Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 August 2013 - 23 August 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD guidelines and GLP.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Name: PREPOLYMER D
Chemical Name: Reaction mass of Bis (2-chloroethoxy)methane and 1,15-dichloro-3,5,8,11,13-pentaoxa-pentadecane and 1-(2-chloroethoxy)-2-(2-chloroethoxymethoxy)ethane
Batch/Lot Number: 030613
Purity: 100% (multi constituent substance)
Appearance: slightly brownish liquid

Test animals

Species:
rat
Strain:
other: RccHan:WIST
Sex:
female
Details on test animals and environmental conditions:
Species and strain: RccHan:WIST rats
Source: Harlan Laboratories S.r.l.
S.Pietro al Natisone (UD), Zona Industriale Azzida, 57 33040, Italy
Hygienic level at arrival: SPF
Hygienic level during
the study: Standard housing conditions
Number of animals: 12 animals,
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, ~9 weeks old
Date of receipt: 24 July 2013
Body weight at treatment: 168 – 200 g
Acclimation period: 13, 14, 15, 16 days

Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 522/5
Housing: 3 animals / group
Cage type: Type II polypropylene/polycarbonate
Bedding: Lignocel Bedding for Laboratory Animals was available to animals during the study.
A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 35 - 76 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.
The temperature and relative humidity were recorded twice daily during the study.

Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
A single oral gavage administration was followed by a fourteen-day observation period. On the day before treatment, the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment.
Doses:
Initially, three females assigned to (Group 1) were treated at a dose level of 2000 mg/kg bw. The test item caused mortality in this group (3/3). Therefore, a second group (Group 2) was treated at a dose level of 300 mg/kg bw.
The second group of three females (Group 2) was treated at a dose level of 300 mg/kg bw. The test item caused mortality in this group (2/3). Therefore, a third group (Group 3) was treated at a dose level of 50 mg/kg bw.
The third group of three females (Group 3) was treated at a dose level of 50 mg/kg bw. The test item did not cause mortality in this group; therefore a confirmatory group of three females (Group 4) was treated at the same dose level. The test item did not cause mortality in the confirmatory group, so no further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.
No. of animals per sex per dose:
3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on the surviving animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly after. Moreover, the body weight of found dead animals was recorded at necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: None
Statistics:
None

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 30 - <= 300 mg/kg bw
Based on:
test mat.
Mortality:
PREPOLYMER D caused mortality at the dose level of 2000 mg/kg bw (3/3) three to six hours after administration and at the dose level of 300 mg/kg bw (2/3) 1 day after administration.
Clinical signs:
Treatment with PREPOLYMER D at the dose level of 2000 mg/kg bw caused decreased activity (3/3), hunched back (1/3), prone position (3/3), and death (3/3).
Treatment with PREPOLYMER D at dose level 300 mg/kg bw caused decreased activity (3/3), excessive digging (2/3), hunched back (3/3), prone position (1/3), piloerection (3/3), and death (2/3).
Treatment with PREPOLYMER D at dose level 50 mg/kg bw caused faeces liquid (3/6).
Body weight:
Body weight gains of PREPOLYMER D treated animals during the study showed no indication of a treatment-related effect on the surviving animals at the dose level of 300 and 50 mg/kg bw.
Gross pathology:
Diffuse red discoloration of the glandular stomach observed in 3/3 found dead animals dosed at 2000 mg/kg bw was considered to be associated with administration of the test item.
Other changes such as dark/red discoloration of the non-collapsed/collapsed lungs, foamy white material in the trachea or red liquid at the perinasal fur were regarded as agonal or post mortem dosed at 2000 and 300 mg/kg bw.
No macroscopic observations were seen in animals dosed at 50 mg/kg bw and terminated on Day 14.
Also, no gross changes were noted in one surviving rat dosed at 300 mg/kg bw and necropsied on Day 14.

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category III
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
PREPOLYMER D was found to be between 50 and 300 mg/kg bw in female RccHan:WIST rats.
Executive summary:

The single-dose oral toxicity of the test substance was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris) in RccHan:WIST rats.

The test substance was administered in PEG 400 at concentrations of 200, 30 and 5 mg/mL with a dosing volume of 10 mL/kg bw.

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment.

Dose at 2000 mg/kg bw (Group 1):

Initially, three females assigned to (Group 1) were treated at a dose level of 2000 mg/kg bw. The test item caused mortality in this group (3/3). Therefore, a second group (Group 2) was treated at a dose level of 300 mg/kg bw.

Dose at 300 mg/kg bw (Group 2):

The second group of three females (Group 2) was treated at a dose level of 300 mg/kg bw. The test item caused mortality in this group (2/3). Therefore, a third group (Group 3) was treated at a dose level of 50 mg/kg bw.

Dose at 50 mg/kg bw (Group 3 and Group 4):

The third group of three females (Group 3) was treated at a dose level of 50 mg/kg bw. The test item did not cause mortality in this group; therefore a confirmatory group of three females (Group 4) was treated at the same dose level. The test item did not cause mortality in the confirmatory group, so no further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.

Clinical observations were performed on the surviving animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

Results

Mortality

The test substance caused mortality at the dose level of 2000 mg/kg bw (3/3) and at the dose level of 300 mg/kg bw (2/3).

Clinical observations

Treatment at the dose level of 2000 mg/kg bw caused decreased activity (3/3), hunched back (1/3), prone position (3/3), and death (3/3).

Treatment at dose level 300 mg/kg bw caused decreased activity (3/3), excessive digging (2/3), hunched back (3/3), prone position (1/3), piloerection (3/3), and death (2/3).

Treatment at dose level 50 mg/kg bw caused faeces liquid (3/6).

Body weight and body weight gain

Body weight gains of treated animals during the study showed no indication of a treatment-related effect on the surviving animals at the dose level of 300 and 50 mg/kg bw.

Macroscopic Findings

Diffuse red discoloration of the glandular stomach observed in 3/3 found dead animals dosed at 2000 mg/kg bw was considered to be associated with administration of the test item.

Other changes such as dark/red discoloration of the non-collapsed/collapsed lungs, foamy white material in the trachea or red liquid at the perinasal fur were regarded as agonal or post mortem dosed at 2000 and 300 mg/kg bw.

No macroscopic observations were seen in animals dosed at 50 mg/kg bw and terminated on Day 14.

Also, no gross changes were noted in one surviving rat dosed at 300 mg/kg bw and necropsied on Day 14.

Conclusion:

The test substance was found to be between 50 and 300 mg/kg bw in female RccHan:WIST rats.