pentasodium 3,7,11-tris(carboxylatomethyl)-15-(C12-18)-alkyl-3,7,11,15-tetraazaheptadecane-1,17-dioate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 May 1995 - 14 June 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was performed similar to OECD guidelines and under GLP. No data on substance identity (SURFACTANT HDC 94-05-20-03). Concentrations did not elicit required irritation levels

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Principles of method if other than guideline:

The induction procedure was repeated for three weeks to give a total of nine 6-hour exposure instead of three 6 hour induction exposures.

GLP compliance:

yes (incl. QA statement)

Type of study:

Buehler test

Justification for non-LLNA method:

The default animal study for skin sensitisation in the ECHA Guidelines, is the Local Lymph Node Assay (LLNA) OECD429. However it is accepted in the OECD guideline for the LLNA that it has some limitations, as follows: “Despite the advantages of the LLNA over TG 406, it should be recognised that there are certain limitations that may necessitate the use of TG 406 (13) (e.g. false negative findings in the LLNA with certain metals, false positive findings with certain skin irritants [such as some surfactant type chemicals] (19) (20), or solubility of the test substance)”. For the amphoteric glycinate substances, there are available in-vivo skin sensitizing studies performed before the LLNA was recommended as a standard. As these studies are considered to be of high reliability rating and the results are considered relevant, it is not justified from an animal well-fare perspective to perform any additional skin sensitising studies. For this reason the available guinea pig studies are provided as key studies.

Test material

Test material information

Constituent 1

Reference substance name:

Amines, N-C8-22-alkylpolytrimethylenepoly-, carboxymethyl derivs., sodium salts

EC Number:

307-458-3

EC Name:

Amines, N-C8-22-alkylpolytrimethylenepoly-, carboxymethyl derivs., sodium salts

Cas Number:

97659-53-5

Test material form:

other: Aqueous solution

Details on test material:

Sponsor's identification : SURFACTANT HDC 94-05-20-03
Date received : 27 March 1995
Description : straw coloured liquid
Storage conditions : room temperature in the dark

(e-mail attached explaining that SURFACTANT HDC 94-05-20-03 = Ampholak 7CX:
CAS 97659-53-5
Cocoamphopolycarboxyglycinate)

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: by David Hall
Limited, Burton-on-Trent, Staffordshire, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 300-387 grams
- Housing: single or in pairs in solid-floor polypropylene cages furnished with woodflakes.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: minumum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 51-64
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 10 May 1995 - 14 June 1995

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

epicutaneous, occlusive

Vehicle:

other: unchnged or distilled water

Induction 2

Concentration / amount:

Topical Induction
Group 1 : undiluted as supplied
Group 2 : 25% v/v in distilled water
Topical Challenge : undiluted as supplied and 25% v/v in distilled water

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: unchnged or distilled water

Challenge 2

Concentration / amount:

Topical Induction
Group 1 : undiluted as supplied
Group 2 : 25% v/v in distilled water
Topical Challenge : undiluted as supplied and 25% v/v in distilled water

No. of animals per dose:

20 test animals per group and 10 control animals

Details on study design:

RANGE FINDING TESTS:

Selection of Concentration for Topical Induction
Two previously untreated guinea pigs were treated with 0.5 ml of the undiluted test material and three concentrations of the test material in 80% aqueous ethanol (75%, 50%, and 25% v/v). Applications were made to the clipped flanks under occlusive dressings for an exposure period of 6 hours. The degree of erythema and oedema was evaluated 24 and 48 hours after dressing removal. The highest concentration of the test material producing
only mild dermal irritation was selected for the topical induction of Group 1 animals in the main study. A second concentration (25% v/v in distilled water) was selected by the study sponsor for induction of Group 2 animals.

Selection of Concentration for Topical Challenge
Two guinea pigs were treated with 0.5 ml of undiluted test material and one concentration of the test material in distilled water (75% v/v). These animals had been treated identically to the control animals of the main study on Days 0, 7 and 14. Applications were made to the clipped flanks under occlusive dressings for an exposure period of 6 hours. The degree of erythema and oedema was evaluated 24 and 48 hours after dressing removal. The highest concentration of the test material which produced no evidence of dermal irritation, was selected for the topical challenge stage of the main study. 25% v/v was selected by the study sponsor as the lower concentration for the topical challenge stage of the main study.

MAIN STUDY
A. INDUCTION EXPOSURE
The hair was removed from an area on the left flank of each animal with veterinary clippers. A quantity of 0.5 ml of the undiluted test material was applied to the shorn left flank on an absorbent cotton lint patch (approximate size 15 mm x 35 mm). The patch was held in place under a strip of
surgical adhesive tape (BLENDERM: approximate size 50 mm x 60 mm) and covered with an overlapping length of aluminium foil. The patch and foil were further secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 70 mm x 250 mm) wound in a double layer around the torso of each animal.
- No. of exposures: 9
- Exposure period: 6 hours
- Test groups: 2x 20 animals
- Control group: 10 animals
- Site: left flank
- Frequency of applications: lnduction was performed on days 0; 2, 4, 7, 9, 1 1, 14, 16 and 18.
- Duration: 3 weeks
- Concentrations: 25% v/v in distilled water or unchanged as suppplied

lnduction of the Control Animals: The topical applications followed the same procedure as for the test animals except that the vehicle alone was applied.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 28
- Exposure period: 6 hours
- Test groups: Shortly before treatment on Day 28, an area approximately 50 mm x 70 mm on the right shorn flank of each animal, was clipped free of hair with veterinary clippers. A quantity of 0.5 ml of the undiluted test material was applied to the shorn right flank of each animal on an absorbent cotton lint patch (approximate size 15 mm x 30 mm). The patch was held in place by a strip of surgical adhesive tape (BLENDERM: approximate size 40 mm x 50 mm). The test material at a concentration of 25% v/v in distilled water was similarly applied to a separate skin site on the right shorn flank. The patches were covered with an overlapping length of aluminium foil and further secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 70. mm x 250 mm) wound in a double layer around the torso.
- Control group: the same procedure as for the test animals
- Site: left flank
- Concentrations: 25% v/v in distilled water or unchanged as suppplied
- Evaluation (hr after challenge): Approximately 24 and 48 hours after dressing removal, the degree of erythema and oedema was quantified

OTHER: -

Challenge controls:

The same procedure as for the test animals, see above.

Positive control substance(s):

not required

Results and discussion

In vivo (non-LLNA)

Results

Reading:

other: all readings

Group:

other: all test groups

Dose level:

25% or undiluted as supplied

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: other: all readings. Group: other: all test groups. Dose level: 25% or undiluted as supplied. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Any other information on results incl. tables

Skin Reactions Observed After Topical lnduction

One animal was killed for humane reasons on day 13 due to respiratory problems, and one animal was found dead on day 16 (the cause of death was not investigated). The absence of these animals did not affect the purpose or integrity of the study.

Group 1 (Undiluted as Supplied)

Very slight to well defined erythema and very slight to slight oedema were elicited by the test material. Other skin reactions noted were desquamation, hardened light brown-coloured scab, hardened dark browdblack-coloured scab and small superficial scattered scabs. On occasions the skin reactions prevented the accurate evaluation of oedema and/or erythema. On occasions the test sites were changed due to severe reactions.

Group 2 (25% v/v in Distilled Water)

One test group animal was killed for humane reasons on day 13. One test group animal was found dead on day 16. The cause of death was not determined. The absence of these animals was considered not to affect the purpose or integrity of the study. Very slight erythema and very slight oedema were elicited by the test material. Other skin reactions noted were desquamation, hardened light brown-coloured scab and small superficial scattered scabs. On one occasion desquarnation prevented accurate evaluation of erythema in one test group animal. On occasions the test sites were changed due to severe reactions.

Vehicle Control

No skin reactions were noted at the vehicle control sites of control group animals following topical induction.

Skin Reactions Observed After Topical Challenge

No skin reactions were noted at the challenge sites of the test or control group animals at the 24 or 48-hour observations.

Bodyweight Individual

Bodyweight gains of guinea pigs in the test group, between Day 0 and Day 30, were comparable to those observed in the control group animals over the same - period.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

The test material, consisting of 40% active ingredient and 60% water, produced a 0% (0/20) sensitisation rate and was classified as a non-sensitiser to guinea pig skin under the conditions of the study. Therefore the active ingredient Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) is considered to be non sensitising.

Executive summary:

A study was performed to assess the contact sensitisation potential of the test material in the albino guinea pig. The study was performed in compliance with the OECD Guidelines for Testing of Chemicals No. 406 "Skin Sensitisation" B6 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC). Two groups of twenty test animals and ten control animals were used for the main study. Based on the results of sighting tests, the concentrations of test material for the induction and challenge phases were selected as follows: Topical Induction Group 1 : undiluted as supplied Group 2 : 25% v/v in distilled water Topical Challenge : undiluted as supplied and 25% v/v in distilled water. No effects were observed. The test material, consisting of 40% active ingredient and 60% water, produced a 0% (0/20) sensitisation rate and was classified as a non-sensitiser to guinea pig skin under the conditions of the study. Therefore the active ingredient Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) is considered to be non sensitising.