1-Propanaminium, 2-hydroxy-N-(2-hydroxypropyl)-N,N-dimethyl-, esters with C18-unsatd. fatty acids, Me sulfates (salts)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-01-17 to 2011-02-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

(30 May 2008)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

(17 July 1992)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A valid GPMT conducted according to guideline 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, 2-hydroxy-N-(2-hydroxypropyl)-N,N-dimethyl-, esters with fatty acids, C18 unsatd., Me sulfates (salts)

IUPAC Name:

1-Propanaminium, 2-hydroxy-N-(2-hydroxypropyl)-N,N-dimethyl-, esters with fatty acids, C18 unsatd., Me sulfates (salts)

Test material form:

liquid

Specific details on test material used for the study:

- Name of test material: 1-Propanaminium, 2-hydroxy-N-(2-hydroxypropyl)-N,N-dimethyl-, esters with fatty acids, C18 unsatd., Me-sulfates (salts)
- Physical state: liquid
- Analytical purity: 100%

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Dunkin Hartley, HsdPoc:DH

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: no data
- Weight at study initiation: 320.2 ± 14.0 g (test group) and 293.3 ± 21.2 g (control group)
- Housing: in groups of 2 or 3 animals in Makrolon type IV cages
- Diet (e.g. ad libitum): pelleted diet type "3023", Altromin International, Lage, Germany, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 13 days (for main experiment)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C
- Humidity (%): 30-70%
- Air changes (per hr): 8/h
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: sesame oil

Induction 2

Concentration / amount:

Main experiment:
Intradermal: 0.5 % (w/w)
Dermal: 25 % (w/w)
Challenge: 1 % (w/w)

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: sesame oil

Challenge 2

Concentration / amount:

Main experiment:
Intradermal: 0.5 % (w/w)
Dermal: 25 % (w/w)
Challenge: 1 % (w/w)

No. of animals per dose:

10 (test group), 5 (control group)

Details on study design:

RANGE FINDING TESTS:
pilot experiment with 4 animals to determine which concentration of the test substance
- led to slight irritation after intradermal application (determination of the maximum compatible dose); concentrations tested: 5%, 3.5%, 2% and 0.5% (w/w)
- led to slight irritation after dermal application; concentrations tested: 100%, 75%, 50% and 25% (w/w)
- can just be applied dermally without leading to skin irritation (subirritative dose); concentrations tested: 10%, 5%, 1% and 0.5% (w/w)


MAIN STUDY
A. INDUCTION EXPOSURE
Intradermal induction (day 0)
- No. of exposures: 1
- Test groups: test substance
- Control group: vehicle
- Site: below the shoulder blades
- Frequency of applications: 1
- Concentrations:
0.1 mL FCA (mixed at a ratio of 1+1 in vehicle)
0.1 mL 0.5% test substance in sesame oil (or sesame oil in control)
0.1 mL 0.5% test substance (or sesame oil in control) + FCA (mixed at a ratio of 1+1 in sesame oil)

Topical induction (day 7)
- No. of exposures: 1
- Exposure period: 48 h
- Test groups: test substance
- Control group: sesame oil
- Site: below the shoulder blades, same region as for intradermal induction
- Frequency of applications: 1
- Concentrations: 25%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 21
- Exposure period: 24 h
- Test groups: right flank: vehicle; left flank: test substance in Duhring chambers
- Control group: right flank: vehicle; left flank: test substance in Duhring chambers
- Site: flanks
- Concentrations: 1%
- Evaluation (hr after challenge): 24 h, 48 h

Challenge controls:

yes, vehicle only

Positive control substance(s):

yes

Remarks:

α-hexyl cinnamic aldehyde

Results and discussion

Positive control results:

α-hexyl cinnamic aldehyde in sesame oil: sensitisation rate of 40% (4 of 10 animals positive); experiment from 2011-01-24 to 2011-02-18

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

1%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none reported

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none reported.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none reported

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none reported.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none reported

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none reported.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none reported

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none reported.

Results 5

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

α-hexyl cinnamic aldehyde in sesame oil

No. with + reactions:

4

Total no. in group:

10

Any other information on results incl. tables

Pilot experiment:

Animal	Application	Test substance concentration [%]	Scoring
1	Intradermal	5 3.5 2 0.5	3 3 2 1
2	Dermal without Duhring chambers	100 75 50 25	2 2 2 1
3	Dermal with Duhring chambers (challenge)	10 5	2 1
4	Dermal with Duhring chambers (challenge)	1 0.5	0 0

 

 

Main experiment:

Due to the intended activation of the immune system by FCA slight inflammation and subsequent crust formation was observed at the injection sites with FCA in the test group and the control group animals. This common response is not relevant for the grading.

After intradermal induction 10/10 animals of the test group showed discrete or patchy erythema (grade 1); no skin reactions were seen in the control group. After epicutaneous induction 5/10 animals of the test group showed discrete or patchy erythema (grade 1);no skin reactions were seen in 5/10 animals in the test group and in 5/5 animals in the control group.

After challenge no visible changes of the treated skin sites were observed in the test and control group animals 24 and 48 h after patch removal (= grade 0).

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Conclusions:

MDIPA Esterquat C18 unsatd. was not sensitising in this Guinea pig maximisation test.

Executive summary:

In a dermal sensitisation study according to OECD guideline 406 (17 July 1992) and EU method B.6 (30 May 2008) with MDIPA Esterquat C18 unsatd. (100% a.i.) 10 young adult Dunkin Hartley, HsdPoc:DH guinea pigs were tested using the method of method of Magnusson & Kligman (Guinea Pig Maximisation Test). The positive control α-hexyl cinnamic aldehyde produced a sensitisation rate of 40%.

Test concentrations were selected based on the results of a pilot study: intradermal induction 0.5% in sesame oil; epicutaneous induction 25% on sesame oil; challenge 1% in sesame oil. No pretreatment to create local skin irritation was necessary before epicutaneous induction as the selected concentration was slightly irritating.

After challenge no visible changes of the treated skin sites were observed in the test and control group animals 24 and 48 h after patch removal (= grade 0).

The test material produced a response in 0% of animals. According to CLP, EU GHS (Regulation (EC) No 1272/2008), a response of at least 30% of the test animals of an adjuvant type guinea pig test method for skin sensitisation is considered as positive.

MDIPA Esterquat C18 unsatd. is not a dermal sensitiser in this study.