetic toxicity potential.

A detailed reference list is provided in the technical dossier (see IUCLID, section 13) and within the CSR.


Short description of key information:
In none of these studies mutagenicity in bacteria could be observed.
In none of these studies clastogenic effects in mammalian cells could be observed.
In none of these studies mutagenicity in mammalian cells could be observed.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 "General Requirements for Generation of Information on Intrinsic Properties of substances", information on intrinsic properties of substances may be generated by means other than tests e.g. from information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI, "General rules for adaptation of this standard testing regime set out in Annexes VII to X” states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the group concept is applied to the members of the LCAE category, data will be generated from data for reference source substance(s) to avoid unnecessary animal testing. Additionally, once the group concept is applied, substances will be classified and labelled on this basis.

Therefore, based on the group concept, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.