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Description of key information

The acute oral toxicity of Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines is low. The oral LD50 of the substance was determined to be > 2000 mg/kg bw in rats in an acute oral toxicity study (OECD Test Guideline 423) whereas the dermal LD50 of the substance was determined to be > 2000 mg/kg bw in rats in an acute dermal toxicity study (OECD Test Guideline 402). A waiver is proposed in accordance with Column 2 of Annex VIII of the REACH Regulation for acute inhalation toxicity on the basis that acute toxicity data are available for the oral and dermal routes. Inhalation is not predicted to be a significant route of exposure based on the physicochemical properties of the substance.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted according to relevant guidelines.
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
not applicable
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
not applicable
Principles of method if other than guideline:
Not applicable.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan UK Ltd, Bicester.
- Age at study initiation: 8 to 9 weeks old
- Weight at study initiation: The weight variation did not exceed ±20% of the mean weight.
- Fasting period before study: Animals were fasted from the evening of the day prior to dosing until approximately 3 hours after dosing.
- Housing: The animals were housed in groups of up to five during the acclimatisation period. From the day prior to dosing (Day –1), the rats were housed in groups of three in similar cages.
- Diet (e.g. ad libitum): SQC(E) Rat and Mouse Maintenance Diet No 1, was freely available to the animals at all times, except for a period of fasting from the evening of the day prior to dosing (Day -1) until approximately 3 hours after dosing.
- Water (e.g. ad libitum): Mains water was provided, ad libitum, via cage-mounted water bottles.
- Acclimation period: 7 to 9 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 45 to 65%
- Air changes (per hr): 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): Twelve hours light (illuminated fluorescent strips) anad 12 hours dark.

IN-LIFE DATES: From: To:
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw.

DOSAGE PREPARATION: Due to the viscosity of the test article, it had to be diluted in order for it to be dosed. The test article was dispersed in corn oil because the test article did not suspend in purified water. The formulated concentrations were calculated from the selected dose level and the dose volume of 10 mL/kg bw. All formulations were used within two hours of preparation.
The formulations were maintained by multiple inversion or magnetic stirring prior to administration to ensure homogeneity.

CLASS METHOD
- Rationale for the selection of the starting dose: Since there were no data to indicate that deaths may occur at dose levels of less than 2000 mg/kg, the first dose level was 2000 mg/kg bw.
Doses:
Dose level: 2000 mg/kg bw
No. of animals per sex per dose:
3 female animals used per test.
Control animals:
no
Details on study design:
Two groups of fasted animals were treated with 2000 mg/kg bw test material, administered in a dose volume of 10 mL/kg bw. Three animals were in each test group. The treatment of the animals was sequential, with sufficient time between each group to confirm the survival of the previously dosed animals. Individual dose volumes (mL) were calculated using the fased body weights of the rats on the morning of the day of dosing (Day 1) and the dose volume of 10 mL/kg bw.

Treated rats were observed closely for clinical signs of reaction to treatment. Clinical signs were recorded immediately post-dose, at approximately 15 and 30 minutes post-dose, hourly between 1 and 4 hours post-dose (inclusive), twice daily on Days 2, 3 and 4 and once daily from the fifth to last day of the observation period. Individual records of clinical signs were maintained for each treated rat.

All animals were examined at the beginning and end of the working day throughout the acclimatisation and study periods to ensure they were in good health.

Rats were weighed on Day -1 (day before dosing) and on Days 1, 4, 8 and 15.

Rats were killed on Day 15. The necropsy procedure included inspection of external surfaces and orifices, all viscera and tissue within the abdominal, thoracic and cranial cavities, free-hand sectioning of the liver and kidneys and examination of representative sections of mucosal surfaces of the stomach, small and large intestines. No tissue preservation or histopathological assessment of tissues was undertaken.


Statistics:
Not applied.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths following a single oral dose of DimerFA_PEPA_PAA at 2000 mg/kg bw.
Clinical signs:
No clinical signs were observed.
Body weight:
All rats gained weight during the first and second weeks of the observation period.
Gross pathology:
No macroscopic changes were observed for animals killed on Day 15.
Other findings:
No other findings reported.

No additional information.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the acute median lethal oral dose level of the test article, DimerFA_PEPA_PAA, was found to exceed 2000 mg/kg bw.
Executive summary:

A GLP study was conducted to determine the acute oral toxicity of DimerFA_PEPA_PAA in female HsdHan:WIST rats, according to OECD Test Guideline 423 and EU Method B.1 tris.

Two groups of three female fasted rats were given the test article as a single dose on Day 1 by oral gavage at a dose level of 2000 mg/kg bw. The test article was dispersed in corn oil and administered at a dose volume of 10 mL/kg. During the two week observation period, mortality, body weight changes and any adverse clinical signs were recorded. All animals were killed on Day 15 and subsequently underwent a full necropsy.

There were no deaths following a single oral dose of DimerFA_PEPA_PAA at 2000 mg/kg bw. No abnormal clinical signs were observed during the observation period and all rats achieved body weight gains during the first and second weeks of the study. Macroscopic examination of these animals revealed no abnormalities.

Under the conditions of this study, the acute median lethal oral dose level of the test article, DimerFA_PEPA_PAA, was found to exceed 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Klimisch score = 1. Study compliant with current test guidelines and GLP.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 November 2012 to 19 December 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted according to relevant testing guidelines, with no significant deviations.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
The test site in the preliminary study was 5% of total body surface area instead of 10%; this was not considered to have affected the overall integrity of the study
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
The test site in the preliminary study was 5% of total body surface area instead of 10%; this was not considered to have affected the overall integrity of the study
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: HsdHan:WIST
Sex:
male/female
Details on test animals or test system and environmental conditions:
The animals were 8-10 week old male and female HsdHan:WIST rats, obtained from Harlan UK Ltd., Bicester. Females were nulliparous and non-pregnant. The males weighed 246 to 344 g on Day 1 and females weighed 169 to 219 g. The acclimatisation period was 8 to 16 days.
Rats were housed in same sex groups of up to 5 during the acclimatisation period, and individually from the day prior to dosing. After the Day 3 observation the animals were returned to group housing. The rats were fed SQC(E) Rat and Mouse Maintenance Diet No. 1 (Special Diet Services Ltd., Witham, UK) ad libitum, and mains water was available ad libitum. The animal rooms were maintained at a temperature of 20 to 24°C, relative humidity of 45% to 65% and there were 15 to 20 air changes per hour. Fluorescent lighting was provided on a 12 hour light/dark cycle.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
All hair was removed from the dorsum of each rat on the day before dosing. The test site was an area of at least 10% of the total body surface area, calculated according to the largest animal in each group using the following formula: Surface area (cm²) = K x body weight (g)²/³ (where K = 9).
The test material was spread as uniformly as possible over the test site. The dose volume was 2.08 mL/kg bw, and was calculated from the body weights of the rats on the morning of dosing and the density of the test material. A dense gauze patch was placed over the treated skin and retained in place by an elasticated, open-weave, adhesive compression bandage. This was wrapped securely around the torso of the animal. At the end of the exposure period the dressings were removed and the test site was lightly brushed clean of any solid residues and swabbed with water-moistened cotton wool.
The test material was administered as supplied, and corrections for purity or active content were not made.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5/sex (including preliminary study)
Control animals:
not required
Details on study design:
A preliminary study was conducted with one male and one female rat. Each rat received a single dermal dose of the test substance at 2000 mg/kg bw. Due to a calculation error, the body surface area used was 5% instead of 10%. This was not considered to have affected the overall integrity or outcome of the study as the responses seen in these animals were consistent with those seen in the main study animals. Since no mortalities were observed in the preliminary test, the test material was applied dermally to four male and four female rats at a dose of 2000 mg/kg bw. The rats were observed for 14 days after dosing. Treated rats were observed for clinical signs of toxicity immediately post-dose, at approximately 15 and 30 minutes post-dose, hourly between 1 and 4 hours post-dose (inclusive), twice daily on Days 2, 3 and 4 and once daily from the fifth to the last day of the observation period. Checks for mortality and general health were made at the beginning and end of each working day throughout the acclimatisation period and study period. The second observation on Day 3 for the female main study animals was not performed, but this was not considered to have affected the outcome of the study. Body weights were recorded the day prior to dosing, and on Days 1, 4, 8 and 15. The test site was evaluated for dermal reactions following removal of the dressings on Day 2 and daily therefore for the remainder of the observation period.
Rats were sacrificed on Day 15, and full necropsy was performed and all lesions were recorded. The necropsy procedure included inspection of external surfaces and orifices, the dermal test site, all viscera and tissue within the abdominal, thoracic and cranial cavities, free hand sectioning of the liver and kidneys and examination of representative sections of mucosal surfaces of the stomach and intestinal tract.
Statistics:
Not required.
Preliminary study:
No mortalities occurred in the preliminary study.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities occurred.
Clinical signs:
No clinical signs of toxicity were observed.
Body weight:
All rats gained weight during the first and second weeks of the study, except for one female which showed a slight loss in weight during the first week.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
Very slight erythema was noted at the test site of two females on Day 3 and at the test sites of two males and three females on Day 4. Very slight erythema persisted in one male and three females up to Day 13. Discolouration was noted at the test site of one male on Day 2 and in all males and three females from Day 3 to Day 13. Discolouration was noted in all males on Day 14 and in one male on Day 5. Scabbing was noted at the test sites of one male on Day 3, in two males and three females from Day 4 to Day 7 and in three males and three females from Day 8 to Day 14.
It could not be confirmed if the discolouration of the skin was a treatment-related effect. The test article was described as a brown/yellow liquid but the treatment sites were washed following removal of the bandages and in the majority of cases, the discolouration did not develop until Days 3 or 4.

No further information.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal LD50 was found to exceed 2000 mg/kg bw in rats.
Executive summary:

The acute dermal toxicity of DimerFA_TEPA_PAA was investigated in male and female HsdHan:WIST rats, in a GLP study according to OECD Test Guideline 402. A preliminary study was conducted with one male and one female rat. Following the preliminary study an additional 4 rats per sex were treated. The test material was applied undiluted to the clipped dorsum of the rats at a dose of 2000 mg/kg bw. The test site was covered with a semi-occlusive dressing for 24 hours. Following dressing removal the animals were observed for dermal reactions at the test site, clinical signs of toxicity and mortality for 14 days. All animals were sacrificed at the end of the observation period and subject to full necropsy.

There were no mortalities and no clinical signs of toxicity. Very slight erythema, discolouration and scabbing were noted at the test sites of treated rats, lasting up to Day 15. There were no treatment-related effects on body weight, and no abnormalities were noted at necropsy. Under the conditions of the study, the acute dermal LD50 of DimerFA_TEPA_PAA was found to exceed 2000 mg/kg bw in rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Klimisch score = 1. Study compliant with current test guidelines and GLP.

Additional information

Acute oral toxicity

The acute oral toxicity of Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines is low. The oral LD50of the substance was determined to be > 2000 mg/kg bw in an acute oral toxicity study conducted in rats according to OECD Test Guideline 423 and EU Method B.1 tris (Williams, 2012). 

In the study, two groups of three female fasted rats were given the test article as a single dose on Day 1 by oral gavage at a dose level of 2000 mg/kg bw. The test article was dispersed in corn oil and administered at a dose volume of 10 mL/kg bw. Animals were observed for 14 days following treatment and during this observation period, mortality, abnormal clinical signs and bodyweight were recorded. All animals were killed on Day 15 and subsequently underwent a full necropsy. There were no deaths, and no clinical signs were observed. All rats gained weight during the first and second weeks of the observation period. Macroscopic examination of these animals revealed no abnormalities. Under the conditions of this study, the acute median lethal oral dose level (LD50) of the test article, Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines was found to exceed 2000 mg/kg bw.

Acute dermal toxicity

The acute dermal toxicity of Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines is low. The dermal LD50of the substance was determined to be > 2000 mg/kg bw in an acute dermal toxicity study conducted in rats according to OECD Guideline 402 and EU Method B.3 (Dreher, 2013a).

A preliminary study was conducted using one male and one female rat. Following the preliminary study an additional four rats per sex were treated. The test article was applied undiluted to the clipped dorsum of the rats at a dose of 2000 mg/kg bw. The test site was covered with a semi-occlusive dressing for 24 hours. Following dressing removal the animals were observed for dermal reactions at the test site, clinical signs of toxicity and mortality for 14 days. All animals were sacrificed at the end of the observation period and subject to full necropsy. There were no mortalities and no clinical signs of toxicity. Very slight erythema, discolouration and scabbing were noted at the test sites of treated rats, lasting up to Day 15. There were no treatment-related effects on body weight, and no abnormalities were noted at necropsy. Under the conditions of the study, the acute dermal LD50of the test article, Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines, was found to exceed 2000 mg/kg bw in rats.

Acute inhalation toxicity

A waiver is proposed for acute inhalation toxicity studies in accordance with Column 2 of Annex VIII of the REACH Regulation on the basis that acute toxicity data are available for the oral and the dermal routes. Inhalation is not predicted to be a significant route of exposure based on the physicochemical properties of the substance. The substance is a non-volatile liquid and the use pattern indicates that significant inhalation exposure is unlikely. No additional testing is therefore warranted.


Justification for selection of acute toxicity – oral endpoint
Sole study; guideline and GLP compliant.

Justification for selection of acute toxicity – dermal endpoint
Sole study; guideline and GLP compliant.

Justification for classification or non-classification

Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines has low acute toxicity: the oral LD50and dermal LD50values in rats have both been determined to be > 2000 mg/kg bw/day. The substance does not therefore meet the criteria for classification for acute toxicity according to Regulation 1272/2008/EC or Directive 67/548/EEC.