Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-09-15 to 2011-10-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Commission Regulation (EC) No 440/2008
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
1-butylpyrrolidin-2-one
EC Number:
222-437-8
EC Name:
1-butylpyrrolidin-2-one
Cas Number:
3470-98-2
Molecular formula:
C8H15NO
IUPAC Name:
1-butylpyrrolidin-2-one
Test material form:
other: solution
Details on test material:
- Name of test material (as cited in study report): N -Butylpyrrolidone
- Substance type: organic
- Physical state: clear liquid
- Analytical purity: 99.8w%
- Lot/batch No.: G7844
- Expiration date of the lot/batch: 2012-02-29
- Stability under test conditions: Unknown
- Storage condition of test material: at room temperature (20 ± 5°C, provided by Harlan Laboratories Ltd.) protected from light.

Test animals

Species:
rat
Strain:
other: RccHan: WIST(SPF)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories B.V., Horst, The Netherlands.
- Age at study initiation: 10 weeks (when treated).
- Weight at study initiation: 172.2 g - 188.7 g (when treated).
- Fasting period before study: yes, overnight prior to treatment (18 to 19 hours) and approximately 3 - 4 hours post dose.
- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ("Lignocel" J. Rettenmaier & Söhne GmbH CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) including paper enrichment Enviro Dri (Lillico) (batch no. 75).
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 44/11 (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) ad libitum (except for the overnight fasting period prior to treatment and approximately 3 - 4 hours post dose).
- Water (e.g. ad libitum): Community tap-water from Itingen was available ad libitum in water bottles.
- Acclimation period: 5 days; under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
From: 2011-09-15 To: 2011-09-20 (group 1: the 2000 mg/kg bw dose group);
From: 2011-09-22 To: 2011-10-06 (group 2: the 300 mg/kg bw dose group);
From: 2011-10-04 To: 2011-10-18 (group 3: the 300 mg/kg bw dose group).

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.2 g/mL and 0.03 g/mL.
- Amount of vehicle (if gavage): 10 mL/kg body weight.
- Justification for choice of vehicle: A 20% (weight/weight) mixture with bi-distilled water formed a clear solution suitable for oral gavage application.
- Purity: bi-distilled water

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight.

DOSAGE PREPARATION:
The dose formulations were prepared shortly before each dosing occasion using a magnetic stirrer.
The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight : volume).
Homogeneity of the test item in the vehicle was maintained until and during administration using a magnetic stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not reported
Doses:
single doses of 300 and 2000 mg/kg bw
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
The animals were observed for viability, mortality and clinical signs (daily during the acclimatization period and at approximately 30 minutes and 1, 2, 3 and 5 hours after administration on test day 1 (in common with the clinical signs) and twice daily during days 2 - 15).
Body weight were recorded on test days 1 (prior to administration), 8 and 15.
- Necropsy of survivors performed: yes
All animals which died spontaneously during the observation period were necropsied. All surviving animals were killed at the end of the observation period by carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. An external examination and opening of the abdominal and thoracic cavities for examinations of major organs were performed. The appearance of any macroscopic abnormalities was recorded. No organs or tissues were retained.
- Other examinations performed: clinical signs, body weight.
Statistics:
No statistical analysis was used.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
act. ingr.
Mortality:
All six animals treated with 300 mg/kg of N-Butylpyrrolidone survived until the end of the study period. Two females treated with 2000 mg/kg of the test item died after the administration and one of them was sacrificed in extremis also on the day of treatment.
Clinical signs:
The females treated with 2000 mg/kg showed shortly after treatment moderate convulsions, tachypnea, prostration, clear lacrimation in both eyes and were found unconscious and two of them were found dead after treatment. The females treated with 300 mg/kg of the test item had a slight to moderately decreased activity and slightly ruffled fur. The three further females treated with 300 mg/kg had no clinical signs. No clinical signs were observed during days two to 15.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necropsy.
Other findings:
No other findings were observed.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The median lethal dose of N-Butylpyrrolidone after single oral administration to female rats, observed over a period of 14 days is: 300 mg/kg body weight
Executive summary:

The study was conducted to assess the acute toxicity of N-Butylpyrrolidone in rats. Three female RccHan: WIST (SPF) rats, were treated with N-Butylpyrrolidone by oral gavage administration at a dosage of 2000 mg/kg body weight and two further groups of three females each were treated with 300 mg/kg body weight of the test item. The test item was formulated in purified water at a concentration of 0.2 g/mL and 0.03 g/mL and administered at a dosing volume of 10 mL/kg. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs approximately 30 minutes after treatment and again at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 - 15. Mortality and viability was recorded together with 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. All six animals treated with 300 mg/kg of N-Butylpyrrolidone survived until the end of the study period. Two females treated with 2000 mg/kg of the test item died after the administration and one of them was sacrificed in extremis also on the day of treatment. The females treated with 2000 mg/kg showed shortly after treatment moderate convulsions, tachypnea, prostration, clear lacrimation in both eyes and were found unconscious and two of them were found dead later. The females treated with 300 mg/kg of the test item had a slight to moderately decreased activity and slightly ruffled fur. The three further females treated with 300 mg/kg had no clinical signs. No clinical signs were observed during days two to 15. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy. The median lethal dose of N-Butylpyrrolidone after single oral administration to female rats, observed over a period of 14 days is: 300 mg/kg body weight < LD50 (female rat) < 2000 mg/kg body weight.