Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
a short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration
Cross-reference
Reason / purpose:
data waiving: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP; does not fully conform to current guidelines-no individual responses
Justification for type of information:
Refer to section 13 for details on the read-across justification. The study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Remarks:
pre-GLP; QA declaration
Test type:
standard acute method
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Equal numbers of male and female CFY (Sprague-Dawley origin) rats were obtained from Interfauna UK Ltd., Huntingdon, Cambridgeshire, England. They were in a weight range of 89 to 121 g prior to dosing (Day 1) in the main study and approximately four to six weeks of age. All the rats were acclimated to the experimental environment for a period of 5 days prior to the start of the main study.

The rats were allocated to cages within the treatment groups. They were housed in groups by sex in metal cages with wire mesh floors. A standard laboratory rodent diet (Labsure LAD 1) and water were provided ad libitum. The batch of diet used for the study was analysed for certain chemical and microbiological contaminants. Access to food only was prevented overnight prior to and approximately 4 hours after dosing.

The mean daily minimum and maximum temperatures of the animal room were 21°C and 22°C respectively and the mean daily relative humidity value was 47%. The rate of air exchange was maintained at approximately 15 air changes/hour. Lighting was controlled by means of a time switch to 12 hours artificial light in each 24 hour period.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Redicote EN611 was administered, as supplied by the Sponsor, at a volume not exceeding 1.52 mL/kg (S.G. 1.05) in the main study.
Doses:
A trial test was carried out to establish a dosing regimen for the main study using groups of two male and two female rats at three dose levels of 1000, 2500 and 5000 mg/kg bodyweight.

Doses selected for the main study were 640, 1000 and 1600 mg/kg bodyweight.

No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
The appropriate dose volume of the test substance was administered to each rat using a syringe and plastic catheter.

Animals were observed soon after dosing; then at frequent intervals for the remainder of Day 1. On subsequent days the animals were observed once in the morning and again at the end of the experimental day. This latter observation was at approximately 16.30 hours on weekdays or 11.30 hours on Saturday and Sunday. Clinical signs were recorded at each observation.

The animals on the preliminary and main studies were observed for 5 and 14 days respectively, after dosing.

Surviving animals on the main study were killed on Day 15 by cervical dislocation. All animals that died during the study and those killed on Day 15 were subjected to a macroscopic post mortem examination which consisted of opening the abdominal and thoracic cavities. The macroscopic appearance of abnormal organs when present was recorded.
Statistics:
The acute median lethal oral dose (LD50) to male and female rats was calculated using the method of: Finney (1971) Probit Analysis (3rd Edition) Cambridge University Press. Separate LD 50 values for males and females were estimated by undertaking probit analysis on the mortality data by fitting two parallel lines on the data (males only and females only) using the technique described by Finney 1978, Statistical Method in Biological Assay, 3rd Edition, Charles Griffin, London). A chi-squared test was carried out to check that the data did not contain any evidence for non-parallelism.
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
1 145 mg/kg bw
Based on:
test mat.
95% CL:
ca. 871 - ca. 1 543
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
1 164 mg/kg bw
Based on:
test mat.
95% CL:
ca. 893 - ca. 1 506
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 156 mL/kg bw
Based on:
test mat.
95% CL:
ca. 956 - ca. 1 405
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
343 mg/kg bw
Based on:
act. ingr.
Remarks:
Based on 30% of 30-40% active substance
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
349.2 mg/kg bw
Based on:
act. ingr.
Remarks:
Based on 30% of 30-40% active substance
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
347 mg/kg bw
Based on:
act. ingr.
Remarks:
Based on 30% of 30-40% active substance
Mortality:
There were deaths among male and female rats treated at 1000 (3/10) and 1600 mg/kg (9/10) from within one hour of dosing to Day 2.
Clinical signs:
Signs of reaction to treatment observed in all rats shortly after dosing were pilo-erection, abnormal body carriage (hunched posture), abnormal gait (waddling) and increased salivation. These were accompanied by: lethargy and decreased respiration in all animals surviving for more than one hour after treatment at 1000 or 1600 mg/kg, ptosis in one rat dosed at 640 mg/kg and in all rats surviving for more than one hour after treatment at higher doses, pallor of the extremities of one rat dosed at 640 mg/kg and in all animals given higher doses, diarrhoea in all rats treated at 640 mg/kg. Recovery, as judged by external appearance and behaviour, was advanced by Day 3 and complete by Day 5.
Body weight:
Bodyweight losses or no change of bodyweight were recorded for rats that died. Slightly low bodyweight gains were recorded on Day 8 for one male dosed at 1000 mg/kg and on Day 15 for one female treated at 640 mg/kg. Other rats achieved anticipated bodyweight gains throughout the study.
Gross pathology:
Autopsy of rats that died commonly revealed renal pallor. There were no other macroscopic abnormalities.Terminal autopsy findings were normal.

Time and number of deaths

Sex

Dose

(mg/kg)

# deaths

Total #

animals

Day 1

Day 2

Day 3

Day 4

Day

5-15

a     b

a     b

a     b

a     b

a     b

M

640

0

5

M

1000

1

5

1

M

1600

5

5

5

F

640

0

5

F

1000

2

5

        2

F

1600

4

5

4

a = first, b = second observation

Interpretation of results:
other: Category 4 based on CLP criteria
Conclusions:
Under the study conditions, the acute oral LD50 of the substance in rats was determined to be 1145 mg/kg bw (i.e., equivalent to 343 mg a.i./kg bw).
Executive summary:

A study was conducted to determine the acute oral toxicity of the read across substance, N,N,N',N',N''-Pentamethyl-N-C16 -18 (even numbered) and C18 unsat.-alkyl-1,3 -propanediammonium chloride, in Sprague-Dawley rats according to OECD Guideline 401. A preliminary test was conducted using nominal dose levels of 0, 1000, 2500 and 5000 mg/kg bw. Based on the results of the preliminary test, the main test was conducted at nominal doses of 0, 640, 1000 and 1600 mg/kg bw. Groups of five (male and female) Sprague-Dawley rats (5/dose) received the test substance. Mortality was observed in male and female rats treated at 1000 (3/10) and 1600 mg/kg bw (9/10) from one hour of dosing to Day 2. Autopsy of the dead animals revealed only pallor of the kidneys and no other macroscopic abnormalities. Gastro-intestinal haemorrhage was expected, but it is not clear whether the inside of the gastro-intestinal system was examined. Acute oral LD50 in rats was determined to be 1145, 1164 and 1156 mg/kg bw for male and female rats and for combined sex respectively. Under the study conditions, the acute oral LD50 of the substance in rats was determined to be 1145 mg/kg bw (equivalent to 343 mg a.i./kg bw) (Gardner, 1987).

Data source

Materials and methods

Results and discussion

Applicant's summary and conclusion