Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 19, 2016 - September 26, 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
May 2008, including most recent amendments.
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
August 1998
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147.
Version / remarks:
November 2000, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
dated, 3 November 2015
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
- Name of test material (as cited in study report): di-C16-C18 (evennumbered) alkyl tripropylenetetramine
- Purity: 100% (UVCB substance)
- Purity test date: 08 July 2016
- Batch No.: 1330539
- Expiration date of the batch: 04 August 2018
- Appearance: Viscous liquid
- Storage condition of test material: At room temperature container flushed with nitrogen
- pH: 8.5-10.5 at concentration of 1%
- Specific gravity/density: 0.864 at 20°C
Specific details on test material used for the study:
Specific gravity/density: 0.864 at 20°C
No correction was made for the purity/composition of the test item. Adjustment was made for specific gravity of the test item.

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 10 weeks old)
- Weight at study initiation: 192-194 gram.
- Housing: Individually housed in labeled Makrolon cages containing sterilized sawdust as bedding material and paper as cage-enrichment.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 19 September 2016 to 26 September 2016

Administration / exposure

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
One day before exposure (Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.

The test item was applied on an area of approx. 10% of the total body surface, i.e. approx. 18 cm² for females. The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch, successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages.

Frequency: Single dosage, on Day 1.

REMOVAL OF TEST SUBSTANCE
- Washing: Following application, dressings were removed and the skin cleaned of residual test substance using tap water.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.31 mL/kg bw

Duration of exposure:
24 hours.
Doses:
2000 mg/kg bw (2.31 mL/kg bw)

No. of animals per sex per dose:
3 females/dose
Control animals:
no
Details on study design:
Duration of observation period following administration: 7 days (the study was terminated at day 7 due to severe skin reactions)
- Frequency of observations and weighing:
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration) and at death.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 7.
- Necropsy : The animals were sacrificed by an oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
- Other examinations performed: none.
Statistics:
None.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Based on:
test mat.
Remarks on result:
not determinable
Remarks:
Study was terminated before completion of the observation period and number of animals required due to severe skin reactions.
Mortality:
No mortality occured until day 7 when animals were sacrificed for humane reasons.
Clinical signs:
Days 2, 3 and/or 7: Hunched posture, ptosis and piloerection on Days 2, 3 and/or 7
Day 7: Restless behaviour
Body weight:
The changes noted in body weight gain were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
Severe skin reactions were noted for all animals. The skin reactions were visible from removal of the bandage onwards but worsened until day 7 when it was decided to sacrifice the animals for humane reasons and terminated the study.

Day 2: Erythema and grey discoloration of the treated skin
From Day 4 onwards: Thickened areas, scales and/or scabs
From Day 5 onwards: Dry skin
Day 7: Visible wound, animals were scratching the treated skin and were hypersensitive to touch.

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
In an acute dermal toxicity study with rats, performed according to OECD/EC test guidelines and in compliance with GLP principles, no dermal LD50 value for di-C16-C18 (evennumbered) alkyl tripropylenetetramine could be established since the study was terminated before completion of the observation period and number of animals required due to severe skin reactions.
Executive summary:

An acute dermal toxicity study was performed according to OECD/EC guidelines (OECD 402) and in compliance with GLP principles. Di-C16-C18 (evennumbered) alkyl tripropylenetetramine was administered in a step-wise approach to three female Wistar rats by a single dermal application at 2000 mg/kg body weight for 24 hours. Severe skin reactions were noted for all three animals. The skin reactions were visible from removal of the bandage onwards but worsened until day 7 when it was decided to sacrifice the animals for humane reasons and terminate the study. No further animals were treated. Observed clinical signs included hunched posture, ptosis and piloerection on Days 2, 3 and/or 7 and restless behaviour on Day 7. Local skin effects included erythema and grey discoloration on Day 2, thickened areas, scales and/or scabs from Day 4 onwards and dry skin from Day 5 onwards. On Day 7 a wound was visible and animals were hypersensitive to touch and scratching the treated skin. No dermal LD50 value for di-C16-C18 (evennumbered) alkyl tripropylenetetramine in Wistar rats could be established since the study was terminated before completion of the observation period and number of animals required.