Alcohols, C12-14, ethoxylated

Endpoint summary

Description of key information

Skin sensitisation (in vivo, GPMT, OECD 406): not sensitising

Conclusion based on data obtained with alcohols, C12-14, ethoxylated (CAS No. 68439-50-9, EC No. 500-213-3) and considering all available data on skin sensitisation in the Alcohol Ethoxylates (AE) category in a Weight-of-Evidence approach.

 

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 Sep - 27 Oct 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

adopted in 1992

Deviations:

yes

Remarks:

10 test and 5 control animals per group were used instead of 20 test and 10 control animals. Since there is a second test on skin sensitiation in guinea pigs available, no additional testing was performed in the present study.

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

Remarks:

The Department of Health of the Government of the United Kingdom

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Test was done before LLNA as first-choice method for in-vivo testing was set into force.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: David Hall Limited, Staffordshire, United Kingdom
- Age at study initiation: 12 weeks
- Weight at study initiation: 329 - 427g
- Housing: singly or in pairs in solid-floor polypropylene cages furnished with woodflakes
- Diet: Guinea Pig FD1 Diet (Special Diet Services Limited, Witham, Essex, United Kingdom), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22
- Humidity (%): 47-68
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

arachis oil

Concentration / amount:

Intradermal Induction: 1%
Topical Induction: 100%

Day(s)/duration:

epicutaneous induction: 48 h

Adequacy of induction:

other: induction doses based on dose-range finding test using concentrations of 1 and 5% for dermal injection and 25, 50, 75 and 100% for epicutaneous induction

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

arachis oil

Concentration / amount:

Topical Challenge: 25 and 50%

Day(s)/duration:

24 h

Adequacy of challenge:

other: dose levels for topical challenge were selected based on the results of a dose-range finding test; the highest concentration was the highest non-irritant concentration

No. of animals per dose:

10 (in test group), 5 (control)

Details on study design:

RANGE FINDING TESTS:
Two guinea pigs were dermally injected with test item concentrations of 1 and 5%. The animal induced with the 1% formulation showed erythema of grade 3 at the 24, 48 and 72 h reading and erythema of grade 2 at the 7 day reading. The animal induced with the 5% formulation showed erythema of grade 3-4 and green necrosis 24 h after induction and erythema of grade 4 with green necrosis findings and eschar formation at the 48 h, 72 h and 7 day reading. Thus, the 1% formulation was chosen for intradermal induction.
In addition, two animals were topically treated with 25, 50, 75 and 100% concentrations for 48 h. One hour after induction, both animals showed erythema of grade 2-3 at all concentration levels. Edema of grade 2 was observed for both animals at 100% and for one animal at 75 and 50%. Edema of grade 1 was noted in one animal at 25% and at 50 and 75% in the second animal. At the 24 h reading, erythema persisted in both animals at the 75% (score 1) and 100% formulation (score 2) and in one animal at the 50% formulation (score 1). One of the animals shiwed edema formation (grade 1) at 100 and 50%, whereas no edema was observed in the second animal.
At the 48 h reading, both animals showed erythema of grade 2 at the 100% formulation test site and erythema of grade 1 at the 75% formulationt test site. In one of the animals, edema was observed (grade 1) at the 100% formulation test site.
Two additional animals were used for a sighting test on topical challenge. One hour after challenge, erythema of grade 2 was noted in both animals at 100 and 75% and erythema of grade 1 was observed at 50 and 25% in both animals. Edema of gradde 1 was noted at 100% in both animals and at 50% in one of the guinea pigs. At the 24 h reading, both animals showed erythema of grade 1 when treated at 50 and 100%. Erythma of score 1 at challenge wirh 100% persisted in one of the animals up to the 48 h time point. At the 24 and 48 h reading, no finding of edema was observed in any animal.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections): 0.1 mL each
Injection 1: 1:1 mixture complete Freund's adjuvant (FCA) and distilled water
Injection 2: 1% test substance in arachis oil BP
Injection 3: 1% test substance in a 1:1 mixture of complete Freund's adjuvant (FCA) and distilled water
Epicutaneous: filter paper patch saturated with undiluted test material
- Control group:
Intradermal (3 pairs of injections):
Injection 1: 1:1 mixture complete Freund's adjuvant (FCA) and distilled water
Injection 2: 100% arachis oil BP
Injection 3: a 50% formulation of arachis oil BP in a 1:1 mixture of complete Freund's adjuvant (FCA) and distilled water
Epicutaneous: filter paper patch saturated with arachis oil BP
- Site: shoulder region
- Frequency of applications: topical induction was performed one week after the intradermal induction
- Duration: 7 days
- Concentrations: intradermal 1%, epicutaneous 100%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 3 weeks after intradermal induction (study Day 21)
- Exposure period: 24 h
- Test groups: test substance and vehicle
- Control group: test substance and vehicle
- Site: left flank (test item) and right flank (vehicle)
- Concentrations: 25 and 50%
- Evaluation (hr after challenge): 48 and 72 h (24 and 48 h after dressing removal)

Challenge controls:

The control group is actually a challenge control

Positive control substance(s):

yes

Remarks:

Historical data on ethyl-4-aminobenzoate, 2,4-dinitrochlorobenzene, neomycin sulphate and 2-mercaptobenzothiazole were provided.

Positive control results:

Historical control data on several positive control chemicals were provided. Challenge with 2-mercaptobenzothiazole (intrademally induced with 10% in arachis oil, topically induced with 50% in acetone and topically challenged with 25 and 50% in acetone : PEG 400 in a 70:30 ratio), exhibited dermal alterations in 70% of the test animals, thus meeting the reliability criteria for the GPMT test (≥ 15% positive reactions).

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

intradermal induction: 1%; epicutaneous induction: 100%; epicutaneous challenge: 25%

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

intradermal induction: 1%; epicutaneous induction: 100%; epicutaneous challenge: 25%

No. with + reactions:

0

Total no. in group:

10

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

challenge: 25%

No. with + reactions:

0

Total no. in group:

5

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

challenge: 25%

No. with + reactions:

0

Total no. in group:

5

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

intradermal induction: 1%; epicutaneous induction: 100%; epicutaneous challenge: 50%

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

intradermal induction: 1%; epicutaneous induction: 100%; epicutaneous challenge: 50%

No. with + reactions:

0

Total no. in group:

10

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

challenge: 50%

No. with + reactions:

0

Total no. in group:

5

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

challenge: 50%

No. with + reactions:

0

Total no. in group:

5

Group:

positive control

Dose level:

intradermal induction: 10%; epicutaneous induction: 50%; epicutaneous challenge: 25 and 50%

Remarks on result:

other: Historical positive control data provided in study report

Local skin reactions:

After intradermal induction, well-defined or moderate to severe erythema was noted at the induction sites of all test group animals at the 24- and 48-h observation. Very slight erythema was noted at the intradermal induction sites of 4 control animals at the 24 and 48 h reading.

After topical induction, very slight to moderate to severe erythema with or without very slight edema was noted at the induction sites of all test group animals at the 1-h observation. Very slight to well-defined erythema with or without very slight to slight edema was noted at the induction sites of seven test group animals at the 24-h observation. Dried blood was noted at the induction sites of six test group animals at the 24 -h observation. Skin reactions prevented the accurate evaluation of erythema and edema at the induction sites of two test group animals at the 24 -h observation. No skin reactions were noted at the treatment sites of control group animals at the 1- and 24-h observations.

After topical challenge, no skin reactions were observed in any control or test item-treated animal at the 24 and 48 -h observation.

Table 1: Skin sensitisation results

Animals Number	skin Reactions (Hours After Removal of Dressing)
	24 h						48 h
	50%			25%			50%			25%
	Er	Oe	Other	Er	Oe	Other	Er	Oe	Other	Er	Oe	Other
1	0	0	-	0	0	-	0	0	-	0	0	-
2	0	0	-	0	0	-	0	0	-	0	0	-
3	0	0	-	0	0	-	0	0	-	0	0	-
4	0	0	-	0	0	-	0	0	-	0	0	-
5	0	0	-	0	0	-	0	0	-	0	0	-
6	0	0	-	0	0	-	0	0	-	0	0	-
7	0	0	-	0	0	-	0	0	-	0	0	-
8	0	0	-	0	0	-	0	0	-	0	0	-
9	0	0	-	0	0	-	0	0	-	0	0	-
10	0	0	-	0	0	-	0	0	-	0	0	-

Er=erythema

Oe=oedema

- = no other reactions noted

Er=erythema

Oe=oedema

- = no other reactions noted

Systemic toxicity: Not reported.

Body weight: All animals gained weight as expected.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Executive summary:

A study was performed to assess the contact sensitisation potencial of the test material in the albino guinea pig. Ten test and five control animals were used for the main study. Based on the results of sighting test, the concentration of the test material for the induction and challenge phases were selected as follows:

Intradermal Induction: 1% w/v in arachis oil BP

Topical Induction: undiluted as supplied

Topical Challenge: 50% and 25% v/v in arachis BP

The test material produced a 0% (0/10) sensitisation rate and was classified as a NON-SENSITISER to guinea pig skin. The test material did not meet the criteria for classification as a sensitiser according to EU labelling regulations. No risk phrase is required.

Reference 2

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

20 Sep- 21 Oct 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

adopted in 1992

Deviations:

yes

Remarks:

no reliablity check reported; control animals were not handled according to guideline (no application of vehicle) during 2nd induction.

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Test was done before LLNA as first-choice method for in-vivo testing was set into force.

Species:

guinea pig

Strain:

other: Pirbright white

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Interfauna, Tuttlingen, Germany
- Females nulliparous and non-pregnant: yes
- Weight at study initiation: 314.6 and 323.1 g (main study) and 397.4 and 400.3 g (sighting study)
- Housing: In groups of 3 in Makrolon type IV cages
- Diet: Altromin Haltungsdiät 3032 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 50 - 60
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

Intradermal induction: 0.1%
Epicutaneous induction: 15%

Day(s)/duration:

epicutaneous induction: 48 h

Adequacy of induction:

other: induction dose levels based on a range-finding test performed, in which intradermal injection at 0.1% caused sufficient irritation; topical induction at 15% caused slight to well-defined skin reactions

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

Challenge: 2.5 and 5%

Day(s)/duration:

24 h

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

20

Details on study design:

RANGE FINDING TESTS:
Three guinea pigs were dermally injected with test item concentrations of 0.1, 0.5, 1.0 and 2.0%. The formulation of 0.1% produced sufficient irritation in the animals, therefore 0.1% was chosen as concentration for the intradermal injection.
In addition, three animals were topically treated with 5, 10 and 15% concentrations for 48 h. The 15% caused slight to well defined skin reactions in all animals, whereas the 10% formulation caused a weak response in 2/3 animals. Thus, 15% was chosen as concentration for topical induction and 2.5 and 5% were chosen as concentrations for topical challenge.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections): 0.1 mL each
Injection 1: 1:1 mixture complete Freund's adjuvant (FCA) and physiological saline
Injection 2: 0.1% test substance in water
Injection 3: 0.2% test substance in a 1:1 mixture of complete Freund's adjuvant (FCA) and physiological saline
Epicutaneous:
- Control group:
Intradermal (3 pairs of injections):
Injection 1: 1:1 mixture complete Freund's adjuvant (FCA) and physiological saline
Injection 2: not performed as water was used as solvent
Injection 3: a 1:1 mixture of complete Freund's adjuvant (FCA) and physiological saline and water
Epicutaneous:
- Site: shoulder region
- Frequency of applications: topical induction was performed one week after the intradermal induction
- Duration: 7 days
- Concentrations: intradermal 0.1%, epicutaneous 15%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 14 days after intradermal induction (study Day 21)
- Exposure period: 24 h
- Test groups: test substance and vehicle
- Control group: test substance and vehicle
- Site: left flank
- Concentrations: 2.5 and 5.0%
- Evaluation (hr after challenge): 48 and 72 h (24 and 48 h after patch removal)

Challenge controls:

The control group is actually a challenge control

Positive control substance(s):

no

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

challenge: 2.5%

No. with + reactions:

0

Total no. in group:

18

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

challenge: 2.5%

No. with + reactions:

0

Total no. in group:

18

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

intradermal induction: 0.1%; epicutaneous induction: 15%; epicutaneous challenge: 2.5%

No. with + reactions:

0

Total no. in group:

19

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

intradermal induction: 0.1%; epicutaneous induction: 15%; epicutaneous challenge: 2.5%

No. with + reactions:

0

Total no. in group:

19

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

challenge: 5%

No. with + reactions:

4

Total no. in group:

18

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

challenge: 5%

No. with + reactions:

4

Total no. in group:

18

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

intradermal induction: 0.1%; epicutaneous induction: 15%; epicutaneous challenge: 5%

No. with + reactions:

4

Total no. in group:

19

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

intradermal induction: 0.1%; epicutaneous induction: 15%; epicutaneous challenge: 5%

No. with + reactions:

5

Total no. in group:

19

Local skin reactions:

Intradermal injection of Freund's adjuvant (FCA) showed typical skin reactions in all animals. Injection of the test item in the absence of FCA was associated with slight to well defined skin reactions in the animals, however, the skin reactions were less severe than those caused by FCA injection or by injection of test item in the presence of FCA.

After topical induction, bloody skin reactions were noted at the test areas treated with FCA, which turned into necrotic or scab skin reactions thereafter.

after challenge, no skin reactions were noted in test item- or vehicle control-treated animals at any reading time point.

Systemic toxicity:

1/20 control animals died on study Day 14 and 1/20 control animals was not treated due to illness. 1/20 test item treated animals showed strong skin reactions in the neck area and was sacrificed for humane reasons after the topical induction.

Body weight development:

Test item-treated animals showed a reduced body weight development when compared to control animals. The finding was explained by reduced food consumption during the topical induction, which was attributed to impairment caused by the dressing.

Table 1: Body weight and body weight gain (mean ± SD) during the skin sensitisation study in guinea pigs

	BW Day 0	BW Week 1	BW Week 3	BWG
Control	314.6 ± 33.7	365.4 ± 37.5	460.5 ± 49.1	147.8 ± 31.3
Test item	323.1 ± 26.6	375.0 ± 35.7	458.7 ± 47.4	136.6 ± 30.1
BW: Body weight; BWG: Body weight gain

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Data on skin sensitisation are available for alcohols, C12-14, ethoxylated (CAS No. 68439-50-9, EC No. 500-213-3) as well as several member substances of the Alcohol Ethoxylates (AE) category.

 

Studies with alcohols, C12-14, ethoxylated

The skin sensitising potential of alcohols, C12-14, ethoxylated (CAS No. 68439-50-9, EC No. 500-213-3) was assessed in a Guinea Pig Maximisation Test (GPMT) according to OECD guideline 406 observing GLP conditions (Kao, 1996b). Ten test and five control animals (female Dunkin-Hartley guinea pigs) were used for the main study. Based on the results of sighting tests, a concentration of 1% (w/v) of the test substance in arachis oil BP for the intradermal induction was chosen. For the topical induction the test substances was used undiluted and the topical challenge was performed with 25% and 50% (v/v) in arachis oil BP. The test material was considered as a non-sensitiser to guinea pig skin.

The result of the first study is supported by another GMPT performed according to OECD guideline 406 under GLP conditions (BASF, 1988b). First the minimum irritant concentration for the induction applicaton (intracutaneous and epidermal) and the maximum non irritant concentration for the challenge applicaton were estimated in separate pre-tests. For the main test, 20 female guinea pigs (Pirbright white) each  for the test and for the control group were used. The test substance was applied as an aqueous solution for the intracutaneous as well as for the epicutaneous application. 24 and 48 h after removing the patches of the challenge application (2.5% aqueous solution), the test substance caused no skin reaction on the test and on the control animals. According to these findings, the test substance is regarded as a non-sensitiser.

 

Studies in the AE category

Studies investigating skin sensitisation are available for the following AE substances:

 

CAS No.	EC No.	Substance	Study protocol	Hazard conclusion
27252-75-1	500-058-1	Octan-1-ol, ethoxylated	OECD 442E	Negative in the activation of dendritic cells
			OECD 442D	Positive for keratinocyte activation
68439-50-9	500-213-3	Alcohols, C12-14, ethoxylated	OECD 406	Not sensitising
			Similar OECD 406	Not sensitising
68439-49-6	939-518-5	Alcohols, C16-18 (even numbered), ethoxylated, < 2.5 EO	OECD 406	Not sensitising
66455-14-9	500-165-3	Alcohols, C12-13, ethoxylated	Similar OECD 406	Not sensitising

 

Evaluation of skin sensitisation as observed in studies

All available study results indicate that AE substances have no potential to induce skin sensitisation. None of the category member substances turned out to be sensitising to skin in in vivo investigations (GPMT and Buehler test design), thus demonstrating similar toxicological properties. The maximum number of positively induced animals was 5 out of a group of 19 animals in the second study performed with alcohols, C12-14, ethoxylated (CAS No. 68439-50-9, EC No. 500-213-3). No animal showed a positive response in any of the other in vivo studies. The in vitro studies with octan-1-ol, ethoxylated (CAS No. 27252-75-1, EC No. 500-058-1) resulted in an inconclusive picture. A further investigation would be needed to finally conclude on the in vitro skin sensitisation potential of the substance. However, no further testing for the substance is warranted as it is corrosive to skin. In conclusion, the inconclusive assessment for octan-1-ol, ethoxylated (CAS No. 27252-75-1, EC No. 500-058-1) is not considered to invalidate the general findings of the in vivo studies, especially as this substance exhibits corrosive properties. Based on all available data on skin sensitisation in the AE category, it is established that the AE substances are not sensitising to skin.

This evaluation is considered sufficiently conclusive for the hazard assessment and classification and labelling of the AE substances. For a detailed evaluation of the skin sensitisation potential of the substances in the AE category, please refer to the category justification attached to the category object.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The available data on skin sensitisation obtained with alcohols, C12-14, ethoxylated (CAS No. 68439-50-9, EC No. 500-213-3) and with other members of the Alcohol Ethoxylates (AE) category do not meet the criteria for classification according to the CLP Regulation (EC) No. 1272/2008 and are therefore conclusive but not sufficient for classification.