Butanedioic acid, sulfo-, 4-[2-[(2-hydroxyethyl)amino]ethyl] ester, N-C18-unsatd. acyl derivs., disodium salts

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

2012-2013

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study has been conducted according to GLP and valid methods, therefore it is considered to be adequate, reliable and relevant for classifcation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Stolzenseeweg 32-36, 88353 Kißlegg, Germany
- Age at start of administration: 25 days
- Weight at start of administration: 251 – 295 g (excluding positive control group); 316 – 370 g (positive control group)
- Housing: In pairs in MAKROLON cages (MZK 80/25); Granulated textured wood (Granulat A2, J. BRANDENBURG, 49424 Goldenstedt, Germany) was used as bedding material in the cages. The cages were changed and cleaned twice a week.
- Diet (e.g. ad libitum): Commercial diet, ssniff® Ms-H (ssniff Spezialdiäten GmbH, 59494 Soest, Germany , ad libitum
- Water (e.g. ad libitum): Tap water in drinking bottles was offered ad libitum.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C (maximum range)
- Humidity (%): 55% ± 15% (maximum range)
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: November 16, 2012 To: February 23,2013

Route:

other: lntracutaneous(stage 1) and Topical (stage 2)

Vehicle:

other: aqua ad iniectabilia

Concentration / amount:

Preliminary test:
Stage1: intracutaneous: 10, 5, 1, 0.5, 0.1 and 0.01% suspension of the test item in aqua ad iniectabilia
Stage 2: topical: 100, 75, 50, 25, 10, 5 and 1% suspension of the test item in aqua ad iniectabilia
Stage 3: topical: 100, 75, 50, 25, 10, 5 and 1% suspension of the test item in aqua ad iniectabilia

Main test:
Stage1: intracutaneous: 0.5% suspension of the test item in aqua ad iniectabilia
Stage 2: topical: 25% suspension of the test item in aqua ad iniectabilia
Stage 3: topical: 10% suspension of the test item in aqua ad iniectabilia

Route:

other: Topical (stage 3)

Vehicle:

other: aqua ad iniectabilia

Concentration / amount:

Preliminary test:
Stage1: intracutaneous: 10, 5, 1, 0.5, 0.1 and 0.01% suspension of the test item in aqua ad iniectabilia
Stage 2: topical: 100, 75, 50, 25, 10, 5 and 1% suspension of the test item in aqua ad iniectabilia
Stage 3: topical: 100, 75, 50, 25, 10, 5 and 1% suspension of the test item in aqua ad iniectabilia

Main test:
Stage1: intracutaneous: 0.5% suspension of the test item in aqua ad iniectabilia
Stage 2: topical: 25% suspension of the test item in aqua ad iniectabilia
Stage 3: topical: 10% suspension of the test item in aqua ad iniectabilia

No. of animals per dose:

Preliminary test: 10 animals in total
8 animals for the topical administration
2 animals for the intracutaneous administration

Main test: 15 animals in total (20 animals historical data)
10 test group
5 vehicle control group
20 positive control group (historical)

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE

Stage 1: Intradermal injections Day 0
- No. of exposures: 3 (pairs of injections)
- Exposure period: Single injections
- Test groups: 10 animals, 0.5% concentration test item in aqua ad iniectabilia
Three pairs of intradermal injections of 0.1 mL were given in the shoulder region which was cleared of hair so that one of each pair lay on each side of midline.
(1) Freund's complete adjuvant
(diluted 1 : 1 with 0.9% NaCl )
(2) the test item (concentration see above)
(3) the test item in a 1:1 mixture (v/v) FCA/physiological saline
In injection 3, the final concentration of the test item was equal to that used in injection 2.
Injections (1) and (2) were given close to each other and nearest the head, while (3) was given towards the caudal part of the test area.
- Control group:
Vehicle control: 5 animals:
(1)Freund's complete adjuvant (diluted 1 : 1 with 0.9% NaCl )
(2) aqua ad iniectabilia
(3) aqua ad iniectabilia in a 1:1 mixture (v/v) FCA/physiological saline
Positive control: 20 animals (historical data): 10% (v/v) α-hexyl cinnamic aldehyde solution
- Site: Shoulder region
- Frequency of applications: Single injections
- Duration: Single injections (Day 0) and six days interval before stage 2 (Day 7)
- Concentrations: 0.5% concentration test item in tap water

Stage 2: Topical Day 7
- No. of exposures: 1 (patch)
- Exposure period: 48 hours
- Test groups: 10 animals: 25% concentration test item in aqua ad iniectabilia
- Control group:
Vehicle control: 5 animals: aqua ad iniectabilia
Positive control: 20 animals (historical data): undiluted α-hexyl cinnamic aldehyde
- Site: Shoulder region
- Frequency of applications: 1
- Duration: 48 hours
- Concentrations: 25% concentration test item in aqua ad iniectabilia


B. CHALLENGE EXPOSURE
Stage 3: Topical: Day 21
- No. of exposures: 1 (patch)
- Day(s) of challenge: Day 21
- Exposure period: 24 hours
- Test groups: The filter paper containing the test item (10% suspension of the test item in aqua ad iniectabilia) was applied to the left flank, the filter paper with the vehicle to the right flank of the animal.
- Control group:
Vehicle control: The left flank was treated with the test item (10% concentration in aqua ad iniectabilia), the right flank with the vehicle i.e. in the same way as the test group.
Positive control: The filter paper containing 0.01% α-hexyl cinnamic aldehyde solution was applied to the left flank, the filter paper with the vehicle to the right flank of the animal.
- Site: Flanks
- Concentrations: 10% suspension of the test item in tap water
- Evaluation (hr after challenge):
21 hours after removing the filter paper the challenge area was cleaned and cleared of hair if necessary; three hours later (at 48 hours from the start of challenge application) the skin reaction was observed and recorded; 24 hours after this observation a second observation (72 hours) was made and recorded.

OTHER:
MAGNUSSON/KLIGMAN grading scale tor the evaluation of challenge patch test reactions
0 = no visible change
1 = discrete or patchy erythema
2 = moderate and confluent erythema
3 = intense erythema and swelling

Challenge controls:

- Vehicle control: The left flank was treated with the test item (10% concentration in aqua ad iniectabilia), the right flank with the vehicle i.e. in the same way as the test group.
- Positive control: The filter paper containing 0.01% α-hexyl cinnamic aldehyde solution was applied to the left flank, the filter paper with the vehicle to the right flank of the animal.

Positive control substance(s):

yes

Remarks:

α-hexyl cinnamic aldehyde

Positive control results:

Animals of the same strain treated with α-hexyl cinnamic aldehyde in sesame oil exhibited a sensitising reaction in all animals in form of a moderate and confluent erythema (grade 2).

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10% suspension of the test item in aqua ad iniectabilia (2mL)

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10% suspension of the test item in aqua ad iniectabilia (2mL). No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

10% suspension of the test item in aqua ad iniectabilia (2mL)

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 10% suspension of the test item in aqua ad iniectabilia (2mL). No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

aqua ad iniectabilia (2mL)

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: aqua ad iniectabilia (2mL). No with. + reactions: 0.0. Total no. in groups: 5.0.

Reading:

2nd reading

Hours after challenge:

72

Group:

negative control

Dose level:

aqua ad iniectabilia (2mL)

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: aqua ad iniectabilia (2mL). No with. + reactions: 0.0. Total no. in groups: 5.0.

Reading:

1st reading

Hours after challenge:

48

Group:

positive control

Dose level:

0.01% α-hexyl cinnamic aldehyde

No. with + reactions:

20

Total no. in group:

20

Clinical observations:

moderate and confluent erythema left flank

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.01% α-hexyl cinnamic aldehyde . No with. + reactions: 20.0. Total no. in groups: 20.0. Clinical observations: moderate and confluent erythema left flank.

Reading:

2nd reading

Hours after challenge:

72

Group:

positive control

Dose level:

0.01% α-hexyl cinnamic aldehyde

No. with + reactions:

20

Total no. in group:

20

Clinical observations:

moderate and confluent erythema left flank

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: positive control. Dose level: 0.01% α-hexyl cinnamic aldehyde . No with. + reactions: 20.0. Total no. in groups: 20.0. Clinical observations: moderate and confluent erythema left flank.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the present test conditions, test item containing 39.80% active ingredient, was found to be not sensitising to guinea pigs in a test model according to MAGNUSSON and KLIGMAN.

Executive summary:

The purpose of this study was to determine the potential of the test item, containing 39.80% active ingredient, in guinea pigs in a test model according to MAGNUSSON and KLIGMAN.

From a preliminary experiment, it was decided to use a 0.5% concentration in aqua ad iniectabilia for the 1st (intracutaneous) stage, a 25% suspension in aqua ad iniectabilia for the 2nd (topical) induction stage and a 10% suspension in aqua ad iniectabilia for the challenge.

A 0.5% suspension in aqua ad iniectabilia chosen for the 1st (intracutaneous) induction stage revealed a discrete or patchy erythema in all 10 animals 24 and 48 hours after administration. 2 mL of a 25% suspension of the test item in aqua ad iniectabilia/animal chosen for the 2nd (topical) induction stage revealed a moderate and confluent erythema 48 hours and a discrete or patchy erythema 72 hours after start of exposure in all 10 animals. The challenge with 2 mL of a 10% suspension of the test item in aqua ad iniectabilia /animal - the maximum non-irritating concentration - revealed no skin irritation in any animal and, thus, the test item had no sensitising properties.

The vehicle control revealed no skin reactions.

Animals of the same strain treated with α-hexyl cinnamic aldehyde in sesame oil exhibited a sensitising reaction in all animals in form of a moderate and confluent erythema (grade 2).

Behaviour of the animals remained unchanged. Body weights were not affected

Under the present test conditions, test item containing 39.8% active ingredient, was found to be not sensitising to guinea pigs in a test model according to MAGNUSSON and KLIGMAN.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

No test data were available for current substance, however read across data were available from N2 and N3 subgroup members:

-'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts'
- ‘Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite’
-'Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium salts'.

Justification for read across within the category of N-containing sulphosuccinates (N2 and N3 subgroups) is documented in a separate document attached in Section 13.

 

Skin sensitisation was assessed based on various Weight-of-Evidence studies from read-across substances:
- In a skin sensitisation study with read-across substance 'Butanedioic acid, sulfo-, 4-[2-[(2-hydroxyethyl)amino]ethyl] ester, N-C18-unsatd. acyl derivs., disodium salts' according to the method of Magnusson and Kligman, 2 groups (test and control group) of 20 animals each were tested with the registered substance containing 40% active ingredient (Sterner and Chibanguza, 1988). Intradermal induction was done in 10 male and 10 female control and test animals with 0.05 mL. Test group:1 pair (2) injections of test substance 10% solution (4% active ingredient) in deionised water; 1 pair (2) injections of test substance 10% solution in FCA and 1 pair (2) injections of FCA undiluted.Control group:1 pair (2) injections of FCA undiluted; 1 pair (2) injections of dionised water 10% solution in FCA and 1 pair (2) injections of deionised water undiluted.Epicutaneous induction was done, 7 days after intradermal induction; the first dermal treatment started with 0.5 mL of the test substance (50% solution= 20% active ingredient) on the left flank and 0.5 mL vehicle (deionised water) on the right flank in a closed patch test. After 48 hours the dressing was removed.Challenge,3 weeks after intradermal treatment, with 0.5 mL of the test substance (50% solution = 20% active ingredient ) on the left flank and 0.5 mL vehicle (deionised water) on the right flank. After challenge with 50% solution (20% active ingredient), there was 1 animal in the test group with slightly erythema at 24h evaluation. At 48h after challenge none of the 20 animals of the test group had any reaction. In the control group after 24 and 48h there were no reactions on the test site or the control site. In 19 of the 20 test animals no reaction was observed, thus the test substance (50% solution or 20% active ingredient) can be classified as “probably not sensitising”. The significant irritation in 1 test animal can still be assigned to chance.
- In a skin sensitisation study with read-across substance 'Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite' according to the method of Magnusson and Kligman in guinea pigs, the test item containing 39.80% active ingredient was tested for its sensitising properties (Haferkorn, 2013d).A 0.5% suspension in aqua ad iniectabilia chosen for the 1st (intracutaneous) induction stage revealed a discrete or patchy erythema in all 10 animals 24 and 48 hours after administration. 2 mL of a 25% suspension of the test item in aqua ad iniectabilia/animal chosen for the 2nd (topical) induction stage revealed a moderate and confluent erythema 48 hours and a discrete or patchy erythema 72 hours after start of exposure in all 10 animals. The challenge with 2 mL of a 10% suspension of the test item in aqua ad iniectabilia /animal - the maximum non-irritating concentration - revealed no skin irritation in any animal and, thus, the test item had no sensitising properties. The vehicle control revealed no skin reactions. Animals of the same strain treated with α-hexyl cinnamic aldehyde in sesame oil exhibited a sensitising reaction in all animals in form of a moderate and confluent erythema (grade 2). Behaviour of the animals remained unchanged. Body weights were not influenced. The test item containing 39.80% active ingredient, was found to be not sensitising.

 

Human patch testing:

In a skin sensitisation study with read across substance 'Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium salts', a 15 mm patch of the test item containing 35.8% active ingredient (2.5% in petrolatum) was applied to patch sites on the backs or volar forearms of 100 subjects for ten alternate-day 24 hour periods under occlusion (Kligman, 1977). Following a seven-day rest period, 15mm challenge patches of the test item (1% in petrolatum) were applied in the same manner to fresh sites on the backs or volar forearms of all 100 subjects for 24 hours. Challenge sites were read on removal of the patch and 24 hours thereafter, using the 0-4 scale. There were no instances of irritation or sensitisation from this material on the Draize-Shelanski Patch Test. It is unlikely that this test item would present a danger of irritation or sensitisation in normal, intended use.

 

General assessment and conclusion:

- Based on the negative findings in the guinea-pig maximisation and human patch testing with read-across substances, potential for skin sensitisation for registered substance can be excluded.
- Further information supporting the absence of sensitisation potential is also provided in the read across justification for the N2 subgroup, showing that other guinea-pig maximisation was also negative (justification with data matrix separately attached in Section 13).

Migrated from Short description of key information:
Weight-of-evidence on sensitisation potential was available from read across substances in a guinea pig maximisation model and human patch test all indicated there is no sensitisation potential:
-'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts'
- ‘Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite’
-'Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium salts'.
Based on the negative findings in both models for different similar substances, potential for skin sensitisation can be excluded.

Justification for selection of skin sensitisation endpoint:
Most recent study

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on these results and according to the EC Directive (No.93/21/EEC) and CLP (No. 1272/2008 of 16 December 2008), the test substance does not have to be classified and has no obligatory labelling requirement for sensitisation.