1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-(C12-14(even numbered) acyl) derivs., hydroxides, inner salts

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February and March 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

No data on positive control.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

12. May 1981

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A valid Guinea pig maximization test conducted according to guideline with acceptable restrictions is available, which is reliable with restrictions and adequate for classification and labelling purposes. Potency estimation is not mandatory when existing guideline and GLP conforming data are available, which were conducted before the new annex of the REACH Regulation entered into force. Moreover, no indication for skin sensitisation was observed in this study, thus, no dose response information is needed. For this reason and for reasons of animal welfare no additional LLNA was conducted.

Test material

Test material information

Constituent 1

Reference substance name:

1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-(C8-18(even numbered) and C18 unsaturated acyl) derivs., hydroxides, inner salts

EC Number:

931-333-8

Cas Number:

147170-44-3

Molecular formula:

not applicable

IUPAC Name:

1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-(C8-18(even numbered) and C18 unsaturated acyl) derivs., hydroxides, inner salts

Constituent 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

Test material form:

solid - liquid: aqueous solution

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Pirbright white, bor: DHPW (SPF)

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: no data
- Weight at study initiation: 258 - 348 g
- Housing: collective housing up to a maximum of 5 animals per cage (Macrolon type IV)
- Diet: ad libitum, standard laboratory guinea pig diet Ssniff-G
- Water: ad libitum, drinking water as for human consumption
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 2
- Humidity (%): 50 - 85
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): artificial lighting (120 lux), 7.00 a.m. - 7.00 p.m.

Study design: in vivo (non-LLNA)

Induction

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

Induction:
- intradermal: appropriate concentration (concentration not further specified)
- epicutaneous: undiluted test substance as delivered (30 % a.i.). Because the test article was non-irritating at all tested concentrations, the clipped area was pretreated with 10 % sodium lauryl sulfate (SLS) in petrolatum to create a local irritation.

Adequacy of induction:

non-irritant substance, but skin pre-treated with 10% SDS

Challenge

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

Challenge: 30 % a.i., undiluted test substance as delivered

Adequacy of challenge:

other: undiluted test substance as delivered

No. of animals per dose:

20 test animals (10 female, 10 male)
20 control animals (10 female, 10 male)

Details on study design:

RANGE FINDING TESTS:
- Intradermal Injection:
The test article was diluted with equal volumes of aqua ad inject. and Freund's complete adjuvant (FCA; Sigma, Deisenhofen, Germany) to give a final concentration of 5 %.
Two animals were employed for each concentration tested, skin reactions were recorded 48 h and 6 days after treatment.

- Dermal application:
The liquid was used at the highest concentration which did not produce excessive inflammation (30% a.i., undiluted). A closed patch exposure was effected by means of an occlusive bandage. Since this maximum concentration proved irritating, lower concentrations were tested, two animals were employed for each concentration tested and skin reactions were recorded 48 h post applicationem.

MAIN STUDY
A. INDUCTION EXPOSURE
- First stage -an area of 4 x 6 cm over the shoulders was clipped short with electric clippers and cleaned with 70 % (v/v) ethanol. Three pairs of intradermal injections were then made symmetrically in two rows on either side of the spine:
-- Test group:
1. 0.1 ml FCA alone (diluted 1 : 2 in water)
2. 0.1 ml test article alone (in the appropriate concentration)
3. 0.1 ml test article emulsified in FCA (in the appropriate concentration)
-- Control group:
1. 0.1 ml FCA alone (diluted 1 : 2 in water)
2. 0.1 ml aqua ad inject (undiluted)
3. 0.1 ml aqua ad inject (diluted 1 : 2 with FCA)

-- Second stage - 7 days after the intradermal injections, the same area was clipped and cleaned again. Because the test article was non-irritating at all tested concentrations, the clipped area was pretreated with 10 % sodium lauryl sulfate (SLS) in petrolatum. 24 hrs later, the test article was spread in a thick layer [to saturation] over a 2 x 4 cm patch (gauze). The latter was firmly secured over the previous injection sites by an occlusive dressing for 48 h.
Control animals received a patch loaded with the vehicle alone.

B. CHALLENGE EXPOSURE
Both control and test animals were subjected to a challenge exposure 14 days after the second stage of induction. The challenge test was performed on a 5 x 5 cm clipped and shaved area on each flank. The maximal non-irritating concentration of the test article (30 % a.i. undiluted) was applied to the left flank and the vehicle to the right using the patch technique described. In each case the duration of exposure was 24 h under an occlusive
dressing. If necessary, the skin was cleaned with 70 % ethanol 21 h after removal of the patch. 24 and 48 h after patch removal, responses were evaluated on a numerical scale according to Draize.

Positive control substance(s):

not specified

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

30 % a.i.

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

no indication of skin sensitisation

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

30 % a.i.

No. with + reactions:

4

Total no. in group:

20

Clinical observations:

skin reactions score 1 (scattered mild redness)

Remarks on result:

no indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

30 % a.i.

No. with + reactions:

0

Total no. in group:

20

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

30 % a.i.

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

skin reactions score 1 (scattered mild redness)

Any other information on results incl. tables

Pilot study (range finding):

- Intradermal:

After 48 h: No specific findings were observed at 1, 0.5 and 0.1 % (1 females and 1 male animal).

5 %: Black discoloration (6 mm diameter) of the injection site (1 male)

After 6 days: No specific findings were observed at 0.5 and 0.1 % (1 female and 1 male animals).

1 %: Immoderate erythema (8 mm diameter) (1 female, 1 male)

- Dermal:

After 48 h: no reactions after dermal application and at a concentration of 30 % a.i. (undiluted testsubstance as delivered)

Main study:

Test group: At 48 h 4/20 test animals showed skin reactions score 1 (scattered mild redness).

Control group: At 48 h 5/20 test animals showed skin reactions score 1.

The sensitization rate at 24 h and 48 h: 0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

As incidence and severity of reactions seen in the test group were not higher than responses seen in the negative control group, the calculated sensitisation rate was 0. The test substance Coco AAPB is not a dermal sensitizer in this study.

Executive summary:

In a dermal sensitization study with Coco AAPB (30 % a.i.) Pirbright white guinea pig (10 male, 10 female) were tested using the Maximisation Test method according to OECD Guideline 406, May 12, 1981.

The analytical purity of the test item is not stated in study report, according to producer information test material is Coco AAPB and has 30 % a.i. , impurities relevant to sensitisation and referred to a.i. are max 1.7% Alkylamidopropylamine and 33 ppm DMPA (3-dimethylaminopropylamine).

Responses to the challenge procedure were evaluated 24 and 48 h after the end of the exposure period.

There were no skin reactions at the first reading. At the second reading 4/20 test animals and 5/20 control animals showed skin reactions score 1 (scattered mild redness).

As Incidence and severity of reactions seen in the test group were not higher than responses seen in the negative control group, the calculated sensitisation rate was 0.

In this study, Coco AAPB is not a dermal sensitizer.