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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Remarks:
used to justify read-across for other endpoints
Adequacy of study:
supporting study
Study period:
1986-02-04 to 1986-03-29
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report date:
1986

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
Concentration used for induction was not the highest to cause mild irritation as postulated by the OECD guideline 406.
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler 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

Constituent 1
Reference substance name:
Ethanaminium, 2-hydroxy-N-(2-hydroxyethyl)-N,N-dimethyl-, esters with C16-18 and C18-unsatd. fatty acids, chlorides
Cas Number:
1079184-43-2
Molecular formula:
n.a. (UVCB)
IUPAC Name:
Ethanaminium, 2-hydroxy-N-(2-hydroxyethyl)-N,N-dimethyl-, esters with C16-18 and C18-unsatd. fatty acids, chlorides
Constituent 2
Chemical structure
Reference substance name:
Propan-2-ol
EC Number:
200-661-7
EC Name:
Propan-2-ol
Cas Number:
67-63-0
Molecular formula:
C3H8O
IUPAC Name:
propan-2-ol
Test material form:
solid
Specific details on test material used for the study:
- Diethylester dimethyl ammonium chloride
- Physical state: solid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Lippische Versuchstierzucht Hagemann GmbH & Co.
- Age at study initiation: Not available
- Weight at study initiation: 340 grams (estimated)
- Housing: 5 animals in macrolon plastic cages IV (20 cm high, 33 cm width, 55 cm length)
- Diet (e.g. ad libitum): ad libitum Ssniff -G
- Water (e.g. ad libitum):ad libitum,Aqua fontana as for human consumption
- Acclimation period: at least 4 days instead of 5 days recommended by the OECD guideline 406


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 2 °C
- Humidity (%): 50-85 instead of 30 - 70 % recommended by the OECD guideline 406.
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 12 hours daily,light-dark cycle


IN-LIFE DATES: From: 1986-01-13 To: 1986-03-29

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
other: 96% ethanol was used instead of 80 % recommended by the OECD guideline 406
Concentration / amount:
Induction 15 %

One group of 20 test animals were treated with 0.3 ml of 15 % test substance in ethanol while a group of 10 control animals were treated with 0.3 ml of ethanol in Hill Top Chambers for a period of 6 hours weekly for 3 induction exposures.
Day(s)/duration:
6 hours, weekly, 3 induction exposures
Challengeopen allclose all
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
Challenge 15 %
Rechallenge 15 % and 7.5 %
Day(s)/duration:
two weeks after the third induction exposure, 6 h exposure
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
15 % and 7.5 %
Day(s)/duration:
rechallenge 18 days after challenge
No. of animals per dose:
20 animals in the test substance group and 10 animals in the vehicle control group
Details on study design:
RANGE FINDING TESTS:
INDUCTION: The entire back and both sides were clipped one day prior to application. Four animals were exposed for 6 hour period to various concentrations (0.3 ml of each/patch)of the test substance.
Concentrations -20 %, 10 %, 5 %, 2.5 % in ethanol 96 %
Grading: at 24hours and 48 hours
Challenge: a single dermal treatment was performed in 2 guinea pigs using 15% dilution of the test compound in acetone and responses were graded at 24 and 48 hours


MAIN STUDY
A. INDUCTION EXPOSURE
Prior to treatment the left shoulder of each animal was clipped with a small animal clipper. 0,3 ml of the freshly prepared test substance were applied to the Hill Top Chambers and placed on the clipped surface of each animal in the test group and secured with rubber dental dam. Animals were immobilized in metal restrainers for 6 hours. After that time patches were taken off and the test substance was removed with a gentle rinse of warm water before returning the animals to their cages. The procedure was repeated at the same site once a week for the next two weeks.
- No. of exposures: three
- Exposure period: once weekly for three weeks
- Test groups: 20 animals were exposed to 0.3 ml of 15 % in ethanol 96 %
- Control group: 10 animals, procedure not described
- Site: back and both sides of the animal
- Frequency of applications: once a week
- Duration: 6 hours
- Concentrations: 15 % test substance in ethanol 96 %


B. CHALLENGE EXPOSURE
The animals from the test group as well as from the control group were treated following the same patching procedure as described for induction.
- No. of exposures: one 6 hours exposure
- Day(s) of challenge: two weeks after the third induction exposure
- Exposure period: 6 hour under occlusion
- Test groups: 0.3 ml of 15% dilution in acetone
- Control group0.3 ml of 15% dilution in acetone
- Site: patches were applied to a naive skin site
- Concentrations: 15% test substance in acetone
- Evaluation (hr after challenge): 24 and 48 hours


OTHER:
RECHALLENGE:
It was done 18 days after challenge. Test animals and new untreated control animals were treated on naive skin analogously to induction and challenge and grading at 24, 44, 48 hours. Concentrations for rechallgenge 15 % and 7.5 % in acetone.

Grading of all animals was done by positioning them under a four tube fluorescent type light. The control animals were graded before the test animals according to the Scoring system below.
Scores:
No reaction 0
slight patchy erythema ± (0,5)
Slight, but confluent or moderate
Patchy erythema 1
Moderate erythema 2
Severe erythema with or without edema 3
Challenge controls:
Challenge controls was 15 % test substance in acetone. Rechallenge controls were 7.5 % and 15 % test substance in acetone
Positive control substance(s):
no

Results and discussion

Positive control results:
None

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
15% in acetone
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
1/20 animal (grade 1)
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 15% in acetone. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: 1/20 animal (grade 1).
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive response .
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
15% in acetone
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
1/20 animal (grade 1)
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 15% in acetone. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: 1/20 animal (grade 1).
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
15% acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 15% acetone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
7.5% in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 7.5% in acetone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
7.5% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive responses
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 7.5% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive responses.
Reading:
rechallenge
Hours after challenge:
44
Group:
test chemical
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 44.0. Group: test group. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
44
Group:
test chemical
Dose level:
7.5% in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 44.0. Group: test group. Dose level: 7.5% in acetone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
44
Group:
negative control
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 44.0. Group: negative control. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
44
Group:
negative control
Dose level:
7.5% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive responses
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 44.0. Group: negative control. Dose level: 7.5% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive responses.
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
15% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 15% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
7.5% in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no animals with positive response 
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 7.5% in acetone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no animals with positive response .
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
7.5% in acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no animals with positive responses
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 7.5% in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no animals with positive responses.
Group:
positive control
Remarks on result:
not measured/tested

Any other information on results incl. tables

No skin alterations were observed during the induction period with 15 % test substance in ethanol 96%. At challenge, slight erythema was observed in 1/20 test animals. No skin reactions were observed in the control animals. Both test and control animals were rechallenged with 15 % and 7.5% test substance in acetone. No skin responses were observed at rechallenge. Normal weight gains were observed during the study.

It was concluded that the test substance is not a sensitizer.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
MDEA-Esterquat C16-18 and C18 unsatd. did not induce sensitization in the guinea pig model.
Executive summary:

In a dermal sensitization study according to OECD guideline 406; 12 May, 1981 with MDEA-Esterquat C16-18 and C18 unsatd.   (80 % a.i) in IPA, Pirbright - Hartley guinea pig (10 male and 10 female) were tested using the method of Bühler. 

No skin alterations were observed during the induction period with 15 % test substance in ethanol 96%. At challenge, slight erythema was observed in 1/20 test animals. No skin reactions were observed in the control animals. Both test and control animals were rechallenged with 15 % and 7.5 % test substance in acetone. No skin responses were observed at rechallenge.

In this study, MDEA-Esterquat C16-18 and C18 unsatd.  is not a dermal sensitizer.