Condensation products of triethanolamine with addition products of fatty acids, C18 (unsaturated) alkyl with maleic anhydride

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to internationally accepted technical guidelines and in compliance with GLP in a contract research organization. The study is scientifically valid and the report is fully adequate for assessment, despite some minor restrictions (e.g. due to limited reporting).

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

of 1981

Deviations:

yes

Remarks:

24 h instead of 4 h exposure to a formulation of WS400123. Skin examination 0 h, 48 h and 6 days after patch removal.

Qualifier:

according to guideline

Guideline:

other: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, Division of Pharmacology, FDA

Deviations:

not specified

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Number: 5 animals,
- Average bodyweight: 3.0 kg
- Housing: Individual housing in cages.
- Diet: Commercially available standard laboratory diet for rabbits ( Altromin, Lage, Germany).
- Water: Ad libitum

ENVIRONMENTAL CONDITIONS

Room temperature kept constant at 18±2°C, the relative humidity at 50-60%. 12 hours illumination per day.

Type of coverage:

occlusive

Preparation of test site:

other: abraded versus only shaved

Vehicle:

unchanged (no vehicle)

Remarks:

The test substance was a 50% aquaeous formulation

Controls:

not required

Amount / concentration applied:

0.5 mL of the test material (i.e. formulation containing 49.86% WS400123) was administered to the skin by means of a "2.5 cm2" cellulose patch.*

* Comment by the data submitter:
There is no indication in the report, whether the size of treated skin patches was inadvertently stated as only "2.5 cm2" in reality having been sized 2.5 cm x 2.5 cm (i.e. ca. 6 cm2) or whether, contrarily to the cited OECD 404 test guideline, the size of the treated skin patches was really confined to only 2.5 cm2. Regardless whether the treated skin patch was ca. 6 cm2 or only 2.5 cm2, the study results are considered to be valid for assessment.

Duration of treatment / exposure:

24 hours

Observation period:

7 days (i.e. the final observation time point was 6 days post patch removal)

Number of animals:

5

Details on study design:

TEST SITE PREPARATION

Samples of 0.5 mL of the unchanged test material were applied to the shaved intact and to shaved and then abraded skin of albino rabbits by means of cellulose patches. Patches were fixed with Leukoplast(R) and the whole trunk of each animal was enwrapped by a rubber cloth during the 24 hour treatment period, in order to delay evaporation of substances. From the coverage of the whole animal trunk (patches inclusive) with rubber cloth occlusive treatment was concluded by the data submitter, although the report did not specify whether this coverage was considered to be occlusive or semi-occlusive.

Two untreated skin sites (one of shaved intact skin and one of shaved and then abraded skin) served as controls. Only skin reactions different to controls were graded as positive skin reactions.


TREATMENT PERIOD AND REMOVAL OF TEST SUBSTANCE

Occlusive treatment lasted 24 hours. Then the dressings were removed. There was no mentioning in the report that exposed skin patches were cleaned from residual test substance after treatment.

TIME POINTS OF SKIN EVALUATION:

The treated skin patches were evaluated 24 and 72 hours and 7 days post application (i.e. immediately and 48 hours and 6 days post patch removal). The scoring system for the grading of skin reactions is listed in Table 1 (next field below).

Reporting of irritation results in the present robust study summary has been focused to those attained on intact skin, as currently valid test guidelines do not require treatment of abraded skin.

Irritation parameter:

edema score

Remarks:

as defined in Table 1

Basis:

mean

Remarks:

over 5 animals and 2 time points

Time point:

other: 24 and 72 h post treatment start

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 72 hours post treatment start, i.e. within 48 hours post patch removal

Remarks on result:

other: Administered to abraded skin for 24 hours

Irritation parameter:

erythema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #1

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #2

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #3

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #4

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #5

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #1

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #2

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #3

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #4

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Administered to intact skin for 24 hours

Basis:

animal #5

Time point:

other: 24 h, 72 h, 7 d

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Remarks:

as defined in Table 1

Basis:

mean

Remarks:

over 5 animals and 2 time points

Time point:

other: 24 and 72 h post treatment start

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 72 hours post treatment start, i.e. within 48 hours post patch removal

Remarks on result:

other: Administered to abraded skin for 24 hours

Irritant / corrosive response data:

In each animal, shaved intact skin patches treated with a test material formulation (containing 49.86 % WS400123) for 24 hours were free from erythema, scab or edema formation at any observation time point of the study, whereas on abraded skin patches treated with the same dose erythema and edema, both grade 1, were recorded in each animal at 24 hours post treatment start, i.e. immediately after patch removal. These findings were fully reversible, as by 72 hours after treatment start test material treated abraded skin patches did not differ from control abraded skin patches.

Other effects:

None

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Occlusive administration of an undiluted test material formulation (containing 49.86% WS400123) to shaved, intact skin of five rabbits for 24 hours did not induce any skin reactions during the present study. The same treatment to abraded skin induced only erythema and edema, both grade 1, in each of 5 animals, which were fully reversible within 72 hours post treatment start. In view of the absence of any irritating effects of this formulation on intact skin and the very mild and fully reversible erythema and edema reactions of abraded skin, the present study provides supporting evidence that only 4 h exposure of intact skin to undiluted WS400123 would not induce skin reactions necessitating any labelling regarding skin irritation according to EU classification rules [DIRECTIVE 67/548/EEC and REGULATION (EC) 1272/2008].

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation, other

Remarks:

in vivo study performed prior to change in information requirements

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented and reported study fully adequate for assessment. The study was conducted according to internationally accepted technical guidelines and in compliance with GLP in a recognized contract research organization.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

of 2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

of 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

of 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese MAFF Test Data for Registration of Agricultural Chemicals, Eye Irritation (2-1-5), 12 Nohsan No. 8147, Agricultural Production Bureau, November 24, 2000.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS

- Stock supply of healthy adult rabbits
- Number and Sex: 3 females
- Age on day of dosing (Day 1): 40 or 41 weeks
- Weight prior to dosing (Day 1): Minimum 3.97 kg, maximum 4.09 kg
- Housing: Individual housing in plastic cages with peforated floor
- Environmental enrichment: Small soft white untreated wood blocks
- Diet : Standard laboratory rabbit diet (125 g/animal/day). In addition, dietary supplement of hay
replaced by a supplement of wholemeal bread as from 2 days prior to dose instillation.
- Water: Drinking water, ad libitum
- Acclimation period: 29 or 30 weeks prior to study start under laboratory conditions.

Routine analysis of the batch of diet used for nutrients, possible contaminants and microorganisms and of the drinking water were conducted or available at the testing facility.


ENVIRONMENTAL CONDITIONS

Air conditioned animal room set at:
- Temperature (°C): 18 ± 2°C
- Relative Humidity (%): 40 to 70%
- Photoperiod: 12 hrs artificial light per 24 hrs

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL of undiluted liquid test substance was administered into the conjunctival sac of one eye per rabbit (right eye). The contralateral eye (left eye) remained untreated to serve as a control.

Duration of treatment / exposure:

The eyes were not rinsed after treatment. Therefore, the treatment/exposure period was equivalent to the entire observation period following instillation or lasted until removal of the test material by physiological mechanisms, whichever happened sooner.

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

3 adult female rabbits

Details on study design:

EYE EVALUATION:

Before treatment start both eyes of each animal were investigated to ensure that there was no pre-existing corneal damage, iridial inflammation or conjunctival irritation.

One animal (the sentinel) was initially treated and in the absence of a severe effect the remaining two animals were committed to the study.

Eyes were evaluated in all animals at approximately 1, 24, 48 and 72 hours after test substance instillation adopting the numerical scoring system listed in Table 1 in the field below. Grades attained at 24, 48 and 72 hours after instillation were included in the mean gradings of ocular lesions and used for evaluation of the necessity of eye irritation/corrosion classification. As from 24 h after test substance instillation, staining with fluorescein supported the recognition of corneal lesions in female #3.

Equipment used for eye evaluation: Ophthalmoscope or pencil beam torch

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.4

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Reversibility:

other: Corneal ulceration or opacity were not evident

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Reversibility:

other: Corneal ulceration or opacity were not evident

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

iris score

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

2

Reversibility:

other: Iridic changes were not evident

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

2

Reversibility:

other: Iridic changes were not evident

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.4

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.7

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.9

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Discharge

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.9

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Discharge

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

other: Discharge

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: on one occasion mucoid

Irritation parameter:

other: Discharge

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: at times mucoid

Irritant / corrosive response data:

In one animal (#1) ocular lesions were confined to conjunctival redness at 1, 24 and 48 hours after test substance instillation associated with chemosis and discharge at 1 hour after instillation. All of these findings were grade 1 and fully reversible. These findings were also evident in another animal (#2), but at higher severity (up to grade 2) and persisting slightly longer. In addition to conjunctival redness, chemosis and discharge, each up to grade 2, corneal opacity up to grade 2 and iritis grade 1 were recorded in the third animal (#3) all findings being reversible, as by 14 days after instillation also this animal was entirely free from ocular lesions.

Other effects:

Observation of the animals for defined behavioural criteria led to the conclusion that instillation of the test substance induced practically no initial pain response. Any other signs of systemic toxicity or ill health were not evident.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The outcome of the present in vivo study does not necessitate any labelling regarding eye irritation according to EU classification rules [DIRECTIVE 67/548/EEC and REGULATION (EC) 1272/2008]. The findings noted were fully reversible and their incidence is within the category “not irritating to eyes”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The conduct of an in vitro skin corrosion test (OECD 431) was deemed unnecessary for the following reasons:
1. Absence of any dermal irritation in the local lymph node assay in mice treated with WS400123 at up to and including 100% w/v,

2. Non-irritancy of neat WS400123 in the in vitro EPPISKIN skin irritation study, and of WS400123 formulation in the in vivo skin irritation study.

3. Absence of relevant eye irritation or corrosion in the in vivo eye irritation studies with neat WS400123 or WS400123 formulation,
4. Skin corrosion properties of WS400123 are not to be expected from its pH value (measured to be 7.64 as part of the in vivo eye irritation study with neat WS400123).

Justification for classification or non-classification

In view of the absence of any relevant effects during the skin and eye irritation studies with WS400123, the attained results do not necessitate any classification and labelling regarding skin or eye irritation according to EU classification rules [REGULATION (EC) 1272/2008].