Fatty acids, tallow, compds. with triethanolamine

Endpoint summary

Description of key information

The test material was considered to be a non-sensitiser under the conditions of the in vivo study in accordance with EU criteria.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 December 2015 to 20 January 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

Buehler test

Justification for non-LLNA method:

The LLNA is the study of choice for skin sensitisation. As detailed in the OECD 429 guideline, despite the advantages of the LLNA, it should be recognised that there are certain limitations that may necessitate the use of TG 406. Chemical groups such as metal salts, organometal, unsaturated compounds and surfactants have been known to be linked to false positive. The Substance is considered to be amphiphilic in nature and therefore would likely have surfactant characteristics, which are known to be falsely positive in the LLNA. Therefore a Guinea Pig Study, OECD 406, was selected to decrease the uncertainty of possible falsely positive effects.

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
For the purpose of the study, the test item was prepared immediately prior to dosing in liquid paraffin for the topical applications (pre-test). This vehicle was chosen as it produced the most suitable formulation at the required concentration. Indeed, the preparation of the test item at 50 % in liquid paraffin (w/w) was a yellow solution.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: 2 or 3 weeks old.
- Weight at study initiation: Between 242 g and 268 g.
- Housing: The animals were housed individually or in groups of two in polycarbonate cages. The flooring of the cages was covered with dust-free wood shavings and the top fitted a stainless steel lid containing a feeding device and drinking device of 500 mL.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: Minimum acclimatization period of 5 days under housing and diet conditions identical to those of the test.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C ± 3 °C
- Humidity (%): from 30 % to 70 %
- Air changes (per hr): At least 10 cycles per hour
- Photoperiod (hrs dark / hrs light): Circadian cycle (12 hrs day/ 12 hrs darkness)

Route:

epicutaneous, occlusive

Vehicle:

paraffin oil

Concentration / amount:

100 % in liquid paraffin

Day(s)/duration:

6 hours

Adequacy of induction:

highest technically applicable concentration used

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

100% test item

Day(s)/duration:

6 hours

Adequacy of induction:

highest technically applicable concentration used

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

paraffin oil

Concentration / amount:

100 % in liquid paraffin

Day(s)/duration:

6 hours

Adequacy of challenge:

highest non-irritant concentration

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

100% test item

Day(s)/duration:

6 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Control group: 10 animals
Test material group: 20 animals

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 local applications
- Exposure period: 6 hours under an occlusive dressing (25mm x 50mm non-woven swab of 4-layer patch from MEDISTOCK held in contact with the skin by means of 50 mm wide hypoallergenic micropore™ adhesive tape from 3M and Blenderm™ from 3M).
- Site: The animals were carefully shorn before each test item application on the inter-scapular zone.
- Frequency of applications: Applications were performed on Day 0, Day 7 and Day 14. Animals were then left untreated for 13 days.
- Concentrations: 100 % diluted at 75 %, 50 %, and 25 % (0.5 mL).
- Removal: Washing of the skin after removal of the dressing was done with liquid paraffin.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Exposure period: 6 hours
- Test groups: On the previously shorn dorso-lumbar zone, an application on either side of the spine, under occlusive dressing (25mm x 50mm non-woven swab of 4-layer patch from MEDISTOCK held in contact with the skin by means of 50 mm wide hypoallergenic micropore™ adhesive tape from 3M and Blenderm™ from 3M).
- Control group: On the previously shorn dorso-lumbar zone, an application on either side of the spine, under occlusive dressing (25mm x 50mm non-woven swab of 4-layer patch from MEDISTOCK held in contact with the skin by means of 50 mm wide hypoallergenic micropore™ adhesive tape from 3M and Blenderm™ from 3M).
- Site: On the previously shorn dorso-lumbar zone on either side of the spine.
- Concentrations: 0.5 mL of the undiluted test material on the left flank and 0.5 mL of liquid paraffin in the right flank.
Removal: Washing of the skin after removal of the dressing was done with liquid paraffin.
- Evaluation (hr after challenge): A macroscopic evaluation of the cutaneous reactions (erythema and oedema) was conducted and all the local or systemic reactions were recorded for animals of the control and treated groups. Approximately 24 hours after removal of the occlusive dressing, the cutaneous reactions were observed and graded according to the scales given below.
- Approximately 24 hours later (i.e. 48 hours after removal of the occlusive dressing), a second observation was made.

Grading scale:
0 No visible change
1 Discrete or patchy erythema
2 Moderate and confluent erythema
3 Intense erythema and swelling

OTHER:
The animals were weighed at the beginning of the study, before the second induction and at the end of the study.

Challenge controls:

The animals of the control groups were subject to the same experimental procedure as the treatment groups.

Positive control substance(s):

no

Positive control results:

alpha-Hexylcinnamaldehyde (CAS: 101-86-0) showed sensitisation at 24 and 48 hours.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

100% liquid paraffin

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none observed

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100 % test item

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none observed

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100 % test item

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none observed

Remarks on result:

no indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

100% liquid paraffin

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none observed

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

50%

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

sensitisation

Remarks on result:

positive indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

50%

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

sensitisation

Remarks on result:

positive indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

25%

No. with + reactions:

3

Total no. in group:

20

Clinical observations:

sensitisation

Remarks on result:

positive indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

25%

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

sensitisation

Remarks on result:

positive indication of skin sensitisation

Preliminary study

24 and 48 hours after removal of the patches, no cutaneous reaction was noted at all tested concentrations. In view of these results, the concentration selected was 100 % for the 3 inductions of the main study and the concentration selected was 100 % (MNIC) for the challenge phase.

Main study:

- Induction phase: After the first induction, no cutaneous reaction was noted in the treated group. After the second induction, discrete erythema was noted in seven animals (7 /20) and no cutaneous reaction was noted in the other animals (13/20) of the treated group.

After the third induction, moderate erythema was noted in one animal (1/20), discrete erythema was noted in one animal (1/20), and no cutaneous reaction was noted in the other animals (18/20) of the treated group.

No cutaneous reaction was recorded during the induction phase in the control group.

- Challenge phase: The concentration selected for the induction phase and the challenge was based on the result of the pre-test. Readings were performed 24 and 48 hours after removal of the patches.

In the treated group (treatment concentration of 100 %), no macroscopic cutaneous reactions attributable to allergy were recorded during the examination following the removal of the occlusive dressing (challenge phase).

In the control group (associated with the treatment concentration of 100 %), no cutaneous reaction was observed during the examination following the removal of the occlusive dressing.

In the treated group on the treated area with liquid paraffin, no macroscopic cutaneous reactions attributable to allergy were recorded during the examination following the removal of the occlusive dressing (challenge phase).

No cutaneous reaction was recorded in animals from the control groups after the challenge phase, on the treated area with liquid paraffin (control item).

Weight evolution

No abnormalities and no differences in the body weight between the control and the treated group were observed.

Mortality

No mortality occurred during the main test.

Clinical signs

No abnormal clinical signs related to the administration of the test item were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material was considered to be a non-sensitiser under the conditions of this study in accordance with EU criteria.

Executive summary:

The potential of the test material to act as a sensitiser was investigated in a GLP study in accordance with the standardised guidelines OECD 406, EU Method B.6 and US EPA OPPTS 870.2600.

After induction of 20 guinea-pigs by 3 topical applications with the test item applied as supplied (100 %) under occlusive dressing and a 13-day rest phase, the challenge phase, under occlusive dressing for 6 hours, consisted to a single topical application of the test item at 100 % and of a negative control (liquid paraffin). The concentration selected for the induction phase and the challenge based on the result of a pre-test. Readings were performed 24 and 48 hours after removal of the patches.

In the treated group (treatment concentration of 100 %), no macroscopic cutaneous reactions attributable to allergy was recorded during the examination following the removal of the occlusive dressing (challenge phase). In the control group (associated with the treatment concentration of 100 %), no cutaneous intolerance reaction was observed during the examination following the removal of the occlusive dressing. In the treated group on the treated area with liquid paraffin, no macroscopic cutaneous reactions attributable to allergy was recorded during the examination following the removal of the occlusive dressing (challenge phase). No cutaneous reaction was recorded in animals from the control groups after the challenge phase, on the treated area with liquid paraffin (control item).

In conclusion, the test material was considered to be a non-sensitiser under the conditions of this study in accordance with EU criteria.

Reference 2

Endpoint:

skin sensitisation: in vitro

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Justification for type of information:

In vitro skin sensitisation studies are not required because adequate data from an in vivo skin sensitisation study is available. The in vivo studies are considered to be reliable and were conducted before the amendment of the REACH annex for the considerations in chemico or in vitro skin sensitisation assays.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Skin sensitisation (in vitro)

In vitro skin sensitisation studies are not required because adequate data from an in vivo skin sensitisation study is available. The in vivo studies are considered to be reliable and were conducted before the amendment of the REACH annex for the consideration of in chemico or in vitro skin sensitisation assays.

Skin sensitisation (in vivo)

The potential of the test material to act as a sensitiser was investigated in a GLP study in accordance with the standardised guidelines OECD 406, EU Method B.6 and US EPA OPPTS 870.2600. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

After induction of 20 guinea-pigs by 3 topical applications with the test item applied as supplied (100 %) under occlusive dressing and a 13-day rest phase, the challenge phase, under occlusive dressing for 6 hours, consisted to a single topical application of the test item at 100 % and of a negative control (liquid paraffin). The concentration selected for the induction phase and the challenge based on the result of a pre-test. Readings were performed 24 and 48 hours after removal of the patches. In the treated group (treatment concentration of 100 %), no macroscopic cutaneous reactions attributable to allergy was recorded during the examination following the removal of the occlusive dressing (challenge phase). In the control group (associated with the treatment concentration of 100 %), no cutaneous intolerance reaction was observed during the examination following the removal of the occlusive dressing. In the treated group on the treated area with liquid paraffin, no macroscopic cutaneous reactions attributable to allergy was recorded during the examination following the removal of the occlusive dressing (challenge phase). No cutaneous reaction was recorded in animals from the control groups after the challenge phase, on the treated area with liquid paraffin (control item).

In conclusion, the test material was considered to be a non-sensitiser under the conditions of this study in accordance with EU criteria.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to skin sensitization.