Registration Dossier

Administrative data

Description of key information

Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine was found to be irritating to skin, but not corrosive, in in vitro studies using the EpiDerm™ skin model conducted according to OECD Test Guidelines (Dreher, 2012a and b). In eye irritation studies, while the substance was not irritating in an in vitro BCOP assay, it was irritating to rabbit eyes in a study in vivo.

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation, other
Remarks:
in vitro eye irritation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 January to 12 February 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted according to relevant test guidelines, with no deviations.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
The test animal was one male New Zealand White rabbit, obtained from Harlan UK Ltd., Bicester (HsdIf: NZW strain). The rabbit weighed 3.4 kg on Day -1, and was approximately 20 weeks old at administration. The animal was held in stock under laboratory conditions until the day before administration. A clinical examination and eye examination was performed prior to the study to ensure the animal was suitable for the test procedures. The rabbit was individually housed. Global Diet 2930C (Harlan Teklad, Bicester, UK) was provided ad libitum. Mains water was provided ad libitum via water bottles. Temperature was maintained at 15 to 21°C, humidity at 45%, and light was provided on a 12 hour cycle. There were 15 to 20 air changes per hour.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
Not applicable
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
1
Details on study design:
Both eyes of the rabbit were examined for indications of corneal, iridial or conjunctival damage or irritation on Day-1. After initial visual examination, one drop of 1% aqueous fluorescein solution was instilled into both lower conjunctival sacs, allowed to disperse for thirty seconds and removed from the eyes by irrigation with approximately 10 mL water for irrigation jetted gently from a syringe. The corneal surface was illuminated with an ultraviolet source and inspected for areas of absorption of the fluorescing dye that would indicate epithelial damage. Only rabbits with eyes free from damage or irritation were accepted onto study. One rabbit was dosed initially. The lower eyelid of the left eye was gently pulled aware from the eyeball, and of 0.1 mL test material was instilled into the conjunctival sac. After instillation the eyelids were held closed for a few seconds to prevent loss of the test material. The right eye remained untreated to serve as a control. The eyes were not washed for 24 hours after dosing. Serious ocular changes were observed in the first rabbit, therefore no further rabbits were dosed.
The rabbit was observed twice daily for general health and mortality. The rabbit was weighed on the day before administration, and following the last observation. Ocular reactions were recorded 30 minutes, 1 hour and 4 hours after treatment, and once on Day 2, 3 and 4 (at approximately 24, 48 and 72 hours post-treatment). Additional observations were performed twice daily up to Day 22 as necessary. Reactions were scored according to the Draize system. Fluorescein solution was used to assess corneal damage at the 24 hour observation. Body weight was recorded the day before dosing and following the last observation. The animal was sacrificed at the end of the observation period, necropsy was not performed.
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 hours
Score:
1
Max. score:
1
Reversibility:
not reversible
Remarks:
within 21 days
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 hours
Score:
2
Max. score:
2
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 hours
Score:
2.67
Max. score:
2.67
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 hours
Score:
3.33
Max. score:
3.33
Reversibility:
not fully reversible within: 21 days
Irritant / corrosive response data:
Easily discernible translucent areas of corneal opacity were noted in the treated eye from 24 hours after instillation until Day 6. Scattered or diffuse areas of corneal opacity were noted from Day 7 to the end of the observation period. Iridial inflammation was noted from 30 minutes after instillation and persisted until the end of the observation period. Moderate conjunctival irritation was noted 30 minutes and 1 hour after instillation, with severe conjunctival irritation noted from 4 hours after instillation to Day 6. Moderate conjunctival irritation was noted on Day 7 and persisted until the end of the observation period.
Other effects:
No observations indicative of systemic toxicity or ill health were noted for any rabbits during the course of the study. On Day 7 a veterinary examination was carried out using a slip lamp on the left eye of the animal. The following findings were noted: iritis, scleritis, conjunctivitis, panus starting at edge of cornea and corneal oedema in ventral area, some fluorescein stain uptake dorsally (small spots) and more generally over area (ventrally) affected by oedema. There were no deep ulcerative lesions.

Table 1. Individual ocular response in one rabbit following administration of TOFA_DimerFA_TETA_PAA

Time after treatment

Individual ocular response – male rabbit

Cornea

Iris

Conjunctivae

Opacity

Area

Redness

Chemosis

Discharge

30 mins

0

0

1

2

3

2

1 hr

0

0

1

2

3

2

4 hrs

0

0

1

2

3

3

24 hrs*

2

4

1

2

3

3

48 hrs*

2

3

1

3

4

3

72 hrs*

2

2

1

3

3

3

Day 5*

2

2

1

3

2

3

Day 6*

2

2

1

3

2

3

Day 7*

1

2

1

2

1

3

Day 8*

1

2

1

2

1

2

Day 9*

1

1

1

2

1

2

Day 10*

1

1

1

2

2

3

Day 11*

1

1

1

2

2

3

Day 12*

1

1

1

1

1

3

Day 13*

1

1

1

1

1

3

Day 14*

1

1

1

1

1

3

Day 15*

1

1

1

1

1

3

Day 16*

1

1

1

1

1

3

Day 17*

1

1

1

1

1

3

Day 18*

1

1

1

1

1

3

Day 19*

1

1

1

1

1

2

Day 20*

1

1

1

1

1

2

Day 21*

1

1

1

1

1

2

Day 22*

1

1

1

1

1

2

* = fluorescein applied to cornea

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Effects on the cornea, iris and conjunctiva did not reverse within the 21 - day observation period therefore the test material was considered to be corrosive.
Executive summary:

The eye irritant potential of TOFA_DimerFA_TETA_PAA was evaluated in a single, male New Zealand White rabbit according to OECD 405. The undiluted test material (0.1 mL) was instilled into the left conjunctival sac of the rabbit on Day 1, the right eye remained untreated to serve as the control. Ocular reactions were assessed according to the Draize scoring system for 21 days after administration. Instillation of the test material produced easily discernible translucent areas of corneal opacity, iridial inflammation and severe conjunctival irritation. The effects on the cornea, iris and conjunctiva did not reverse within the 21 - day observation period therefore the test material was considered to be corrosive.

Based on the results of this study, the test material is classified as causing irreversible effects on the eye (Category 1) according to Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion

The skin corrosion potential of Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine was investigated in an in vitro EpiDerm™ study, conducted in accordance with GLP, OECD Test Guideline 431 and EU Method B.40 (Dreher, 2012a). Duplicate EpiDerm™ inserts were treated with the test article, distilled water (negative control) and 8N potassium hydroxide (positive control) for 3 minutes and for 1 hour. At the end of the treatment period, the tissues were washed with phosphate buffered saline (PBS) and cell viability was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The skin corrosivity potential was assessed based on the remaining cell viability obtained after test material treatment at the two treatment times. Skin viability after a three minutes or one hour of exposure to the test article was 61% and 35%, respectively. Skin viability after a three minutes or one hour of exposure to the positive control article was 22% and 11%, respectively, demonstrating appropriate performance of the assay. The test article was not corrosive to skin in the in vitro skin model; EpiDerm™.

The skin irritation potential of Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine was investigated in an in vitro EpiDerm™ study, conducted in accordance with GLP, OECD Test Guideline 439 and EU Method B.46 (Dreher, 2012b). In the study, EpiDerm™ SIT (EPI-200) inserts were treated with test article, negative control (phosphate buffered saline (PBS)) and positive control (5% w/v sodium dodecyl sulphate (SDS)) for 60 minutes. At the end of the treatment period, the tissues were washed with PBS and cell viability was assessed using the MTT assay. The skin irritation potential was assessed according to the remaining cell viability obtained after test article treatment. The group mean viabilities were: 5.6% for the test article, 100% for the negative control and 10.7 % for the positive control. The test article was identified as a skin irritant in the in vitro skin model EpiDerm™ SIT (EPI-200) and met the criteria for classification as a Category 2 Skin Irritant according to Regulation (EC) No 1272/2008.

Eye irritation

The potential for Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine to cause corrosion or severe irritation to the eyes was evaluated in excised bovine corneas using the bovine corneal opacity and permeability (BCOP) assay, conducted according to OECD Test Guideline 437 and GLP (Dreher, 2013b). In the study, a volume of 750 µL test material was applied to three separate corneas, followed by a 10 minute incubation period at 32°C ± 1°C. Following incubation, each cornea was washed with media containing phenol red, followed by media without phenol red. The opacities were then measured and the anterior chamber was emptied. To evaluate permeability, sodium fluorescein solution was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following incubation, the media in the posterior chamber was removed and three 350 µL aliquots of the media (per cornea) were analysed for optical density at 490 nm (OD490). The opacity and permeability measurements were used to calculate an In Vitro Irritancy Score (IVIS). The calculated IVIS for the test material was 31.90. As this was below the trigger value of 55.1, the test material was not considered to be corrosive or severely irritating to the eye. However, a further confirmatory test was conducted in an in vivo study.

The potential for Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine to cause eye irritation in vivo was investigated in an eye irritation study in the rabbit conducted according to OECD Test Guideline 405 and GLP (Dreher, 2013c). In the study, undiluted test article (0.1 mL) was instilled into one conjunctival sac of a New Zealand White rabbit on Day 1. Occular reactions were assessed for 21 days after treatment. Instillation of the test article produced easily discernible translucent areas of corneal opacity, iridial inflammation and severe conjuctival irritation. The effects on the cornea, iris and conjunctiva did not reverse within the 21 day observation period. The substance was therefore found to be corrosive to the eye in rabbits.


Justification for selection of skin irritation / corrosion endpoint:
Reliable study providing data from a guideline compliant study. Under the conditions of this study, the test substance was determined to be irritating to skin.

Justification for selection of eye irritation endpoint:
Sole study; guideline and GLP compliant. Under the conditions of this study, the test article was found to be severely irritating to the eye.

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: highly irritating

Justification for classification or non-classification

Skin irritation/corrosion

Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine was identified as a skin irritant in the in vitro skin model; EpiDerm™ SIT (EPI-200). On this basis, the substance meets the criteria for classification as Category 2, H315 “Causes skin irritation” according to Regulation (EC) No 1272/2008, and as Xi, R38 “Irritating to skin” according to Directive 67/548/EEC.

Eye irritation

Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine was found to cause irreversible effects on the eye in an eye irritation study in the rabbit conducted according to OECD Test Guideline 405. On this basis, the substance meets the criteria for classification as Category 1, H318 “Causes serious eye damage” according to Regulation (EC) No 1272/2008 and Xi, R41 “Risk of serious damage to eyes” according to Directive 67/548/EEC.