bis(diethylamino)-N,N-diethylmethaniminium chloride

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: OECD 402 (1987) and EPA OPPTS 870.1200 (1998) without definitive study; GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

An actual LD50 was not determined.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

An actual LD50 was not determined.

GLP compliance:

yes

Remarks:

US EPA GLP 40 CFR, Part 792 and OECD GLP

Limit test:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

Male and female New Zealand White rabbits were received from Millbrook Breeding Labs, Amherst, MA. The animals weighed 2.14-2.93 kg and were 12-15 weeks old and were individually housed upon arrival in stainless steel suspended cages. The animals were acclimated for at least 5 days prior to dosing. Water and feed were provided ad libitum. The temperature and humidity were maintained at 68±5°F and 30-70%, respectively. Room lights were on a 12-hour light/dark cycle.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

The application site, not less than 10% of the body surface, was prepared approximately 24 hours prior to dosing by clipping the skin of the trunk free of hair. The test substance was dosed as received from the Sponsor. In the limit test, the test substance was dosed at 2000 mg/kg. In the Range finding Test, 2 rabbits/sex were dosed at 250, 500, 1000, 1500 and 2000 mg/kg. The test substance was introduced under gauze patches (two single layers thick) and applied directly to the skin of 10 animals. The animals were immobilized and the patches were secured in place by wrapping the entire trunk of the animal with an impervious bandage. Test sites were secured to prevent the animals from ingesting the test substance. After the completion of the 24-hour exposure period, the wrapping was removed and the skin was gently wiped and rinsed with water to remove any test substance still remaining.

Duration of exposure:

24 hours

Doses:

Limit Test: 2000 mg/kg
Range finding: 250, 500, 1000, 1500 and 2000 mg/kg

No. of animals per sex per dose:

Range finding: 1/sex; Limit test: 5/sex

Control animals:

no

Details on study design:

Five animals/sex were dosed at 2000 mg/kg (limit test). All animals died at this dose. Therefore, a range-finding test was conducted with 1 rabbit/sex dosed at: 250, 500, 1000, 1500 and 2000 mg/kg. The animals were observed frequently during the first day and then a careful clinical examination was made at least once a day through 14 days. The test site of each animal was also observed for signs of erythema and edema after the exposure period according to the Draize Scale for Scoring Skin Reactions. Animals were weighed at Day 0 (prior to dose administration), Day 7 and Day 14. Changes in body weights were calculated and recorded. A gross necropsy was performed on all animals whether found dead in extremis (dead no longer than 12 hours) or sacrificed by an injectable barbiturate at the end of the study.

Results and discussion

Preliminary study:

As no animals died at 500 mg/kg, one of two animals died at 1000 mg/kg and all animals died at doses of 1500 mg/kg and above, and as a definitive LD50 was not performed, the LD50 of the test substance following a single topical application would be expected to be within the range of 500 to 1500 mg/kg.

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

>= 500 - <= 1 500 mg/kg bw

Based on:

test mat.

Mortality:

For the limit test, 8/10 animals died within two hours of dose administration. The remaining animals were found dead at the 24-hour time point. For the range finding test, both animals dosed at 2000 and 1500 mg/kg and one female dosed at 1000 mg/kg died within the first 24 hours. The remaining animals (1000 mg/kg male, both male and female animals dosed at 500 mg/kg and 250 mg/kg) survived to study termination.

Clinical signs:

For the limit test, ataxia was observed in 5/10 animals while somnolence was observed in 1/10 animals prior to death. For the range finding test, the male animal dosed at 2000 mg/kg was observed with dyspnea and lethargy prior to death. The surviving male dosed at 1000 mg/kg exhibited signs of lethargy and abnormal posture for the first 2 days of the study and normal clinical findings for the remainder of the study. Animals dosed with 500 mg/kg and 250 mg/kg did not exhibit any abnormal clinical findings.

Body weight:

All animals that survived to study termination gained weight by the end of the study.

Gross pathology:

There were no abnormalities or lesions observed at necropsy for any of the animals.

Other findings:

No irritation at the dose site was observed in the two animals surviving to 24 hours in the limit test. For the range finding test, no irritation was observed at the dose site throughout the study for the two animals dosed at 250 mg/kg. Well-defined erythema was observed in both animals dosed at 500 mg/kg on Day 1, which decreased in severity until resolved in the female at Day 3 and the male at Day 4. The surviving animal dosed at 1000 mg/kg was observed with well-defined erythema on Day 1 and severe erythema on Day 2 which persisted until study termination on Day 14.

Any other information on results incl. tables

Given the results of the preliminary study, a definitive study was not performed.

Applicant's summary and conclusion

Executive summary:

As no animals died at 500 mg/kg, one of two animals died at 1000 mg/kg and all animals died at doses of 1500 mg/kg and above, and as a definitive study was not performed, the LD50 of the test substance following a single topical application would be expected to be within the range of 500 to 1500 mg/kg.