9-Octadecenoic acid (Z)-, reaction products with diethylenetriamine, di-Me sulfate-quaternized

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 October 2007 - 20 November 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18 (even numbered) and C18 unsatd., reaction products with diethylene triamine, di-Me sulfate quaternized

EC Number:

937-237-2

Molecular formula:

Molecular formula cannot be given as substance is a mixture.

IUPAC Name:

Fatty acids, C16-18 (even numbered) and C18 unsatd., reaction products with diethylene triamine, di-Me sulfate quaternized

Test animals

Species:

rat

Strain:

other: HanRcc:WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 22 ± 3 °C and for relative humidity between 30-70% (values above 70% during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

corn oil

Details on dermal exposure:

One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10% of the total body surface.
Only those animals without injury or irritation on the skin were used in the test.
On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
Further dosing (200 mg/kg) was administered to 10 naive animals as follows: a single animal of each sex was treated first. Since moderate local effects were observed in one animal of each sex after the 24-hour exposure and by considering the previous results at 2000 mg/kg, the treatment of the remaining four male and four female animals was postponed for 1 week.
The application volume/kg body weight was 4 mL.
Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed.
Seven animals of the first group treated at 2000 mg/kg were superficially re-shaved on test day 6 to facilitate the reading of the local reactions. The two animals treated first at 200 mg/kg were re-shaved on test days 8, 11 and 15 while the remaining animals treated at 200 mg/kg were re-shaved on test days 4, 8 and 11.

Duration of exposure:

24 hours

Doses:

200, 2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

Twenty HanRcc:WIST (SPF) rats, which consisted of one group of 5 males and 5 females, a second group of 1 male and 1 female and a third group of 4 males and 4 females, were treated with the test substance by dermal application. The first group was treated at the dose of 2000 mg/kg b.w. while the remaining two groups were treated at 200 mg/kg. b.w The test item was prepared in vehicle (corn oil) at a concentration of 0.5 or 0.05 g/mL, respectively and administered at a volume dosage of 4 mL/kg. The application period was 24 hours.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Local signs were noted once daily from test day 2 to 15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

Statistics:

No statistical analysis was used

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Remarks on result:

other: no animals died

Mortality:

Due to the severity of the irritations, all animals treated at 2000 mg/kg were humanely sacrificed on test day 10.

Clinical signs:

other: No sign of systemic toxicity was noted

Gross pathology:

No macroscopic findings were recorded at necropsy apart from severe to moderate skin irritation.

Other findings:

Severe signs of dermal irritation at 2000 mg/kg bw. Mild to moderate symptoms of dermal irriation were observed at 200 mg/kg bw.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

On the basis of the results obtained after a single dermal administration, the dermal LD50 of the substance was determined to be > 2000 mg/kg bw.

Executive summary:

In an acute dermal toxicity study according to OECD guideline 402, 1987 and EU method B.3, 1992, 5 male and 5 female young adult HanRcc: WIST(SPF) rats were dermally exposed to the substance suspended in corn oil for 24 hours under a semi-occlusive dressing to approx. 10 % of body surface area at doses of 2000 or 200 mg/kg bw. Low dose group animals then were observed for 14 days. The high dose animals were euthanized due to severe local effects after ten days. No symptoms of systemic toxicity were observed. The local skin reactions affected the study and prevented the full assessment of the LD50. However, even though the 14 day observation period could not be completed the onset of systemic toxicity should have been apparent on day 10.

 

Dermal LD₅₀           Males > 2000 mg/kg bw                                  

Females > 2000 mg/kg bw

                          Combined > 2000 mg/kg bw

 

No mortality occurred in this test.

No clinical signs or gross pathological findings were observed. No weight gain or slight (< 1 %) weight loss was observed in three females of the high dose group (2000 mg/kg bw) and a slight (0.7 % during the first week) but reversible weight loss in one female of the low dose group (200 mg/kg bw).

The minor body weight loss or absence of body weight gain in a few number of females is generally well associated to the female animals which are more sensitive or body weight-affected than the males after dermal exposure. Therefore, the affected body weight suggested a relationship to the type of application and sex of animals rather than any local (the local findings were comparable in both sexes) or systemic toxicity of the test item.The test item is a palmoil – based partially unsaturated IQAC and a read across from fully saturated tallow fatty acid based IQACs can be done. The latter are well-known to have very little (quantified) dermal penetration and no systemic toxicity. The lack of systemic toxicity has been demonstrated in a 91-day percutaneous study with a tallow based IQAC. The findings of this study consisted of moderate erythema, edema, desquamation and fissuring in the high-dose (27 mg/kg/day) animals. Any indication of systemic toxicity or haematologic changes from 13 weeks of dermal application were not observed. Also, no systemic toxicity was observed in the skin sensitisation study, skin irritation or acute oral toxicity study.