Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 July 2012 to 21 August 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Well documented GLP study performed according to OECD Guideline 402 and EU Method B3 with some minor deviations. These minor deviations were on the temperature and relative humidity ranges and the age of animals at the start of the study. No impact on the outcome of the study was expected.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
temperature and relative humidity ranges were broader than specified by the guideline, no impact on the outcome of the study; age range 7 to 9 weeks instead of 8-24 weeks, no impact on the outcome or integrity of the study
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
temperature and relative humidity ranges were broader than specified by the guideline, no impact on the outcome of the study; age range 7 to 9 weeks instead of 8-24 weeks, no impact on the outcome or integrity of the study
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): JEFFAMINE RFD-270
- Substance type: Clear colorless liquid
- Physical state: liquid
- Analytical purity: 92.08% primary amine
- Composition of test material, percentage of components: 0.12% water
- Purity test date: 27 July 2010
- Lot/batch No.: 880289
- Storage condition of test material: Room temperature, 15 to 30°C

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan
- Age at study initiation: 7 to 10 weeks
- Weight at study initiation: 184 to 300 grams
- Fasting period before study: no data
- Housing: Animals were group housed by sex upon receipt and individually housed upon assignment to study.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Study animals were acclimated to their housing for a minimum of five days prior to dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 25°C
- Humidity (%): 30 to 84%
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: > = 10%
- Type of wrap if used: The test article was applied on an intact skin site for each animal, covered with a gauze patch/dental dam, wrapped with an elastic bandage and secured with non-irritating tape for 24 (± 0.5) hours.

REMOVAL OF TEST SUBSTANCE
Post-exposure, the site was unwrapped and wiped appropriately (with gauze and water) to remove residual test article.

TEST MATERIAL
Dosage levels were calculated on a mg/kg basis for each individual animal based on body weight. For the liquid test article, the dose was adjusted for test article relative density (1.04 g/mL).

Duration of exposure:
24 ± 0.5 hours
Doses:
One group of ten animals (5/sex) received the test article at 2000 mg/kg (2.1 mL/kg). A full definitive test was performed and three groups of ten (5/sex) received the test article at 50 (5 mL/kg), 200 (5 mL/kg), and 1000 mg/kg (1 mL/kg).
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Morbidity: once daily
Clinical observations: Clinical observations were recorded immediately after unwrap and daily thereafter through day 15. At the time of clinical observations, the dose site was also evaluated for dermal irritation. When present, dermal irritation was scored and recorded using descriptive scoring.
Body weight: Animals were weighed prior to dosing on Day 1 and on days 8 and 15 or upon death.
- Necropsy of survivors performed: yes: all surviving animals were euthanized by CO2 asphyxiation following final observation.
The terminal necropsy included examination of:
- the external body surface
- all orifices
- the thoracic, abdominal and pelvic cavities and their contents
Statistics:
Body weights were summarized using descriptive statistics (mean and standard deviation).

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 000 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed at 50 or 200 mg/kg. Nine of ten animals were found dead at 2000 mg/kg and one of ten were found dead at 1000 mg/kg.
Clinical signs:
All animals at 50 mg/kg were observed to be clinically normal without signs of dermal irritation throughout the study. At 2000 mg/kg, all ten animals exhibited signs of irritation and necrosis at the application site by day 2 with clinical observations. The one remaining animal in this group continued with irritation and necrosis and was clinically normal at day 14. At 200 mg/kg, irritation was observed in four animals starting on day 2 with full resolution by day 6. Clinical observations were present in one animal at 200 mg/kg on day 2 only. At 1000 mg/kg, irritation was observed starting on day 2 in eight animals with necrosis beginning in three of ten animals on day 4. Irritation almost completely resolved in the remaining animals by day 15. Clinical observations were present in five animals at 1000 mg/kg on day 2 only.
Body weight:
Effects in body weight were seen in nine of ten animals in the 2000 mg/kg group with an average weight loss of 14 grams. One animal in the 2000 mg/kg group exhibited a weight gain of 11 grams. No effect in body weight was seen during the study in the 50 mg/kg group with all animals gaining weight by day 15. One animal maintained its weight between day 1 and day 8 and one animal lost weight between day 1 and day 8. No significant effects in body weight were seen during the study in the 200 mg/kg group with nine of ten animals gaining weight between days 1, 8 and 15. One animal in this group lost weight between day 1 and day 8, but overall gained weight by day 15. Effects in body weight were observed in two of ten animals in the 1000 mg/kg group. One animal suffered a weight loss of 8 grams and one animal suffered an overall weight loss of 4 grams (by day 15). The remaining animals gained weight between days 1, 8, and 15.
Gross pathology:
No visible lesions were observed in the animals at terminal necropsy. Necropsy of found dead animals revealed: fluid filled stomachs in four of eight animals found dead in the 2000 mg/kg group on day 3 as well as dark red intestines in one animal found dead on day 4. Necropsy of the found dead animal in the 1000 mg/kg group on day 3 revealed dark red intestines.

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based upon the results of the acute dermal toxicity study in rats with JEFFAMINE RFD-270, the estimated LD50 was considered to be greater than 1000 mg/kg but less than 2000 mg/kg based on the results of the preliminary test (9 of 10 rats were found dead). Therefore, based on the criteria of the CLP Regulation, the substance is considered to be a category 4 acute dermal toxicant.