Reaction mass of bis(N-decyl-N,N-dimethyldecan-1-aminium) carbonate and N-decyl-N,N-dimethyldecan-1-aminium hydrogen carbonate

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

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:

other: GLP study according to international guidelines.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Qualifier:

according to guideline

Guideline:

other: FIFRA 40 CFR

GLP compliance:

yes

Type of study:

Buehler test

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: Young adult
- Weight at study initiation: 300 – 500 g
- Housing: wire mesh suspension cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: At least 9 days

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

- Concentration of irritation applications:
Pilot No. 1: 2.5 %, 1.25 %, 0.5 %, 0.25 %, 0.125 %, 0.05 %, 0.025 %, 0.0125 %
Pilot No. 2: 0.125 %, 0.05 %
- Concentration of induction applications: 0.125 % DDACarbonate
- Concentration of challenge applications: 0.05 % DDACarbonate

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

- Concentration of irritation applications:
Pilot No. 1: 2.5 %, 1.25 %, 0.5 %, 0.25 %, 0.125 %, 0.05 %, 0.025 %, 0.0125 %
Pilot No. 2: 0.125 %, 0.05 %
- Concentration of induction applications: 0.125 % DDACarbonate
- Concentration of challenge applications: 0.05 % DDACarbonate

No. of animals per dose:

See table 1 below.

Details on study design:

STUDY DESIGN
The purpose of the study was to determine the potential of DDACarbonate to produce a photallergenic skin reaction in guinea pigs. The study was conducted in three phases. The first and second phases involved contact primary irritation screening with and without irradiation, respectively. The third phase was composed of the induction and challenge procedures and was considered the definitive study. The test material was administered as a 0.125 % w/v formulation in distilled water for induction and as a 0.05 % w/v in distilled water for challenge.

IRRITATION PHASE
The purpose of the irritating phase was to determine the proper level of test material to be used in the induction and challenge phases. A total of four test material concentrations per animal were evaluated such that each animal in a given four-animal pilot group was exposed to the same four concentrations of test material. Eight test material concentrations were evaluated using two sets of pilot animals. The position of the different concentrations of test material on the animals was varied to adjust for possible site-to-site variation in response. The day following exposure, evaluation of the skin site was made (24-hour reading) and then followed by a second evaluation 24 hours later (48-hour reading). Based on the results of these applications, two minimally to mildly irritating concentrations were chosen for further screening in conjunction with irradiation.
- Days of irritation applications: Single application
- Concentration of irritation applications:
Pilot No. 1: 2.5%, 1.25%, 0.5%, 0.25%, 0.125%, 0.05%, 0.025%, 0.0125%
Pilot No. 2: 0.125%, 0.05%
- Vehicle: Distilled water
- Volume applied: 0.3 mL

INDUCTION
Based on the results of the irritation phase, two concentrations were used in the induction phase. The test material was applied to the prepared skin site and covered with an occlusive cover. Four hours later, the cover was removed and those animals not scheduled for irradiation were returned to the cages. Those scheduled for irradiation were irradiated. No grading of skin sites was done during this phase.
- Days of induction applications:
Group 1: 3 times/week (Monday, Wednesday and Friday) for 3 consecutive weeks (9 applications in total)
Group 2, 5 and 6: untreated
- Concentration of induction applications: 0.125 % DDACarbonate
- Vehicle: See Table 2 below.
- Volume applied: 0.3 mL

CHALLENGE PHASE
Based on the results of the irritation phase, one concentration was used in the challenge phase. The day following exposure, evaluation of the skin site was made (24-hour reading) and then followed by a second evaluation 24 hours later (48-hour reading).
- Days of induction and challenge applications: Single treatment 12 days followed last induction exposure (all groups)
- Concentration of challenge applications: 0.05 % DDACarbonate
- Vehicle: See Table 2 below.
- Volume applied: 0.3 mL

POSITIVE CONTROL
- Positive control substance: 3,3‘,4‘,5-Tetrachlorosalicylanilide
- Concentration of positive control in vehicle: See Table 2 below.
IRRADIATION
Those animals that underwent irradiation (Pilot group No. 2, Groups 1 and 3 in the induction phase and Groups 1, 2, 3 and 5 in the challenge phase) were irradiated for 2 hours, within 30 minutes of completion of respective treatment with test material. Irradiation was achieved using a lighting system of twelve 20 watt black light bulbs. Immediately prior and following the irradiation of the animals, radiation levels were measured using ultraviolet A (UVA) and ultraviolet B (UVB) light probes with peak responses at 310 nm and 365 nm, respectively. The UVA probe was covered with a 3 mm thick piece of window to preclude UVB effects.

Challenge controls:

See table 4 below.

Positive control substance(s):

yes

Remarks:

3,3‘,4‘,5-Tetrachlorosalicylanilide (TSCA)

Positive control results:

See tables 3 and 4 below.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.05 % AI induced test material + irradiation

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.05 % AI induced test material + irradiation . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: 10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.05 % AI induced test material + irradiation

No. with + reactions:

8

Total no. in group:

10

Clinical observations:

2/10 No reaction; 8/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.05 % AI induced test material + irradiation . No with. + reactions: 8.0. Total no. in groups: 10.0. Clinical observations: 2/10 No reaction; 8/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.05 % AI naïve test material + irradiation

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.05 % AI naïve test material + irradiation . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: 10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.05 % naïve test material + irradiation

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.05 % naïve test material + irradiation . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: 10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

Induced TCSA + irradiation

No. with + reactions:

9

Total no. in group:

10

Clinical observations:

3/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction), 5/10 Slight, but confluent or moderate, but patchy erythema, 1/10 Moderate erythema

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: Induced TCSA + irradiation . No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: 3/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction), 5/10 Slight, but confluent or moderate, but patchy erythema, 1/10 Moderate erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

Induced TCSA + irradiation

No. with + reactions:

9

Total no. in group:

10

Clinical observations:

5/10 Slight, patchy erythema, 3/10 Slight, but confluent or moderate, but patchy erythema, 1/10 Moderate erythema

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: Induced TCSA + irradiation . No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: 5/10 Slight, patchy erythema, 3/10 Slight, but confluent or moderate, but patchy erythema, 1/10 Moderate erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

Induced TCSA without irradiation

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: Induced TCSA without irradiation . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: 10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

Induced TCSA without irradiation

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: Induced TCSA without irradiation . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: 10/10 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

Naïve TCSA + irradiation

No. with + reactions:

5

Total no. in group:

5

Clinical observations:

1/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction), 4/5 Slight, but confluent or moderate, but patchy erythema (i.e. a slight but definite reaction at the patch site

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: Naïve TCSA + irradiation . No with. + reactions: 5.0. Total no. in groups: 5.0. Clinical observations: 1/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction), 4/5 Slight, but confluent or moderate, but patchy erythema (i.e. a slight but definite reaction at the patch site.

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

Naïve TCSA + irradiation

No. with + reactions:

5

Total no. in group:

5

Clinical observations:

5/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: Naïve TCSA + irradiation . No with. + reactions: 5.0. Total no. in groups: 5.0. Clinical observations: 5/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

Naïve TCSA without irradiation

No. with + reactions:

5

Total no. in group:

5

Clinical observations:

5/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: Naïve TCSA without irradiation . No with. + reactions: 5.0. Total no. in groups: 5.0. Clinical observations: 5/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

Naïve TCSA without irradiation

No. with + reactions:

3

Total no. in group:

5

Clinical observations:

3/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction)

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: Naïve TCSA without irradiation . No with. + reactions: 3.0. Total no. in groups: 5.0. Clinical observations: 3/5 Slight, patchy erythema (i.e. barely perceptible or questionable reaction).

Table 3: Incidence and severity of skin reaction scores following primary irritation phase

Group	Conc % w/v	Incidence of responses										Mean severity scores
		24-hours					48-hours
		Skin reaction scores					Skin reaction scores
		0	±	1	2	3	0	±	1	2	3	24-hr	48-hr
Pilot No. 1	2.5	0	0	0	4	0	0	0	0	4	0	2	2
	1.25	0	0	0	4	0	0	0	0	4	0	2	2
	0.5	0	0	1	3	0	0	0	1	3	0	1.75	1.75
	0.25	0	2	1	1	0	0	0	3	1	0	1	1.25
	0.125	0	4	0	0	0	0	4	0	0	0	0.5	0.5
	0.05	0	4	0	0	0	0	4	0	0	0	0.5	0.5
	0.025	0	4	0	0	0	0	4	0	0	0	0.5	0.5
	0.0125	0	4	0	0	0	1	3	0	0	0	0.5	0.375
Pilot No. 2	0.125	0	2	0	0	0	0	2	0	0	0	0.5	0.5
	0.05	0	2	0	0	0	0	2	0	0	0	0.5	0.5

  

Table 4: Incidence and severity of skin reaction scores following challenge with test material and TSCA

Group	Incidence of Responses										Mean Severity Scores
	24-Hours					48-Hours
	Skin reaction scores					Skin reaction scores
	0	±	1	2	3	0	±	1	2	3	24-hr	48-hr
Primary Challenge
Group No 1 Induced test material + irradiation	0	10	0	0	0	2	8	0	0	0	0.5	0.4
Group No. 2 Naïve test material + irradiation	0	10	0	0	0	0	10	0	0	0	0.5	0.5
Group No. 3 Induced TCSA + irradiation	1	3	5	1	0	1	5	3	1	0	0.9	0.8
Group No. 4 Induced TCSA without irradiation	0	10	0	0	0	0	10	0	0	0	0.5	0.5
Group No. 5 Naïve TCSA + irradiation	0	4	1	0	0	0	5	0	0	0	0.6	0.5
Group No. 6 Naïve TCSA without irradiation	0	5	0	0	0	2	3	0	0	0	0.5	0.3

 

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Repeated cutaneous application of test item in combination with UV irradiation resulted in mild skin irritation but did not produce evidence of skin sensitisation.

Executive summary:

A study was carried out according to OECD Guideline 406 (Skin Sensitisation) and FIFRA 40 CFR. The purpose of the study was to determine the potential of DDACarbonate to produce a photallergenic skin reaction in guinea pigs. The study was conducted in three phases. The first and second phases involved contact primary irritation screening with and without irradiation, respectively. The third phase was composed of the induction and challenge procedures and was considered the definitive study. The test material was administered as a 0.125 % w/v formulation in distilled water for induction and as a 0.05 % w/v in distilled water for challenge.

Following primary challenge, the incidence and severity of responses in the test groups were comparable to the naïve control group, indicating that photosensitisation had not been induced by the test material.

Repeated cutaneous application of test item in combination with UV irradiation resulted in mild skin irritation but did not produce evidence of skin sensitisation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Migrated from Short description of key information:
A study was carried out according to OECD Guideline 406 (Skin Sensitisation) and FIFRA 40 CFR. The purpose of the study was to determine the potential of DDACarbonate to produce a photallergenic skin reaction in guinea pigs. Repeated cutaneous application of test item in combination with UV irradiation resulted in mild skin irritation but did not produce evidence of skin sensitisation.

Justification for selection of skin sensitisation endpoint:
GLP study according to international guidelines.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the information available the substance is not classified and labeled according to Regulation (EC) No 1272/2008 (CLP) and according to Directive 67/548/EEC (DSD).