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

Endpoint summary

Description of key information

One modified Bühler test on Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and one Guinnea pig maximization test on Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9) are available within the amphoteric, glycinate substance group. The results indicate lack of sensitisation, although the testing had not been performed with pure compounds, but the technical 40% aqueous solutions. However, if the pure freeze-dried substances were to be tested, testing of solids in LLNA generally results to about 50% concentrations, which is not far from the available data using 40%. Based on animal welfare grounds further testing at higher concentrations was therefore not proposed.

 

Profiling the amphoteric, glycinate substances for skin sensitizing properties using Derek Nexus, TOPKAT as well as the QSAR Toolbox indicates that no alerts are found for protein binding or structural alerts. The query structure does not contain any unclassified or misclassified features and is consequently predicted to be a non-sensitiser. There are no reports on incidences of sensitisation from industrial production and use of the substances. Taken all the available information together, read across using the available data is considered applicable within the substance group.

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:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

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.

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

Route:

epicutaneous, occlusive

Vehicle:

other: unchnged or distilled water

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

Route:

epicutaneous, occlusive

Vehicle:

other: unchnged or distilled water

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

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.

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.

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.

Reference 2

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

One modified Bühler test on Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and one Guinnea pig maximization test on Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9) are available within the amphoteric, glycinate substance group. The results indicate lack of sensitisation, although the testing had not been performed with pure compounds, but the technical 40% aqueous solutions. However, if the pure freeze-dried substances were to be tested, testing of solids in LLNA generally results to about 50% concentrations, which is not far from the available data using 40%. Based on animal welfare grounds further testing at higher concentrations was therefore not proposed.
 
Profiling the amphoteric, glycinate substances for skin sensitizing properties using Derek Nexus, TOPKAT as well as the QSAR Toolbox indicates that no alerts are found for protein binding or structural alerts. The query structure does not contain any unclassified or misclassified features and is consequently predicted to be a non-sensitiser. There are no reports on incidences of sensitisation from industrial production and use of the substances. Taken all the available information together, read across using the available data is considered applicable within the substance group.

Key result

Reading:

other: Challenge phase

Hours after challenge:

24

Group:

test chemical

Dose level:

100% test material (corresponds to 40% a.i)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Not described

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

other: Challenge phase

Hours after challenge:

48

Group:

test chemical

Dose level:

100% test material (corresponds to 40% a.i.)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Not described

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

other: Challenge phase

Hours after challenge:

24

Group:

test chemical

Dose level:

50% test material (corresponds to 20% a.i.)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Not described

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

other: Challenge phase

Hours after challenge:

48

Group:

test chemical

Dose level:

50% test material (corresponds to 20% a.i.)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Not described

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

other: Challenge phase

Hours after challenge:

24

Group:

negative control

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Not described

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

other: Challenge phase

Hours after challenge:

48

Group:

negative control

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Not described

Remarks on result:

no indication of skin sensitisation

No reaction was observed in any of the animals exposed to the test substance or in the control group.

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

On challenge with the test material, consisting of 40% active ingredient and 60% water, no visible response was exhibited by any animal in the test or control group when challenged with the undiluted test material and 50% aqueous concentration of the test material. From the results of this study there was no evidence to suggest that the active ingredient Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9) acts as a sensitiser in the guinea pig.

Executive summary:

Accoring to methods similar to OECD 406 and under GLP ten guinea pigs were treated by intradermal injection in the shoulder region with the test material, Freund's Complete Adjuvant and a mixture of the test material and Freund's Complete Adjuvant. Seven days later this induction procedure was boosted by the topical application of the test material over the injection site. A second group of ten animals were similarly treated but distilled water was substituted for the test material. Two weeks after the induction phase all animals of both test and control groups were challenged with two concentrations of the test material applied topically to the flanks. On challenge with the test material no visible response was exhibited by any animal in the test or control group when challenged with the undiluted test material and 50% aqueous concentration of the test material. From the results of this study there was no evidence to suggest that the active ingredient of the test material, Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9) acts as a sensitiser in the guinea pig.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

There are no guidelines for an animal test for respiratory sensitization, however in general respiratory sensitizers are also skin sensitizers. Based on the available study on Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5), as well the read across data within the group of amphoteric, glycinate substance group the substance was not found to be a skin sensitizer. This indicates that the substance is unlikely to possess any significant potential for respiratory sensitization. Furthermore Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) is an aqueous solution with a low vapour, therefore inhalation exposure is unlikely.

Justification for classification or non-classification

Skin

Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and the other three substances within the amphoteric, glycinate substane group, are not found to be skin sensitizers. This is based on the two available in-vivo studies, one performed on Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and the other on Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9). Together with the available QSAR data, the overall conclusion is that the four amphoteric glycinate substances have low potential for skin sensitising properties.

Inhalation 

Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) has no skin sensitising properties. This together withthe overall information on the amphoteric glycinate substancesindicates that the substance is unlikely to possess any significant potential for respiratory sensitization. All of the amphoteric glycinate substances are produced and handled as aqueous solutions with low vapour pressures, therefore inhalation exposure is unlikely. Data on acute inhalation is lacking, but taken the result from the skin sensitization study and the low potential for inhalation exposure into consideration, the substance it is not classified as a respiratory sensitizer.