Alcohols, C11-15-secondary, ethoxylated

Endpoint summary

Description of key information

Experimental data obtained from a local lymph node assay and a guinea-pig maximisation test for skin sensitisation potential of Softanol 30 were discordant. A weight of evidence assessment of skin sensitisation potential taking into account available evidence concluded that Softanol 30 is unlikely to be a skin sensitiser. Therefore classification of Softanol 30 as a skin sensitiser is not proposed.

There are no data on the respiratory sensitisation potential of Softanol 30. No tests are available which will identify respiratory sensitisation hazard. No human data have been found which suggest a respiratory sensitisation hazard. No classification for respiratory sensitisation is proposed.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation, other

Remarks:

existing in vivo study

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

12 May to 10 June 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in accordance with an internationally accepted guideline (OECD)

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
-Source: United Kingdom
- Age at study initiation: approximately 8- 12 weeks
- Weight at study initiation: 17.3 to 22.2 g
- Housing: individually in polycarbonate cages with woodflake bedding.
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): freely available
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23°C
- Humidity (%): 40 to 70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: 18 May to 9 June 2010

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

5, 10 and 25% v/v

No. of animals per dose:

4 females/dose

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: A vehicle trial was conducted and Softanol 30 showed that it formed a clear solution at 50% v/v in acetone:olive oil (4:1 v/v), which was satisfactory for dose administration.
- Irritation: signs of irritation were seen over the dosed area on Days 2 to 4 (100% v/v) and Days 2 and 3 (Group 50%).
- Lymph node proliferation response: no data

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Pooled treatment group approach
- Criteria used to consider a positive response: the test substance is regarded as a sensitizer if at least one concentration of the test substance
results in a three-fold greater increase in 3HTdR incorporation compared to control values

TREATMENT PREPARATION AND ADMINISTRATION:
- Preliminary test: Three females were treated with one of three concentrations of the test substance. The mice were treated by daily application of 25 μl of appropriate concentration of the test substance to the dorsal surface of each ear for three consecutive days (Days 1-3).
- Main test: Groups of four mice were treated at one of three concentrations of the test substance. The mice were treated by daily application of 25 μl of the appropriate concentration of the test substance to the dorsal surface of each ear for three consecutive days (Days 1-3).
In the main phase of the study, five days following the first topical application of test substance (Day 6) all mice were treated with 250 μl of phosphate buffered saline containing 3H-methyl Thymidinea (3HTdR: 80 μCi/mL) giving a nominal 20 μCi to each mouse.

A single cell suspension of lymph node cells (LNC) was prepared by gentle mechanical disaggregation through a stainless steel gauze (200 mesh size). The pooled LNC were then washed by adding 10 mL phosphate buffered saline, pelleted at 190 x g for 10 minutes and resuspended. The cells were washed twice again and resuspended in 3 mL trichloroacetic acid (TCA: 5%) following the final wash. After overnight incubation with 5% TCA at 4°C, the precipitate was recovered by centrifugation and resuspended in 1 mL 5% TCA and transferred to 10 mL Ultima gold scintillation fluid on Day 7. 3HTdR incorporation was measured by β-scintillation counting.

Positive control results:

Responses to the positive control substance hexyl cinnamic aldehyde (HCA), in contemporaneous studies demonstrated the reliability and sensitivity of this assay to detect skin sensitization potential in this laboratory.

Parameter:

SI

Remarks on result:

other: 5% v/v 1.6 10% v/v 1.7 25% v/v 5.1

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: 5% v/v 3994.75 10% v/v 4477.95 25% v/v 13141.35

The test substance is regarded as a sensitizer if at least one concentration of the test substance results in a three-fold greater increase in 3HTdR incorporation compared to control values.

Interpretation of results:

sensitising

Remarks:

Migrated information

Conclusions:

Softanol 30 is regarded as a potential skin sensitizer.

Executive summary:

In a dermal sensitization study realised according to the OECD guideline 429 and in compliance with GLP (HLS 2010, PLZ0020), Softanol 30 in acetone/olive oil (4:1 v/v) at 5, 10 and 25 % was tested on female CBA mice (4/group), using the LLNA (Local Lymph Node Assay) . 

Animals were treated by daily application of 25µl of the appropriate concentration to the dorsal surface of each ear for 3 consecutive days and all mice were treated with 250 μl of phosphate buffered saline containing 3H-methyl Thymidine (3HTdR: 80 μCi/mL) giving a nominal 20 μCi to each mouse on Day 6.

Stimulation index were 1.6, 1.7 and 5.1 at 5, 10 and 25%. Therefore Softanol 30 is regarded as a potential skin sensitizer.

Reference 2

Endpoint:

skin sensitisation, other

Remarks:

exisiting in vivo study

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

This study has been disregarded (see below for details) and a second test for the evaluation of the sensitization properties of Softanol 30 has been realised (LLNA). 1. Intradermal induction concentrations too low: The study has been realised according to the OECD406 guideline from 1992 and to the Magnusson of Kligman literature. In this article, the maximal concentration sufficient for the intradermal injection was 5%. According to the recent version of the OECD406 guideline, doses selected should be the highest concentration causing mild-to-moderate skin irritation and the highest non-irritant concentration for the challenge. From a theoretical point of view, liquid can be tested at concentrations up to 75% or more. 2. Epicutaneous induction concentrations too low: In the study, concentration of 75 % has been used. But if the results between 75% and 100% were comparable, the highest concentration should have been used for the epidermal induction. Based on these two observations, the concentrations seem to be not sufficiently high to ensure that the animals were correctly sensitized. And the results can therefore be equivocal and possibly false negative.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

test of doses very low

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Species:

guinea pig

Strain:

other: lbm: GOHI; SPF·quality guinea pigs (synonym: Himalayan spotted)

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Woelferstrasse 4, 4414 Fuellinsdorf / Switzerland
- Age at study initiation: 5 - 7 weeks
- Weight at study initiation: Control and Test Group: 339 - 460g; Pretest: 399 - 436 g
- Housing: Individually in Makrolon type-3 cages with standard Softwood bedding.
- Diet (e.g. ad libitum): ad libitum Pelleted standard Nafag Ecosan 845 25W4, batch nos. 28/96 and 32/96 gu1nea pig breeding/maintenance diet ('Nafag', Naehr-und Futtermittel AG, CH-9202 Gossau).
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid (1 g/l) via the drinking water.
- Acclimation period: 1week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

other: ethanol

Concentration / amount:

Intradermal induction: 5%
Epicutaneous induction: 75%
Epicutaneous challenge: 25%

Route:

epicutaneous, occlusive

Vehicle:

other: ethanol

Concentration / amount:

Intradermal induction: 5%
Epicutaneous induction: 75%
Epicutaneous challenge: 25%

No. of animals per dose:

Control group: 10 females
Tests groups: 20 females

Details on study design:

RANGE FINDING TESTS:
Intradermal and epidermal pretests were realised on 2 and 4 animals, respectively.

- INTRADERMAL INJECTIONS: Intradermal injections (0.1 ml/site) were made into the clipped flank of two guinea-pigs at concentrations of 5, 3 and 1% of the test article in ethanol. The resulting dermal reactions were assessed 24 hours later. For intradermal induction application a 5% test article dilution was selected.
- EPIDERMAL APPLICATIONS: Both flanks of each of 4 guinea pigs were clipped and shaved just prior to the application. Thereafter 4 patches of filter paper (2 x 2 cm) were saturated with the test article at 100% (A), 75% (B), 50% (C) and 25% (D) and applied to the clipped and shaved flanks. Ethanol was used for the dilutions. The patches were covered by a strip of aluminium foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. This procedure ensured the intensive contact of the test article. The dressings were removed after an exposure period of 24 hours. The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema on a numerical basis according to Draize.


MAIN STUDY
A. INDUCTION EXPOSURE
INTRADERMAL INDUCTION
- No. of exposures: 1
- Exposure period: day 0
- Test groups:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) The test article, diluted to 5% with ethanol.
3) The test article diluted to 5% by emulsion in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.

- Control group:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) Ethanol.
3) 1:1 (v/v) mixture of ethanol in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.

- Site: clipped dorsal skin from the scapular region (approximately 6 x 8 cm)
- Frequency of applications: once
- Duration: one injection only
- Concentrations: 5%

EPICUTANEOUS INDUCTION
- No. of exposures: 1
- Exposure period: day 7
- Test groups: mixture of Softanol 30 in ethanol
- Control group: ethanol only
- Site: clipped dorsal skin from the scapular region (approximately 6 x 8 cm)
- Frequency of applications: 1
- Duration: 48 h
- Concentrations: 75% in ethanol


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 21
- Exposure period: 24 hours
- Test groups: left flank at 25% of Softanol 30 in ethanol
- Control group: right flank with ethanol only
- Site: clipped skin of the flank (approximately 5 x 5 cm)
- Concentrations: 25%
- Evaluation (hr after challenge): 24 and 48 hours after removal of dressing.

Challenge controls:

None

Positive control substance(s):

yes

Remarks:

The positive controls were performed with 2·MERCAPTOBENZOTHIAZOLE (RCC project 900731) and ALPHA-HEXYLCINNAMALDEHYDE (RCC project 900742) and performed from 16-0CT-1995 to 16-NOV-1995 and from 17-OCT-1995 to 17-NOV-1995 respectively.

Positive control results:

2-mercaptobenzothiazole gave 18/19 positive animals (95%) positive response.
alpha-cinnamaldehyde gave 15/20 positive animals (75%) positive response.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

25% in ethanol

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% in ethanol. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25% in ethanol

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% in ethanol. No with. + reactions: 0.0. Total no. in groups: 20.0.

No symptoms of systemic toxicity were observed in the animals.

The body weight of the animals was within the physiological range of variability known for this strain and age. However, one animal of the epidermal pretest lost weight during its treatment period.

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

SOFTANOL 30 applied at a concentration of 25% in ethanol is considered to be a non sensitizer when used under the described test conditions.

Executive summary:

The purpose of this skin sensitizing study was to assess the possible allergenic potential of SOFTANOL 30 to albino guinea pigs. The maximalisation test of Magnusson and Kligman (1969) was used. Twenty female animals of the test group were induced intradermally once in the first week and epidermally once in the second week with SOFTANOL 30 at 5% and 75% in ethanol, respectively.

Two weeks after the epidermal induction application, the animals were challenged with the vehicle ethanol and the same test substance used for induction at the highest non-irritating concentration of 25% in ethanol.

The animals of the control group were induced with ethanol only and treated once at challenge with ethanol and SOFTANOL 30 at 25% in ethanol.

 

In this study 0% of the animals of the test and control group were observed with positive skin reactions after treatment with a non-irritant test substance concentration of 25% in ethanol.

A response of at least 30%; positive animals is considered positive "R43": may cause sensitization by skin contact according to the 'Commission Directive 93/21/EEC, April 27, 1993 adapting to technical progress for the 18th time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous Substances".

Therefore, the test article SOFTANOL 30 applied at a concentration of 25% in ethanol is considered to be a non sensitizer when used under the described test conditions.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

Three experimental studies are available: a Local Lymph Node Assay conducted in 2010, a Guinea-Pig Maximalisation Test conducted in 1996, and a human patch study performed in 1972. The human study has been disregarded for the reasons given below.

In a dermal sensitization study conducted according to the OECD guideline 429 and in compliance with GLP (HLS 2010, PLZ0020), Softanol 30 in acetone/olive oil (4:1 v/v) at 5, 10 and 25 % was tested on female CBA mice (4/group), using the LLNA (Local Lymph Node Assay) . 

Animals were treated by daily application of 25µl of the appropriate concentration to the dorsal surface of each ear for 3 consecutive days and all mice were treated with 250 μl of phosphate buffered saline containing 3H-methyl Thymidine (3HTdR: 80 μCi/mL) giving a nominal 20 μCi to each mouse on Day 6.

Stimulation index were 1.6, 1.7 and 5.1 at 5, 10 and 25%. Therefore Softanol 30 is regarded as a potential skin sensitizer.

  

A Magnusson and Kligman test is also available (RCC 1996, report 622427),

The maximalisation test of Magnusson and Kligman (1969) was conducted on Softanol 30 on albino guinea pigs. Twenty female animals of the test group were induced intradermally once in the first week and epidermally once in the second week with SOFTANOL 30 at 5% and 75% in ethanol, respectively.

Two weeks after the epidermal induction application, the animals were challenged with the vehicle ethanol and the same test substance used for induction at the highest non-irritating concentration of 25% in ethanol.

In this study 0% of the animals of the test and control group were observed with positive skin reactions after treatment with a non-irritant test substance concentration of 25% in ethanol. Therefore, Softanol 30 applied at a concentration of 25% in ethanol is considered to be a non sensitizer when used under the described test conditions.

 

The third study, a patch test using five human volunteers with a test concentration of 0.6 wt % is disregarded because of a lack of information on methodology, interpretation of results and the scoring scale used.

Interrogation of the molecule by Toxtree v2.6.13 and OECD toolbox v 3.2 triggered no alerts for skin sensitisation.

Read-across using skin sensitisation data for non-ionic surfactants in OECD toolbox v 3.2 predicted CAS 68131 -40 -8 to be a non-sensitiser.

Cross-reference section 13 Assessment Reports for sensitisation expert report.

It should be noted, also, in any production sites in Japan, since 1972, no workers have suffered from any adverse effects including a skin rash.

Justification for classification or non-classification

When applied a 25% on mice ears, Softanol 30 induced a stimulation of the lymphocyte proliferation: 5.1 when compared to control. In a GPMT Softanol 30 did not produce a positive response. A weight of evidence assessment including (Q)SAR and read-across concluded that Softanol 30 is not a skin sensitiser. The LLNA result is a false positive related to its surfactant properties. Softanol 30 is therefore not regarded as a potential skin sensitizer and should not be classified as such.

Cross-reference section 13 Assessment Reports for sensitisation expert report.