Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Description of key information

not sensitizing; weight of evidence: OECD TG 406 (Buehler test / Guinea pig maximisation test); RL1-2; GLP

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2002-01-29 to 2002-03-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
July 17, 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
July 30, 1996
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test conducted according to guideline is available, which is reliable without 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.
Species:
guinea pig
Strain:
other: Mol:DH (Moellegaard)
Sex:
male
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Induction: 100 % in deionized water
Challenge: 100 % in deionized water
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Induction: 100 % in deionized water
Challenge: 100 % in deionized water
No. of animals per dose:
3 ‐ preliminary study
10 – main study control
20 – main study treatment
Details on study design:
PRELIMINARY STUDY AND DOSE RANGE FINDING
Left flank: 20 % (in deinonized water), 100% (moistened)
Right flank: 4, 20% (in deinonized water)
Topical application 0.5 g of test substance moistened with 0.5 ml deionized water per plaster (4 cm2) for 6h under fully occluded conditions

INDUCTION
100% (non‐irritating)
Topical application 0.5 g of test substance moistened with 0.5 ml deionized water per plaster (4 cm2) for 6h under fully occluded conditions, three times at intervals of one week. Skin grading 24 and 48h following patch removal

CHALLENGE
100% (non‐irritating); 14 days after the last induction exposure, application of the challenge patch on the shaved skin for 6h; skin evaluations 24 and 48h after patch removal

GRADING SYSTEM
Dermal reactions graded for erythema and edema by blind reading according to grading scale:
- No visible change: 0
- Discrete or patchy erythema: 1
- Moderated and confluent erythema: 2
- Intense erythema and swelling: 3
- not assessable: ?
Positive control substance(s):
yes
Remarks:
Alpha-hexylcinnamic aldehyde
Positive control results:
The validity of the test system is confirmed by the periodically conducted positive control test using alpha-hexyl cinnamic aldehyde for the
Bühler test. No unforeseen circumstances were observed which might have affected the quality or integrity of the study.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25% Alpha-Hexylcinnamaldehyde
Remarks on result:
other: no further details

PRELIMINARY STUDY AND DOSE RANGE FINDING

No irritation at any dose. Concentration for induction and challenge: 100%

MAIN STUDY

No skin reaction observed in the control and test animals treated at the test item at 100% in de‐ionized water.

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance "fully saturated TEA-Esterquat" is not a dermal sensitizer in this study.
Executive summary:

In a dermal sensitization study with the fully saturated TEA-Esterquat, (containing approx 15 % IPA) male Moellegaard guinea pigs were tested using the method of Bühler according to OECD guideline 406. Positive control material was alpha-hexylcinnamic aldehyde.

Dermal induction was performed with undiluted test substance. None of the animals of the test or control group showed any positive skin reactions after challenge treatment with the undiluted test substance. The sensitisation rate at 24 h and at 48 h was 0 %.

 

In this study the test substance, “fully saturated TEA-Esterquat”, is considered to be a non-sensitizer.

 

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2004-01-13 to 2004-02-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
July 17, 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
July 30, 1996
GLP compliance:
yes (incl. QA statement)
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test conducted according to guideline is available, which is reliable without 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.
Species:
guinea pig
Strain:
other: Ibm: GOHI; SPF-quality (Himalayan spotted)
Sex:
male
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
25%, three times at intervals of one week, 6 h exposure
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
1%
Day(s)/duration:
14 d after induction, 6 h exposure
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Total of 38 animals
8 – preliminary screen
10 – main test control
20 – main test treatment
Details on study design:
PRELIMINARY STUDY AND DOSE RANGE FINDING
Left and right cranial and caudal flanks: 1. test: 50, 25, 15, 10; 2. test: 5, 3, 1 and 0.3% (w/w) 50% of test item was applied in a 25 mm Hill top chamber with a spatula.
25, 15, 10, 5, 3, 1 and 0.3% of test items were applied at 0.5 ml in a 25 mm Hill top chamber.
Application for 6h under fully occluded conditions. The application sites were assessed for erythema and oedema approx. 24 and 48 h after removal of the patches.

INDUCTION
25% (minimally irritating); left cranial and caudal flanks; topical application of aqueous solution for 6h, three times at intervals of one week. Skin grading 24h following patch
removal

CHALLENGE
1% (non-irritating); left cranial and caudal flanks; 14d after the last induction exposure, application of the challenge patch on the shaved skin for 6h; skin evaluations 24 and 48h after patch removal

GRADING SYSTEM
Dermal reactions graded for erythema, edema and other clinical changes in skin condition according to grading scale:
No visible change: 0
Discrete or patchy erythema: 1
Moderated and confluent erythema: 2
Intense erythema and swelling: 3
Positive control substance(s):
yes
Remarks:
Alpha-Hexylcinnamaldehyde
Positive control results:
Result: 20/20 (24 h reading) and 17/20 (48 h reading) test animals were observed with descrete/patchy to moderate/confluent erythema after the challenge treatment with the highest tested non-irritating concentration of Alpha-Hexylcinnamaldehyde at 5 % in PEG 300. No skin effect was observed in the control group.
Conclusion: In this study, 100 % (24 h reading) of the animals of the test group were observed with skin reactions after challenge treatment performed with the highest tested non-irritating concentration of Alpha-Hexylcinnamaldehyde at 5 % in PEG 300.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1 % in purfified wate
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
1 % in purified water
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
1 % in purified water
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
1 % in purfied water
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
5% alpha-Hexylcinnamal
No. with + reactions:
20
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
5% alpha-Hexylcinnamal
No. with + reactions:
17
Total no. in group:
20

PRELIMINARY STUDY AND DOSE RANGE FINDING

Concentration for induction: 25% (minimally irritating).

Concentration for challenge:1% (highest non‐irritating dose)

SKIN EFFECTS IN INDUCTION

19/20 (first induction), 20/20 (second and third induction): discrete/patchy to moderate/confluent erythema

0/10 control group: no reactions

SKIN EFFECTS IN CHALLENGE

No skin reaction observed in the control and test animals treated with the test item at 1% in purified water.

Interpretation of results:
GHS criteria not met
Conclusions:
In this study, the test substance partially unsaturated TEA-Esterquat is considered to be a non-sensitizer.
Executive summary:

In a dermal sensitization study according to OECD Guideline 406, July, 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 90 % in approx. 10 % isopropanol) diluted in purified water 20 male young adult Himalayan spotted guinea pigs were tested using the Bühler test method.

The sensitization rate at 24 h and at 48 h was 0 %

In this study, the test substance partially unsaturated TEA-Esterquat is considered to be a non-sensitizer. 

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1994-02-01 to 1994-03-17
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
July 17, 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
December 29, 1992
Principles of method if other than guideline:
Minor deviation: timing of second challenge at 3 % no clear.
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test 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.
Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
female
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
12.5%
Day(s)/duration:
3 times at intervals of one week, 6 h exposure
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
1st challenge: 3 %
2nd challenge: 3 % and 1 %
Day(s)/duration:
14 d (1st challenge) and 21 d (2nd challenge) after last induction exposure; 6 h exposure
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
3 – preliminary study (for induction)
5 – dose finding (for challenge)
10 ‐ control
20 – treated
Details on study design:
PRELIMINARY STUDY AND DOSE FINDING
Preliminary (for induction): left flank: 3, 6, 12.5, 25%
Topical application 0.5 g/plaster (4-6 cm2) aqueous solution (‘viscous to pasty’) for 6h under fully occluded conditions.
Result: Minimally irritating concentration at 12.5%

Dose finding (for challenge): 6, 8, 10, 12%
Topical application 0.5 g/plaster (4-6 cm2) aqueous solution for 6h under fully occluded conditions.

INDUCTION
12.5% (minimally irritating), left cranial flank
Topical application 0.5 g/plaster (4-6 cm2) aqueous solution (‘viscous to pasty’) for 6h under fully occluded conditions, three times at intervals of one week. Skin grading 1 and 24h following patch removal after 3rd induction

CHALLENGE
3% (1st challenge, left and right caudal flanks); 1 % (2nd challenge, left and right caudal flanks); 3% (2nd challenge , right caudal flank only)
14 and 21 days after beginning of last induction (3rd), application of the challenge patch on the shaved skin for 6h; skin evaluation 24, 48 and 72h after patch removal

GRADING SYSTEM
Dermal reactions graded for erythema and edema according to grading scale:
- No reaction: 0
- Partial or slight erythema: 1 (add. grade)
- Weak erythema: 1
- Moderated and diffuse erythema and/or edema: 2
- Strong erythema and/or edema: 3
Positive control substance(s):
not specified
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
3 %
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
grande 1 left and/or right flank
Reading:
1st reading
Hours after challenge:
72
Group:
negative control
Dose level:
3 %
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
grade 1 right flank
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
3 %
No. with + reactions:
5
Total no. in group:
20
Clinical observations:
grade 1 right and/or left flank
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
3 %
No. with + reactions:
11
Total no. in group:
20
Clinical observations:
grade 1 right and/or left flank
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
72
Group:
test chemical
Dose level:
3 %
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
grade 1 left flank
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
3 %
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
grade 1
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
3 %
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
grade 1
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
3 %
No. with + reactions:
5
Total no. in group:
20
Clinical observations:
grade 1, only right flank tested
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
3 %
No. with + reactions:
9
Total no. in group:
20
Clinical observations:
grade 1, only right flank tested
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
72
Group:
test chemical
Dose level:
3 %
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
grade 1, only right flank tested
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
10
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
10
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
10
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
1 %
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
grade 1, only left flank
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
72
Group:
test chemical
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Group:
positive control
Remarks on result:
not measured/tested

PRELIMINARY STUDY

- Minimally irritating concentration at 12.5%

DOSE FINDING

- The result of the dose finding study was a significantly increased irritative sensitivity of the animals therefore concentration for challenge: 3%

Evaluation of dermal effects after challenge with 3 %, re-challenge with 3 and 1 % test substance dilutions in dest. water:

Treatment group:

Animal/

hours

challenge 3 %

re-challenge 1 %

re-challenge 3 %

left flank

right flank

left flank

right flank

right flank

24

48

72

24

48

72

24

48

72

24

48

72

24

48

72

19

0

0

0

1

1

0

0

0

0

0

0

0

0

0

0

20

0

0

0

0

0

0

0

0

0

0

0

0

1

1

0

21

0

0

0

0

1

0

0

0

0

0

0

0

1

1#

0

22

0

1

0

0

0

0

0

0

0

0

0

0

0

0

0

23

1

0

0

1

0

0

0

1

0

0

0

0

0

1

1

24

0

0

0

0

0

0

0

0

0

0

0

0

1

0

0

25

0

1

0

0

0

0

0

0

0

0

0

0

1

0#

0

26

0

0

0

0

1

0

0

0

0

0

0

0

0

0

0

27

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

28

1

1

0

0

0

0

0

0

0

0

0

0

0

0

0

29

0

0

0

0

0

0

0

1

0

0

0

0

0

1

1

30

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

31

1

1

0

1

1

0

0

0

0

0

0

0

0

1

0

32

0

0

0

0

0

0

0

0

0

0

0

0

0

1

0

33

0

0

0

0

1

0

0

0

0

0

0

0

0

0

0#

34

0

0

0

0

0

0

0

1

0

0

0

0

0

0

0

35

0

1

0

0

1

0

0

0

0

0

0

0

0

0

0

36

0

1

0

0

1

0

0

0

0

0

0

0

1

1

1

37

1

0

0

1

0

0

0

0

0

0

0

0

0

0

0

38

0

1

0

0

1

1

0

0

0

0

0

0

0

1

0

# scratch

 

Negative control animals:

 

Animal/

hours

challenge 3 %

re-challenge 1 %

re-challenge 3 %

left flank

right flank

left flank

right flank

 

 

 

24

48

72

24

48

72

24

48

72

24

48

72

24

48

72

4

0

0

0

0

0

0

0

0

0

0

0

0

1

1

0

5

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

7

0

1

0

0

1

0

0

0

0

0

0

0

0

0

0

8

0

0

0

0

1

1

0

0

0

0

0

0

0

1

0

9

0

0

0

0

0

0

0

0

0

0

0

0

1

0

0

10

0

0

0

0

0

0

0

0

0

0

0

0

0

1

0

11

0

1

0

0

0

0

0

0

0

0

0

0

0

0#

0

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0#

0

13

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

# scratch

Interpretation of results:
study cannot be used for classification
Remarks:
ambiguous due to inconsistent pattern
Conclusions:
No conclusion in the report. The study results were evaluated to be ambiguous, due to the partly inconsistent pattern and in most cases rapid fading of response at challenge and re-challenge with 3 % test substance concentration in test and control animals.
Executive summary:

In a dermal sensitization study according to OECD Guideline 406, July, 17, 1992 with the “partially unsaturated TEA-Esterquat” (a.i. 77 % in isopropanole) diluted in water 20 female young adult Pirbright-Hartley guinea pigs were tested using the Bühler test method.

For Induction a mild to moderate irritating concentration of 12.5 % was used. Challenge was performed with a 3 % dilution on the left and right flank. Grade 1 reactions were observed in 25, 55 and 5 % of test animals on right/ and or left flank at the 24, 48 and 72 hour reading, respectively. In 30 % of negative control animals as well a grade 1 reaction was observed at the 48 hour reading on left and/or right flank and in 10 % on the right flank at 72 hours. At re-challenge on the right flank with the same concentration grade 1 reaction was observed in 25, 45 and 15 % of test animals at 24, 48 and 72 hours, respectively. 20 and 30 % of control animals showed grade 1 reactions at 24 and 48 hours. 72 hours after application no effects were observed.

Re-challenge with 1 % revealed no reactions in negative control animals and no reactions on right flank of test animals. Grade 1 response was seen in 3/20 animals at the 48 hours reading, on the left flank. No reactions were observed at the 24 and 72 hour readings in test animals.

Considering only reactions with a clean control group and values of right flank as recommended by the OECD guideline 406 the results of the re-challenge with 1 % are negative with a sensitisation rate of 0 %. However, positive reactions have been observed at re-challenge with 1 % on the left flank, but only at one time point and with grade 1 in 3 animals. A partly inconsistent pattern and in most cases rapid fading of response was observed at challenge and re-challenge with 3 % test substance concentration in test and control animals.

In conclusion the study results were evaluated to be ambiguous.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1994-03-22 to 1994-05-13
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
July 17, 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
December 29, 1992
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test 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.
Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
female
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
1st induction: 60 %
2nd induction: 50 %
3rd induction: 25 %
Day(s)/duration:
3 times at interval of one week, 6 h exposure
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
1st challenge: 10 %
2nd challenge: 3 %
Day(s)/duration:
14 day (1st challenge) and 21 days (2nd challenge) after last induction; 6 h exposure
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
3 – preliminary study (for induction)
10 ‐ control
5 – dose finding (for challenge)
20 – treated
Details on study design:
PRELIMINARY STUDY AND DOSE RANGE FINDING
Preliminary (for induction):
Left flank: 6, 12.5, 25, 50%
Right flank: 50, 60, 80, 100%
Appearance: < 6% liquid; 12.5-25% viscous; >50% high viscous
Topical application 0.5 g/plaster (4-6 cm2) aqueous solution (‘viscous to pasty’) for 6h under fully occluded conditions

Dose range (for challenge): 5, 10, 20, 25%
One week prior to challenge in 5 control animals already treated with vehicle.

MAIN STUDY
INDUCTION
1st induction: 60%, left cranial flank
2nd induction: 50%, left cranial flank
3rd induction: 25%, another left skin area behind
(Lowering the concentration by means of experience.)
Control: vehicle was applied to the left cranial flanks
Topical application 0.5 g/plaster (4-6 cm2) aqueous solution (‘viscous to pasty’) for 6h under fully occluded conditions, three times at intervals of one
week. Skin grading 1 and 24h following patch removal for each induction period

CHALLENGE
10% (1st challenge, right and left caudal flanks); 3% (re-challenge; right and left caudal flanks)
14 and 21 days after beginning of last induction, application of the challenge patch on the shaved skin for 6h; skin evaluations 24, 48 and 72h after patch removal

GRADING SYSTEM
Dermal reactions graded for erythema and edema by blind reading according to grading scale:
- No reaction: 0
- Slight erythema: 0+ (add. grade)
- Weak erythema and/or edema: 1
- Moderated and diffuse erythema and/or edema: 2
- Strong erythema and/or edema: 3
Positive control substance(s):
not specified
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
right and left caudal flanks
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
right and left caudal flanks
Reading:
1st reading
Hours after challenge:
72
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
right and left caudal flanks
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
5
Total no. in group:
20
Clinical observations:
animals with positive response on both flanks, irrespective of severity
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
11
Total no. in group:
20
Clinical observations:
animals with positive response on both flanks, irrespective of severity
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
72
Group:
test chemical
Dose level:
10 %
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
animals with positive response on both flanks, irrespective of severity
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
right and left caudal flanks
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
right and left caudal flanks
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
right and left caudal flanks
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
animals with positive response on both flanks, irrespective of severity
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
animals with positive response on both flanks, irrespective of severity
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
72
Group:
test chemical
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
animals with positive response on both flanks, irrespective of severity
Remarks on result:
no indication of skin sensitisation
Group:
positive control
Remarks on result:
not measured/tested

PRELIMINARY STUDY AND DOSE RANGE FINDING

- Minimally irritating concentration at 60%

-10% concentration for challenge (maximum non‐irritating dose)

Evaluation of dermal effects after challenge with 10 % and re-challenge with 3 % test substance dilutions in dest. water:

Treatment group

Animal/

hours

challenge 10 %

re-challenge 3 %

left flank

right flank

left flank

right flank

24

48

72

144

24

48

72

144

24

48

72

24

48

72

19

0+

1

0+

 

1

1

0+

e

0+

0+

0+

0

0

0

20

0

0

0

 

0

0

0

 

0

0

0

0

0

0

21

0

0

0

 

0

0+

0

 

0

0

0

0

0

0

22

1

1

0+

e

0

0+

0

 

0+

0+

0

0

0

0

23

0

0

0

 

0

0

0

 

0

0

0

0

0

0

24

1

1

1

 

0

1

1

e

0

0

0

0

0

0

25

0

0+

0

 

0

0+

0

 

0

0

0

0

0

0

26

0

0+

0

 

0

0+

0

 

0

0

0

0

0

0

27

1

1

0+

 

0+

0+

0

 

0

0

0

0

0

0

28

0

0+

0

 

0+

0

0

 

0

0

0

0

0

0

29

0

0

0

 

0

0

0

 

0

0

0

0

0

0

30

1

1

1

e

0+

0+

0+

 

0

0

0

0

0

0

31

0

0

0

 

0+

0+

0

 

0

0

0

0

0

0

32

1

0+

0

 

0+

0+

0+

 

0

0

0

0

0

0

33

0

0+

0

 

0

0

0

 

0

0

0

0

0

0

34

0

0

0

 

1

1

1

e

0

0

0

1

0+

1

35

0+

1

0+

 

1

1

0+

 

0

0

0

0

0

0

36

0

0

0

 

0

0

0

 

0

0

0

0

0

0

37

0

0+

0

 

0

0+

0

 

0

0

0

0

0

0

38

0

0+

0

 

0+

1

0+

e

0

0

0

0

0

0

 

e = eschar; e =strong eschar

An additional score 0+ for slight erythema was added by the study authors.

 

For evaluation a response was considered positive, when a reaction on both flanks was observed, irrespective of severity. 

 

All readings for negative control group were zero.

Interpretation of results:
GHS criteria not met
Conclusions:
In this study, partially unsaturated TEA-Esterquat (a.i. 90 %) is not a dermal sensitizer.

Executive summary:

In a dermal sensitization study according to OECD Guideline 406, July, 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 90 %) diluted in purified water 20 female young adult Pirbright-Hartley guinea pigs were tested using the Bühler test method.

Dermal induction was performed at concentrations of 60, 50 and 25 %, due to severity of response the concentration has to be decreased.

At challenge with 10 % test substance on the left and right flank, responses of slight erythema (score 0+, added by the study authors) and weak erythema/edema (score 1) were observed. No response was observed in negative control animals. For evaluation a response was considered positive, when a reaction on both flanks was observed, irrespective of severity. The resulting response rates were 25, 55 and 15 % at the 24, 48 and 72 hour readings, respectively. A consistent response over a period of 2 readings in minimum and a response on the contra lateral flank was observed in 35 % (7/20) of test animals. Re-challenge was performed with a 3 % dilution of the test substance on the left and right flank. Only responses on one flank were observed in 3 animals, most of them with a 0+ score. No reactions were observed in control animals. Due to the failure of response on the contra lateral flank the re-challenge was considered to be negative.

In this study, partially unsaturated TEA-Esterquat is not a dermal sensitizer.

.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1992-06-18 to 1992-07-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
May 12, 1981
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 without 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.
Species:
guinea pig
Strain:
other: Pirbright white Bor:DHPW (SPF)
Sex:
male/female
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
intradermal injection: 5 % in aqua ad inject
topical application: 25 % in aqua ad inject
Day(s)/duration:
intradermal: day 0 / epicutaneous: 7 days after intradermal for 48 h
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
10% in aqua ad inject.
Day(s)/duration:
day 21 / 24 h exposure
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 (10 male, 10 female)
Details on study design:
RANGE FINDING TESTS:
- Intradermal injection:
The test article was diluted with aqua ad inject. and Freund's complete adjuvant (FCA; Sigma, 8024 Deisenhofen) to give a final concentration of 5 %.
Two animals were employed for the concentration tested, skin reactions were recorded 48 h after treatment.

- Dermal Application:
The test article was used undiluted. A closed patch exposure was effected by means of an occlusive bandage. Since this maximum concentration proved irritating, lower concentrations (75, 50, 25, 10 %) were tested. Ten animals were employed and skin reactions were recorded 48 h post applicationem.

MAIN STUDY
A. INDUCTION EXPOSURE (day 0)
- 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 50 % (w/w) diluted in aqua ad inject.
2. 0.1 ml test article diluted in aqua ad inject (final concentration: 5 %)
3. 0.1 ml test article diluted in FCA /aqua ad inject (final concentration: 5 %)
-- Control group:
1. 0.1 ml FCA 50 % (w/w) diluted in aqua ad inject
2. 0.1 ml aqua ad inject. (undiluted)
3. 0.1 ml aqua ad inject 50 % (w/w) diluted in FCA

-- Second stage (day 7)
- 7 days after the intradermal injections the dermal application was initiated subsequent to reclipping of the area 24 hours before application of the test article at the highest concentration producing mild to moderate irritation, i.e. (25 % in aqua ad. inject). The test article was spread in a thick layer [to saturation] over a 4 x 5 cm patch (filter paper). 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 (day 21)
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 (10 % in aqua ad iniect.) 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. 24 and 48 h after patch removal, allergic responses were evaluated on a numerical scale according to Draize.

GRADING SYSTEM
Dermal reactions graded for erythema and edema according to grading scale:
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4

No edema: 0
Very slight edema (barely perceptible): 1
Slight edema (edges well defined by raising): 2
Moderate edema (raised approx 1 mm): 3
Severe edema (raised > 1 mm; beyond area of exposure): 4
Positive control substance(s):
yes
Remarks:
2,4 dinitrochlorobenzene (extreme sensitizer) and benzocaine (moderate sensitizer) are tested periodically. The last test with an acceptable level of response to each of these substances was performed in April 1992.
Positive control results:
2,4 dinitrochlorobenzene (extreme sensitizer) and benzocaine (moderate sensitizer) are tested periodically. The last test with an acceptable level of response to each of these substances was performed in April 1992.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Remarks on result:
other: valid, but not further detailed in the study report

RESULTS

Pilot study (range finding):

- Intradermal: No specific findings were observed.

- Dermal:

-- 100, 75 and 50 %: Partly well-defined erythema and slight oedema with induration of the treated areas were observed.

-- 25 %: Slight to well-defined erythema were observed.

-- 10 %: No skin reactions were observed, highest non-irritant concentration.

Main study:

Signs of irritation during induction:

At a concentration of 25 % slight to well-defined erythema and oedema were observed.

Challenge (conc. of test substance: 10 %, left flank) : The sensitization rate at 24 h and at 48 h was 0 %.

No reactions observed in control and test animals at 24 or 48h.

Interpretation of results:
GHS criteria not met
Conclusions:
The test article "partially unsaturated TEA-Esterquat" is considered to be a non-sensitizer in this study.
Executive summary:

In a dermal sensitization study according to OECD Guideline 406, May, 12 1981 with partially unsaturated TEA-Esterquat (a.i. 90 %, in isopropanol 10 %) diluted in water young adult Pirbright white guinea pigs (10 male, 10 female) were tested using the MAXIMIZATION TEST method.

Intradermal induction was performed with a 5 % test substance concentration. After dermal induction with 25 % solution of test substance, slight to well defined erythema and oedema was observed. None of the animals of the test group or control group showed any positive skin reactions after challenge treatment with a non-irritating test article concentration of 10 % in water. The sensitization rate at 24 h and at 48 h was 0 %.

The test article is considered to be a non-sensitizer in this study. 

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1989-11-17 to 1989-12-24
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1981
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test conducted according to guideline is available, which is reliable without 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.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
Induction: 100 %
Challenge: 100 %
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
Induction: 100 %
Challenge: 100 %
No. of animals per dose:
Total of 34 animals
4 – preliminary study
20 – main study treatment
10 – main study control
Details on study design:
PRELIMINARY STUDY
Preliminary (for induction): 25 ‐ 50 ‐ 75 ‐ 100%
Topical application 0.5 ml of test material, 6h occlusive dermal exposure, grading at 24 and 48h

Dose‐finding (for challenge): 75 ‐ 100%
Topical application 0.5 ml of test material, 6h occlusive dermal exposure, grading at 24 and 48h


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 6-hours
- Test groups: undiluted as supplied
- Site: left flank
- Duration: days 7 and 14
- Concentrations: four different
- Topical application 0.5 g/absorbent lint (4‐6 cm2) of test material (white waxy solid – gently warmed) for 6h under fully occluded conditions, three times at intervals of one week. Skin grading 24h following patch removal at each induction

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: on day 28 (14 d after beginning of last induction)
- Test groups: undiluted as supplied
- Site: right flank
- Concentrations: two different
- Application of the challenge patch on shaved skin for 6h; skin evaluations 24 and 48h after patch removal


GRADING SYSTEM USED
Dermal reactions graded for erythema and edema by blind reading according to grading scale:
No reaction: 0
Scattered mild redness: 1
Moderate and diffuse redness: 2
Intense redness and swelling: 3
Positive control substance(s):
yes
Remarks:
2,4-Dinitrochlorobenzene (DNCB)
Positive control results:
The known contact sensitiser, 2,4 -Dinitrochlorobenzene produced a 15/19 sensitisation rate. This was considered to be a satisfactory sensitisation response for this material under the conditions of the test.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
undiluted as supplied
No. with + reactions:
0
Total no. in group:
19
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
undiluted as supplied
No. with + reactions:
0
Total no. in group:
19
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
undiluted as supplied
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
undiluted as supplied
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Group:
positive control
Dose level:
no data
No. with + reactions:
15
Total no. in group:
19

PRELIMINARY STUDY

Result preliminary (for induction): 100% chosen for induction

Result dose‐finding (for challenge): 100% chosen for challenge

MAIN STUDY

No skin responses observed in any of the test or control animals. One animal (No. 1) was found dead at day 14.

POSITIVE CONTROL SUBSTANCE

The known contact sensitiser, 2,4 -Dinitrochlorobenzene produced a 15/19 sensitisation rate. This was considered to be a satisfactory sensitisation response for this material under the conditions of the test.

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance "fully saturated TEA-Esterquat" is not a dermal sensitizer in this study.
Executive summary:

In a dermal sensitization study with the "fully saturated TEA-Esterquat", undiluted test substance young adult female Dunkin-Hartley guinea pigs were tested using the method of Bühler according to OECD guideline 406. Positive control material was 2,4-Dinitrochlorobenzene.

Dermal induction was performed with undiluted test substance. None of the animals of the test or control group showed any positive skin reactions after challenge treatment with the undiluted test substance. The sensitisation rate at 24 h and at 48 h was 0 %.

In this study the test substance, “fully saturated TEA-Esterquat”, is considered to be a non-sensitizer.

 

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1993-10-13 to 1993-11-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
July 17, 1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test conducted according to guideline is available, which is reliable without 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.
Species:
guinea pig
Strain:
other: Pirbright white Bor: DHPW (SPF)
Sex:
male/female
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
25% (1st and 2nd exposure) and 10% (3rd exposure)
Day(s)/duration:
once weekly for a total of three 6-h exposures
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
10 %
Day(s)/duration:
2 weeks after induction, 6 h exposure
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
study group 20 (10 male, 10 female)
control group 10 (5 male, 5 female)
Details on study design:
RANGE FINDING TESTS:
- The test was conducted in two males and two females using Hill-Top Chambers (Hill Top, Cincinnati, USA) secured with rubber dental dam (4D Rubber Co., Ltd.,UK), the test article was applied in concentrations of 100%, 50%, 25% and 10% to discrete areas of clipped skin on the back of the animals (two concentrations per animal). The animals were then immobilized in metal restrainers for 6 h, after which time the patches were removed and the animals returned to their cages. Skin reactions were recorded 24 h and 48 h after patch removal.

MAIN STUDY
A. INDUCTION EXPOSURE
The induction procedure was performed in both the test and the control group. The test article was applied at a concentration of 25% (first and second exposure) and 10% (third exposure) to an area of clipped skin in the left anterior quadrant of the back of each test animal, whilst the control animals were subjected to an exposure with the vehicle at the same site. Method of application and duration of exposure were the same as in the pilot study. The treated skin site of each animal was evaluated 24 h after patch removal to detect dermal irritation. This procedure was repeated on the same site once weekly for a total of three 6-h exposures.

On the basis of the results of the range finding the concentrations of 25% and 10% of the test article were considered to ie suitable for induction exposure and challange exposure, respectively. Because of mmoderate irritation after the second induction exposure the concentration used for the third induction procedure was reduced to 10%.

After the last induction exposure all animals remained untreated for 2 weeks prior to challenge.

B. CHALLENGE EXPOSURE
Both test and control groups were subjected to a challenge exposure with the test article and the vehicle. The test article was applied in a concentration of 10% to a clipped area of skin in the right posterior quadrant of the back and the vehicle was applied in the right anterior quadrant of each animal. Method of application and duration of exposure were the same as in the pilot study. Approximately 21 h after patch removal and 3 h before the first evaluation the skin area was cleared of hair by reclipping. The evaluation of the skin was performed 24 h and 48 h after the end of the exposure period in order to detect possible allergic responses.
Positive control substance(s):
yes
Remarks:
The reaction to the positive control substance 2-mercaptobenzotbiazole is tested periodically. The last test with an acceptable level of response (extreme sensitizer) to this substance at the concentration of 15% in peanut oil was performed in November 19
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Remarks on result:
other: valid, but not further specified in study report

RESULTS

Pilot study (range finding):

- 100 %: moderate to severe erythema (score 3), moderate oedema (score 2)

- 50 %: well-defined and moderate to severe erythema (scores 2 and 3), very slight to slight oedema (scores 1 and 2)

- 25 %: very slight erythema (score 1), no or very slight oedema (scores 0 and 1)

- 10 %: no skin reactions

Main study:

The sensitization rate at 24 h and at 48 h was 0 %.

Interpretation of results:
GHS criteria not met
Conclusions:
In this study, the test substance partially unsaturated TEA-Esterquat is considered to be a non-sensitizer.
Executive summary:

In a dermal sensitization study according to OECD Guideline 406, Juli 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 90%) in water young adult Pirbright white guinea pig (10 male, 10 female) were tested using the BUEHLER method.

The sensitization rate at 24 h and at 48 h was 0 %

In this study, the test substance partially unsaturated TEA-Esterquat is considered to be a non-sensitizer. 

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have similar (eco)toxicological properties because
• they share structural similarities with common functional groups: One quaternised ethanolamine moiety, one to three, mainly two ester groups with a typical UVCB distribution with long-chain fatty acids of natural origin. The molecular structure is almost identical.
• they are manufactured from similar resp. identical precursors (triethanolamine, long-chain fatty acids, dimethyl sulphate) under similar conditions. Therefore common breakdown products via physical and biological processes, which result in structurally similar chemicals are evident
• A constant pattern in the changing of the potency of the properties across the TEA-Esterquats by chain-length and the grade of esterification is not observed, because the fatty acid chain-length distribution is too narrow and similar and the distribution of mono-, di-, and tri-esters is identical. Some variation caused by variation in C=C double bonds may occur and will be discussed at the relevant endpoint.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See general justification for read-across attached to chapter 13 of this IUCLID file.

3. ANALOGUE APPROACH JUSTIFICATION
See general justification for read-across attached to chapter 13 of this IUCLID file.

4. DATA MATRIX
See general justification for read-across attached to chapter 13 of this IUCLID file.
Reason / purpose for cross-reference:
read-across: supporting information
Reason / purpose for cross-reference:
read-across source
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1-10% in several studies
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
1-10% in several studies
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Group:
negative control
Remarks on result:
no indication of skin sensitisation
Group:
positive control
Dose level:
no details available
Remarks on result:
positive indication of skin sensitisation
Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Reliable relevant data are available for partially unsaturated TEA-Esterquat and the structurally related substance oleic acid-based TEA-Esterquat. A justification for read-across is attached to iuclid section 13.


 


Summary


Two Buehler tests with partially unsaturated TEA-Esterquat were negative, one was judged as ambiguous due to an inconsistent response pattern and in most cases rapid fading of responses at challenge and re-challenge in test and control animals. In one further Buehler test the source substance partially unsaturated TEA-Esterquat was identified as weak sensitiser. Furthermore, two Guinea Pig Maximisation Tests, one conducted with partially unsaturated TEA-Esterquat, the other with oleic acid-based TEA-Esterquat are available, which were both negative.


A weight of evidence approach is used for evaluation. There is no indication from the available specifications to link the weak positive findings in the Buehler studies to differences in the degree of saturation of the underlying alkyl chain in partially unsaturated TEA-Esterquat.


The interpretation of results for those studies which were evaluated to be positive for skin sensitisation were complicated through the occurrence of a partly inconsistent pattern and in most cases rapid fading of response. Evaluation of response was hampered due to irritation and the absence of a clean negative control. Considering the studies with clean negative controls, proper performed induction with 100% test substance or mild to moderate irritating concentrations, there was no (0%) evidence for a skin sensitisation potential.


 


Table16: Reliable sensitisation studies with different methods






























Method



fully


saturated  


TEA-EQ



partially unsaturated TEA-EQ



oleic acid-based 


TEA-EQ



Bühler



2 negative


 



2 negative


1 ambiguous        


1 weak sensitizer


 



 



Maximisation


(Magnussen and Kligman)



 



1 negative


 



1 negative


 



Human HRIPT



2 negative studies


with a total of 208 volunteers



 



 



  


 


Studies in detail


Skin sensitisation


In a dermal sensitization study according to OECD Guideline 406, May, 12 1981 with partially unsaturated TEA-Esterquat (a.i. 90 %, in isopropanol 10 %) diluted in water young adult Pirbright white guinea pigs (10 male, 10 female) were tested using the MAXIMIZATION TEST method.


Intradermal induction was performed with a 5 % test substance concentration. After dermal induction with 25 % solution of test substance, slight to well defined erythema and oedema was observed. None of the animals of the test group or control group showed any positive skin reactions after challenge treatment with a non-irritating test article concentration of 10 % in water. The sensitization rate at 24 h and at 48 h was 0 %. The test item is considered to be a non-sensitizer in this study.  


 


In a dermal sensitization study according to OECD Guideline 406, July 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 90%) in water young adult Pirbright white guinea pig (10 male, 10 female) were tested using the BUEHLER method.


The sensitization rate at 24 h and at 48 h was 0 %. In this study, the test substance partially unsaturated TEA-Esterquat is considered to be a non-sensitizer. 


 


In a dermal sensitization study according to OECD Guideline 406, July, 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 90 % in approx. 10 % isopropanol) diluted in purified water 20 male young adult Himalayan spotted guinea pigs were tested using the Bühler test method.


The sensitization rate at 24 h and at 48 h was 0 %. In this study, partially unsaturated TEA-Esterquat is considered to be a non-sensitizer. 


 


In a dermal sensitization study according to OECD Guideline 406, July, 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 77 % in isopropanol) diluted in water 20 female young adult Pirbright-Hartley guinea pigs were tested using the Bühler test method.


For Induction a mild to moderate irritating concentration of 12.5 % was used. Challenge was performed with a 3 % dilution on the left and right flank. Grade 1 reactions were observed in 25, 55 and 5 % of test animals on right/ and or left flank at the 24, 48 and 72 hour reading, respectively. In 30 % of negative control animals as well a grade 1 reaction was observed at the 48 hour reading on left and/or right flank and in 10 % on the right flank at 72 hours. At re-challenge on the right flank with the same concentration grade 1 reaction was observed in 25, 45 and 15 % of test animals at 24, 48 and 72 hours, respectively. 20 and 30 % of control animals showed grade 1 reactions at 24 and 48 hours. 72 hours after application no effects were observed.


Re-challenge with 1 % revealed no reactions in negative control animals and no reactions on right flank of test animals. Grade 1 response was seen in 3/20 animals at the 48 hours reading, on the left flank. No reactions were observed at the 24 and 72 hour readings in test animals.


Considering only reactions with a clean control group and values of right flank as recommended by the OECD guideline 406 the results of the re-challenge with 1 % are negative with a sensitisation rate of 0 %. However, positive reactions have been observed at re-challenge with 1 % on the left flank, but only at one time point and with grade 1 in 3 animals. A partly inconsistent pattern and in most cases rapid fading of response was observed at challenge and re-challenge with 3 % test substance concentration in test and control animals. In conclusion the study results were evaluated to be ambiguous.


 


In a dermal sensitization study according to OECD Guideline 406, July, 17, 1992 with partially unsaturated TEA-Esterquat (a.i. 90 %) diluted in purified water 20 female young adult Pirbright-Hartley guinea pigs were tested using the Bühler test method.


Dermal induction was performed at concentrations of 60, 50 and 25 %, due to severity of response the concentration has to be decreased.


At challenge with 10 % test substance on the left and right flank, responses of slight erythema (score 0+, added by the study authors) and weak erythema/edema (score 1) were observed. No response was observed in negative control animals. For evaluation a response was considered positive, when a reaction on both flanks was observed, irrespective of severity. The resulting response rates were 25, 55 and 15 % at the 24, 48 and 72 hour readings, respectively. A consistent response over a period of 2 readings in minimum and a response on the contra lateral flank was observed in 35 % (7/20) of test animals. Re-challenge was performed with a 3 % dilution of the test substance on the left and right flank. Only responses on one flank were observed in 3 animals, most of them with a 0+ score. No reactions were observed in control animals. Due to the failure of response on the contra lateral flank the re-challenge was considered to be negative.


In conclusion there is evidence for a weak sensitisation potential of partially unsaturated TEA-Esterquat in this study.


 


In a dermal sensitization study with oleic acid-based TEA-Esterquat in bidest. water, young adult male and female Dunkin-Hartley guinea pigs were tested using the method of Magnusson and Kligman according to OECD guideline 406. Positive control material was 2-Mercaptobenzothiazole.


 In this study, the test substance oleic acid-based TEA-Esterquat produced a sensitisation rate of 15 % (3/19) at the first challenge and a sensitisation rate of 0 (0/20) at re-challenge at a concentration of 1 % and is considered to be a non-sensitizer.


 


There was no evidence for a skin sensitisation potential from two Human Repeated Insult Patch Tests involving a total of 208 volunteers.


 


Conclusion:


A weight of evidence approach covering all three TEA-Esterquats is used for evaluation. It is considered suitable to include all three TEA-Esterquats, differing mainly in the degree of saturation of fatty acid moiety, in the weight of evidence approach.


There is no indication from available specifications, to link the weak positive findings in theBühler studies to differences in the degree of saturation of the underlying alkyl chain in the partially unsaturated TEA-Esterquats.Therefore the result of this weight of evidence approach is justified to be valid without restrictions for this whole TEA-Esterquat group.


According to “Guidance on information requirements and chemical safety assessment Chapter R.7.3.4.2” reliable and relevant human data should be used for hazard identification and are even preferable to animal data. Adjuvant-type tests are likely to be more accurate in predicting a probable skin sensitizing potential of a substance in humans than those methods not employing Freund’s Complete Adjuvant and are thus the preferred of guinea pig methods (Council Regulation EC No 440/2008).


Under REACH, the LLNA is the preferred method for new developed skin sensitisation studies. Existing data of good quality deriving from guinea pig tests will be acceptable and will, if providing clear results, preclude the need for further in-vivo testing (Guidance on information requirements and chemical safety assessment Chapter R.7.3.3.1). Therefore no LLNA has been conducted with partially unsaturated TEA-Esterquat.


Considering the reliable studies, one study of the Bühler type was evaluated as weak positive and one of them was considered ambiguous because an inconsistent pattern of response was observed as outlined below. Further 4 Bühler studies and 2 guinea pig maximization tests were negative for skin sensitization as defined by GHS.


The interpretation of results for those studies which were evaluated to be positive for skin sensitisation were complicated through the occurrence of a partly inconsistent pattern and in most cases rapid fading of response. Evaluation of response was hampered due to irritation and the absence of a clean negative control. The studies rated with Klimisch 3 were not included in the evaluation,because no clear conclusion with regard to classification and labelling could be drawn. Considering the studies with clean negative controls, proper performed induction with 100 % test substance or mild to moderate irritating concentrations, there was no (0%) evidence for a skin sensitisation potential of the TEA-esterquats.


None of the 2 studies involving exposures of a total of 208 human volunteers to TEA-Esterquats was considered to induce a skin sensitisation response in humans.


The ability of chemicals to penetrate the human skin is a pre-requisite to cause a skin sensitisation response. Due to the relatively high molecular weight and physico-chemical properties the dermal penetration of TEA-Esterquats can be considered to be very low as outlined in chapter 5.1 toxicokinetics.


In conclusion, there was no evidence that the investigated TEA-Esterquats induced and/or elicited skin sensitisation responses in humans or the more accurate predicting adjuvant-type method in animals. Equivocal results were observed in the Bühler studies.Taking all evidence from available human and animal data together and emphasising the results of human data and data from guinea pig maximization test, the weight of evidence suggests, that TEA-Esterquats do not represent a skin sensitisation hazard and therefore none of the registered TEA- Esterquats need to be classified for skin sensitisation according to GHS Regulation EC No 1272/2008.


 


Similar results were obtained with the source substance MDEA-Esterquat C16-18 and C18 unsatd., results of Human Repeat Insult Patch Tests and supporting data from two Bühler studies with guinea pigs indicate that the substance does not induce skin sensitisation in humans. These data are included into the dossier to demonstrate, that both substances have a similar toxicological profile.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

There is no information available for respiratory sensitisation. Therefore, there is a data gap in this respect. However, the data gap cannot be fulfilled with experimental data, since there is no internationally accepted animal or in vitro model for respiratory sensitisation. In case human data for respiratory sensitisation emerges, this will be taken into account.

Justification for classification or non-classification

Based on the available data, the substance does not need to be classified for skin sensitisation according to regulation (EC) 1272/2008. Thus, no labelling is required.