Registration Dossier

Administrative data

Description of key information

The substanceReaction mass of N,N'-ethane-1,2-diylbis(12-hydroxyoctadecanamide), N,N'-hexane-1,6-diylbis(12-hydroxyoctadecanamide) and N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide) consists of three structurally similar constituents:N,N'-hexane-1,6-diylbis(12-hydroxyoctadecanamide) (CAS no. 55349-01-4, EC no. 611-260-5), N,N'-ethane-1,2-diylbis(12-hydroxyoctadecan-1-amide (CAS no. 123-26-2, EC no. 204-613-6) and N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide) (CAS no. 128554-52-9, EC no. 603-282-9). Experimental data on this substance are not available. The individual constituents and mixtures of them and similar amides are considered suitable read-across substances.

The three constituents of the substance being registered are covered by the two read-across substances 12-hydroxyoctadecanoic acid, reaction products with 1,3-benzenedimethanamine and hexamethylenediamineandN,N'-ethane-1,2-diylbis(12-hydroxyoctadecan-1-amide,which are respectively shown in guideline studies to be not skin sensitising.

The multi-constituent read-across substance 12-hydroxyoctadecanoic acid, reaction products with 1,3-benzenedimethanamine and hexamethylenediamine (CAS no. 220926-97-6, EC no.432-840-2, EC index no.616-201-00-7) includes two constituents of the substance being registered: N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide) and N,N'-hexane-1,6-diylbis(12-hydroxyoctadecanamide). In the provided guideline study according to Annex V using Dunkin-Hartley guinea pigs, this read-across substance is shown to be not skin sensitising. The study reports that dose levels of 5 % and 10 % in Alembicol D of the test substance do not result in skin reactions indicative of sensitisation.

The read-across substance N,N'-ethane-1,2-diylbis(12-hydroxyoctadecan-1-amide) comprises the third constituent of the substance being registered. In the provided guideline study the skin sensitisation was assessed in a mouse local lymph node assay (LLNA) according to OECD Guideline 429 using CBA/Ca mice. Based on the results of this study, this read-across substance is not considered a skin sensitiser.

The aforementioned read-across substances comprise the three constituents of the multi-constituent substance being registered. Therefore, the skin sensitising properties of the substance being registered are expected to be comparable to the properties of the read-across substances.

Consequently, we conclude that the substance being registered does not need to be classified for skin sensitisation. Further testing of the substance is not considered necessary.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04. Apr. 2019 - 12. Jun. 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA/Ca
Sex:
female
Details on test animals and environmental conditions:
Animal Health: The health condition of animals was examined by a veterinarian before initiation of the study. Animals were healthy, without visible signs of disease.
Acclimation: The animals were acclimated in identical conditions as during the experiment for 5 days prior to the start of treatment. The acclimation was according to standard operation procedures.
Housing Condition: The animals were housed in IVC polycarbonate cages (5 animals per cage) suspended on stainless steel racks, in a room equipped with central air-conditioning. The room temperature was within the range of 22 ± 3°C, relative humidity was within the range of 50 - 60 %. The light regimen was set to a 12-hour light / 12-hour dark cycle. The sanitation was performed according to standard operation procedures.
Diet: A laboratory food ssniff (ssniff Spezialdiäten GmbH, Germany) was served ad libitum, each day approximately at the same time.
Water: The animals received tap water for human consumption. Supply of drinking water was unlimited. The quality of drinking water is periodically monitored (including microbiological control) and recorded; certificate of analysis is included in raw data.
Bedding: Lignocel S3/4, Lufa - ITL GmbH, Germany
Animals Identification: Each animal was marked with permanent pen on the tail. Each cage was affixed with a cage card containing pertinent animal and study information.

Vehicle:
dimethyl sulphoxide
Concentration:
100 % (w/v) (pre study)
25, 50, 100 % (w/v) (main study)
No. of animals per dose:
2 (pre study)
5 (main study)
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Positive control results:
SI: 5.57/valid and within the historical values of the testing facility
Key result
Parameter:
SI
Value:
1.01
Test group / Remarks:
25 %
Parameter:
SI
Value:
1.22
Test group / Remarks:
50 %
Parameter:
SI
Value:
1.59
Test group / Remarks:
100 %
Parameter:
EC3
Remarks on result:
not determinable
Remarks:
None of the SIs was higher than 3.
Cellular proliferation data / Observations:
In comparison with the control group, a moderate increasing of the pooled lymph node weights was observed at concentration of 100 %. The pooled lymph node weights of treated groups were 0.0350g for 25 % concentration, 0.0355 g for 50 % concentration and 0.0407 g for 100 % concentration of test item. The lymph node weight of control group and positive control group were 0.0370 g and 0.0734 g, respectively. The DPM values for the three treated groups were 1520.44 (25 %), 1830.86 (50 %) and 2393.11 (100 %), respectively. The SI values for the three treated groups were 1.01 (25 %), 1.22 (50 %) and 1.59 (100 %), respectively. The daily clinical observation of the animals did not show visible clinical signs of toxicity. Administration of the test item in used concentrations did not affect the body weight of treated animals. No erythema was observed in both mice in the pre test after test item administration.
Interpretation of results:
GHS criteria not met
Conclusions:
All validity criteria are fulfilled. The study can be considered as valid. The skin sensitizing potential of Bisamid was assessed using the murine local lymph node assay. Based on the results of this study, Bisamid is not considered a skin sensitizer under the condition of this LLNA study.
Executive summary:

The sensitization potential of Bisamid was evaluated using the Local Lymph Node Assay (LLNA). The LLNA has been developed to determine the allergic contact sensitization potential of chemicals. Based on the recommendations of the OECD Guideline 429, the test item was suspended in DMSO (w/v). The positive control (a-Hexylcinnamic aldehyde) (25 %) was dissolved in Acetone/Olive Oil 4:1. The Pre-screen testwas performed using the dose of 100 %. Based on the observations recorded in the Pre-screen tests, the concentration of 100 % was selected as top dose for the main test. Five female mice (CBA/Ca) per group were topically exposed (dorsum of both ears) to the test item at concentrations of 25 %, 50 % and 100 %, to the positive control and to the vehicle only. Lymphocyte proliferation was measured using incorporation of radioactive125I-iododeoxyuridine and 10-5M fluorodeoxyuridine in the draining lymph nodes. The radioactive incorporation was expressed as disintegrations per minute (DPM)/pooled treatment group and compared with DPM value from the vehicle control group and expressed as the Stimulation Index (SI). After application of the test itemat three concentrations (25 %, 50 % and 100 % w/v) the animals did not show visible clinical symptoms of either local irritation or systemic toxicity. In this study the Stimulation Indices (SI) of 1.01, 1.22 and 1.59 were determined with the test item at concentrations of 25 %, 50 %, and 100 % in DMSO, respectively. The EC3 value could not be calculated, since none of the tested concentrations induced a SI greater than the threshold value of 3. The test item Bisamid is not considered a skin sensitizer under the test conditions of this study.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
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:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
Annex V, Maximisation
GLP compliance:
yes
Type of study:
other: Maximisation Test by Magnusson and Kligman
Justification for non-LLNA method:
The provided study was carried out according to EU Method B.6, in which the Maximisation Test by Magnusson and Kligman is described as an applicable adjuvant-type test method.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
not specified
Route:
intradermal
Vehicle:
other: Alembicol D
Concentration / amount:
Concentration of test material and vehicle used at induction:
Intradermal injection: 5% in Alembicol D
Route:
other: Topical application
Vehicle:
other: Alembicol D
Concentration / amount:
Concentration of test material and vehicle used at induction:
Topical application: 70% in Alembicol D
Vehicle:
other: Alembicol D
Concentration / amount:
Concentration of test material and vehicle used for each challenge:
10 and 5% in Alembicol D
No. of animals per dose:
Number of animals in test group: 10
Number of animals in negative control group: 5
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10 %
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10 %. No with. + reactions: 2.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No w ith. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No w ith. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 5.0.

Maximum concentration not causing irritating effects in preliminary test: 10 %

Signs of irritation during induction:

Intradermal injections:

Slight irritation was seen in test animals at sites receiving test substance, 5% w/v in Alembicol D and slight irritation was observed in control animals receiving Alembicol D.

Topical application:

No erythema was observed in test animals following topical application with test substance, 70% in Alembicol D. No erythema was observed in the control animals.

Evidence of sensitisation of each challenge concentration:

At 10 % - 2/10

At 5 % - 0/10

Other observations:

Dryness and sloughing of the skin was observed at 24 and 48 hours in 5/10 and 2/10 animals challenged with 10%, and in 2/10 and 1/10 animals challenged with 5%. No other signs of toxicity or ill health were observed for any test animals.

Interpretation of results:
GHS criteria not met
Conclusions:
At dose levels of 5 % and 10 % in Alembicol D, 12-hydroxyoctadecanoic acid, reaction products with 1,3-benzenedimethanamine and hexamethylenediamine is shown not to be skin sensitising for Dunkin-Hartley guinea pigs.
Executive summary:

The guideline study on skin sensitisation of 12-hydroxyoctadecanoic acid, reaction products with 1,3-benzenedimethanamine and hexamethylenediamine was conducted according to Annex V using Dunkin-Hartley guinea pigs. The study reports that dose levels of 5 % and 10 % in Alembicol D of the test substance do not result in skin reactions indicative of sensitisation. Conclusively, the GHS criteria for skin sensitisation are not met.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The guideline study on skin sensitisation of the read across substance 1,3-bis[12-hydroxy-octadecamide-N-methylene]-benzene (CAS no. 128554-52-9; 911674-82-3, EC no. 423-300-7, EC index no. 616-198-00-2) is disregarded for the assessment of the skin sensitisation potential of the substance being registered. The study was conducted according to EU Method B.6 using guinea pigs. The study reports that a dose level of 50 % in corn-oil of the test substance results in skin reactions. Although irritation by the dose level was excluded, the results donot allow unambiguous conclusionthat the substance is sensitizing and not irritant.

Furthermore, the identity of 1,3-bis[12-hydroxy-octadecamide-N-methylene]-benzene varies from the constituent of the substance being registered, N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide) (CAS no. 128554-52-9, EC no. 603-282-9). The read-across substance is a UVCB substance, so that additional components are part of the substance which might play a role for toxicity.

N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide) is a constituents of the multi-constituent read-across substance 12-hydroxyoctadecanoic acid, reaction products with 1,3-benzenedimethanamine and hexamethylenediamine (CAS no. 220926-97-6, EC no.432-840-2, EC index no.616-201-00-7), which is shown to be not skin sensitising in the provided guideline study.

Therefore, we conclude that the skin sensitising properties of the UVCB substance 1,3-bis[12-hydroxy-octadecamide-N-methylene]-benzene (CAS no. 128554-52-9; 911674-82-3, EC no. 423-300-7, EC index no. 616-198-00-2) are not applicable to the constituent N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide) (CAS no. 128554-52-9, EC no. 603-282-9). Consequently, the study result for this read-across substance is disregarded for the assessment of skin sensitisation.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The substance Reaction mass of N,N'-ethane-1,2-diylbis(12-hydroxyoctadecanamide), N,N'-hexane-1,6-diylbis(12-hydroxyoctadecanamide) and N,N'-[1,3-phenylenebis(methylene)]bis(12-hydroxyoctadecanamide)consists of three structurally similar bisamide constituents. Experimental data on this substance are not available. The individual constituents and mixtures of them and similar amides are considered suitable read-across substances.

Experimental data is available for two read-across substances: 12-hydroxyoctadecanoic acid, reaction products with 1,3-benzenedimethanamine and hexamethylenediamine and N,N'-ethane-1,2-diylbis(12-hydroxyoctadecan-1-amide,which are respectively shown in guideline studies to be not skin sensitising. The constituents of the read-across substances are also the constituents of the substance being registered. Therefore, the skin sensitising properties of the substance being registered are expected to be comparable to the properties of these read-across substances.

Consequently, we conclude that the substance being registered does not need to be classified for skin sensitisation. Further testing of the substance is not considered necessary.