Registration Dossier

Administrative data

Description of key information

The oral LD50 for the test substance was found to be between 50 and 300 mg/kg bw in female RccHan:WIST rats. The dermal LD50of the test substance was found to be higher than 2000 mg/kg bw in male and female CRL:(WI) rats. Based on the very low vapour pressure of the substance and a pattern of use that does not result in the formation of aerosoles an acute inhalation study was not performed. However, an acute inhalation study is available for the major constituent of the substance, 40% is bis(2-chloroethoxy)methane CAS 111 -91 -1 + homologues. The inhalatory LC50, 4h value for this substance is within the range of 0.05 – 0.5 mg/L.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
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.
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
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.
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
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.
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.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Quality of whole database:
One K1 study is available.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
30 September 2009 - 06 January 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: performed according to OECD test guidelines and GLP compliant
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The registered substance is a reaction mass containing the read across source in an amount of 40%. A lower vapour pressure is found for this constituent than for the reaction mass,. Thus this inhalation study is expected to represent a worst case situation.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source chemical:
bis(2-chloroethoxy)methane; CAS: 111-91-1
Purity:90%

Target chemical:
Reaction mass of 1,15-dichloro-3,5,8,11,13-pentaoxapentadecane and 1-chloro-2-[(2-chloroethoxy)methoxy]ethane and 1-chloro-2-{[2-(2-chloroethoxy)ethoxy]methoxy}ethane/ EC number: 940-783-4

Constituent CAS %
bis(2-chloroethoxy)methane 111-91-1 40
1,15-dichloro-3,5,8,11,13-penta-oxa pentadecane - 25
2-chloroethoxymethyl-2-chloroethoxyethyl-ether 117994-78-2 10
Impurity:
Chloroethanol 107-07-3 0,3

3. ANALOGUE APPROACH JUSTIFICATION

Based on available measured and predicted vapour pressure data and Molecular weights, the target chemical can be expected to represent the chemical most abundant in inhaled vapours, and also be a worst case representative of inhalation toxicity.

4. DATA MATRIX
Multiconstituent substance
Constituent CAS % vp measured, Pa vp predicted (Modified Grain Method), Pa SMILES MW
bis(2-chloroethoxy)methane 111-91-1 40 1,76 18 ClCCOCOCCCl 173
1,15-dichloro-3,5,8,11,13-penta-oxa pentadecane - 25 0,00647 ClCCOCOCCOCCOCOCCCl 291
2-chloroethoxymethyl-2-chloroethoxyethyl-ether 117994-78-2 10 1,17 ClCCOCOCCOCCCl 217
Impurity:
Chloroethanol 107-07-3 0,3 957 535 OCCCl 80



Monoconstituent substance
Constituent % vp measured, Pa vp predicted (Modified Grain Method), Pa SMILES MW
bis(2-chloroethoxy)methane 111-91-1 90 1,76 18 ClCCOCOCCCl 173
Impurity:
Chloroethanol 107-07-3 0,3 957 535 OCCCl 80
Bis(2-chloroethoxy)dimethylether 73131-00-7 1 3,28 ClCCOCOCOCCCl 203
2-chloroethoxymethyl-2-chloroethoxyethyl-ether 117994-78-2 7 1,17 ClCCOCOCCOCCCl 217
1,16-dichloro-3,5,8,11,14-pentaoxa hexadecane 0,2 0,00275 ClCCOCOCCOCCOCCOCCCl 305
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Species Rat: Crl:WI(Han) (outbred, SPF-Quality)
Recognised by international guidelines as the recommended test system (e.g. OECD, EC).
Source: Charles River Deutschland, Sulzfeld, Germany.
Number of animals 12; 3 males and 3 females (females were nulliparous and non-pregnant) per exposure level. Two exposure levels were used.
Age and body weight Young adult animals were selected (approximately 11 weeks old).
Animals used within the study were of approximately the same age and body weight variation did not exceed +/- 20% of the sex mean.
Identification Earmark
Health inspection A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health.

Conditions
Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0C (actual range: 18.4 – 21.8C), a relative humidity of 40-70% (actual range: 31 - 76%) and 12 hours artificial fluorescent light and 12 hours darkness per day.
During exposure the temperature and relative humidity were between 19.2 to 21.3oC (mean 20.6 ± 0.6 oC) and 10 to 14% (mean 11 ± 1%) respectively.

Accommodation
Group housing of 3 animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
After exposure on Day 1, the animals exposed at 0.55 mg/L were housed in stainless steel wire mesh cages to prevent suffocation in case of bad health condition and in order to recover. At the end of Day 1 the animals were housed as described above.

Acclimatisation period was at least 5 days before start of treatment under laboratory conditions. Animals were housed with maximally 5 animals per cage per sex as described above.

Diet
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance.

Water
Free access to tap water except during exposure to the test substance.

Results of analysis for each batch of diet (nutrients and contaminants), sawdust, paper and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
Animals were exposed to the test substance via the inhalatory route. For this purpose the animals were placed in restraining tubes, connected to the exposure chamber.
The design of the exposure chamber was based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of three animal sections with eight animal ports each. Each animal port had its own atmosphere inlet and exhaust outlet. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 2.0 L/min, which ensures an adequate oxygen supply to the test animals. The main inlet of the test atmosphere was located at the top section and the main exhaust outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
The placement of the individual animals in the inhalation chamber is shown in figure 2. All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained a slight negative pressure.

Since during method development of the atmosphere generation of 0.55 mg/L the test atmosphere contained droplets, the FTIR readings were compared with the results of samples measured gravimetrically. Gravimetrically a mean of 0.41 mg/L (n=4) was calculated for the aerosol.
By placing a filter (PTFE, 0.2 um) in the FTIR sample line the amount of gaseous phase was measured and a mean of 0.14 mg/L (n=8) was calculated. Since the FTIR readings at development showed a concentration of 0.57 mg/L it was concluded that the both phases were collected and measured correctly by the FTIR and that no gravimetrical measurements were needed for the main study.

Method development for the 0.055 mg/L showed gravimetrically an aerosol concentration of approximately 0.008 mg/L (n=2) at FTIR concentrations of approximately 0.056 mg/L, indicating an aerosol content of approximately 14%.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Infrared spectrophotometer
Duration of exposure:
4 h
Concentrations:
0.55 and 0.055 mg/L
No. of animals per sex per dose:
3 males / 3 females
Control animals:
no
Details on study design:
Target concentrations were based on the cut off values specified in the UN and EC classification guidelines for dusts and mists (0.05, 0.5, 1 or 5 mg/L.) increased with 10% in order to avoid the actual mean concentrations to fall below the cut off values due to experimental variation.

Based on available toxicity data of the test substance and since it is the intention to minimize the number of animals to be used, 0.55 mg/L was selected as starting concentration. Three males and three females were exposed and were found dead on Days 1 (Day of exposure) or day 2. In consultation with the sponsor, a second group of three males and three females were exposed at 0.055 mg/L.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 0.05 - <= 0.5 mg/L air
Exp. duration:
4 h
Mortality:
All animals exposed to 0.55 mg/L were found dead between Days 1 and 2 post-exposure. No mortality occurred at 0.055mg/L.
Clinical signs:
other: During exposure, no clinical signs were noted. After exposure, lethargy, hunched posture, ventral-lateral recumbence, slow breathing and/or piloerection were noted among the animals exposed at 0.55 mg/L. No clinical signs were noted in the animals expose
Body weight:
Overall body weight gain in surviving males and females were within the range expected for rats of this strain and age used in this type of study
Gross pathology:
No treatment abnormalities were found at macroscopic post mortem examination of the animals.

The mean actual concentrations were 0.55 ± 0.07 mg/L and 0.058 ± 0.011 mg/L. The nominal concentrations were 1.895 and 0.99 mg/L and the generation efficiencies (ratio of actual and nominal concentration) were 29 and 6% respectively.

 

During the exposure to 0.55 mg/L, the Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. The MMAD was 3.5mm and 4.1mm respectively and the gsd was 1.6 and 1.5 respectively. The MMAD of the second measurement was just abovethe recommended range of 1 – 4µm. Since one MMAD value was within the range and the other close to the upper limit and since the gsd were appropriate (i.e. between 1.5 and 3), it can be assumed that deposition of particles in the lower respiratory tract had occurred.For the 0.055 mg/L, an attempt was made to determine the MMAD and gsd but due to the very small amount of aerosol present (approximately 0.008 mg/L) the measurement did not show a normal distribution and therefore no MMAD and gsd were calculated.

Interpretation of results:
Category 2 based on GHS criteria
Remarks:
Migrated information
Conclusions:
The inhalatory LC50, 4h value of Diformal in Wistar rats was established to be within the range of
0.05 – 0.5 mg/L.
Executive summary:

Assessment of acute inhalation toxicity withDIFORMALin the rat (acute toxic class method)

 

 

The study was carried out based on the guideline described in Organisation for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects. No.436, "Acute Inhalation Toxicity-Acute Toxic Class Method", September 2009.

 

 

DIFORMALwas aerosolized at concentrations of 0.55 and 0.055 mg/L. Two groups of three male and three female Wistar rats each were exposed by inhalation to one of these concentrations for a single period of 4 hours. Animals were subjected to daily observations and weekly determination of body weight.Macroscopic examination was performed on the day of death or at terminal sacrifice (day 15).

 

 

The mean actual concentrations were 0.55 ± 0.07 mg/L and 0.058 ± 0.011 mg/L. The nominal concentrations were 1.895 and 0.99 mg/L and the generation efficiencies (ratio of actual and nominal concentration) were 29 and 6% respectively.

 

During the exposure to 0.55 mg/L, the Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. The MMAD was 3.5mm and 4.1mm respectively and the gsd was 1.6 and 1.5 respectively. The MMAD of the second measurement was just abovethe recommended range of 1 – 4µm. Since one MMAD value was within the range and the other close to the upper limit and since the gsd were appropriate (i.e. between 1.5 and 3), it can be assumed that deposition of particles in the lower respiratory tract had occurred.For the 0.055 mg/L, an attempt was made to determine the MMAD and gsd but due to the very small amount of aerosol present (approximately 0.008 mg/L) the measurement did not show a normal distribution and therefore no MMAD and gsd were calculated.

 

All animals exposed to 0.55 mg/L were found dead between Days 1 and 2 post-exposure. No mortality occurred at 0.055mg/L.

 

During exposure, no clinical signs were noted. After exposure, lethargy, hunched posture, ventral-lateral recumbence, slow breathing and/or piloerection were noted among the animals exposed to 0.55 mg/L. No clinical signs were noted in the animals exposed to 0.055 mg/L.

 

Overall body weight gain in surviving males and females were within the range expected for rats of this strain and age used in this type of study

 

No treatment abnormalities were found at macroscopic post mortem examination of the animals.

 

 

The inhalatory LC50, 4hvalue of Diformal in Wistar rats was established to be within the range of

0.05 – 0.5 mg/L.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Quality of whole database:
One K1 study is available.

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 August 2013 - 21 August 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD guidelines and GLP.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: CRL:(WI)
Sex:
male/female
Details on test animals and environmental conditions:
Species and strain: CRL:(WI) Wistar rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
Hygienic level at arrival: SPF
Hygienic level during
the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies
Number of animals: 5 animals/sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at study start: Young adult rats
Body weight range
at dosing: Between 217 g and 256 g
Acclimatization time: 6 days

Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room-Box: 242/7
Housing: Individual caging
Cage type: Type II. polypropylene/polycarbonate
Bedding: Laboratory bedding:
Lignocel Hygienic Animal Bedding produced by J. Rettenmaier & Söhne GmbH+Co.KG (Holzmühle 1, 73494 Rosenberg, Germany);
A copy of the relevant Certificate of Analysis is maintained in CiToxLAB Hungary Ltd.'s archive.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20.5 - 29.7 °C
Relative humidity: 30 - 70 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
The temperature and relative humidity was 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 not considered to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
The batch of feed employed in the study was as follows:
 175 8935, expiry date: August 2013
The supplier provided an analytical certificate for the batch used. Copy of the certificate will be archived with the raw data.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment. The test item was applied as a single dose as supplied to the shaved skin and remained in contact with the skin for the 24- hour exposure period. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.
At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
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 day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the 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 weights were recorded on Day 0 (before test item administration) and on Days 7 and 14. Moreover, the body weight of the found dead animal was recorded at necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: None
Statistics:
None
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Remarks on result:
other: 1 female died on day 1
Mortality:
Administration of the test item at a dose level of 2000 mg/kg body weight caused mortality in 1/10 animal on day 1.
Clinical signs:
No clinical signs were observed after treatment with the test item or during the 14-day observation period. There were no observed local dermal signs after treatment with the test item or during the 14 day observation period at a dose level of 2000 mg/kg bw.
Body weight:
The body weight and body weight gain of treated animals did not show any test item-related effect.
Gross pathology:
Dark/red discoloration/foci of the non-collapsed lungs, foamy red material in the trachea and thymus and red macule at the site of application were found in this female rat at necropsy.
There was no evidence of the any gross observations in surviving animals at a dose level of 2000 mg/kg bw and terminated on Day 14.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal median lethal dose (LD50) of the test item PREPOLYMER D was found to be higher than 2000 mg/kg bw in male and female CRL:(WI) rats.
Executive summary:

An acute dermal toxicity study was performed with the test item in CRL:(WI) rats, in compliance with OECD Guideline No.: 402.

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied as a single dermal 24-hour exposure followed by a 14-day observation period.

Clinical observations were performed on all animals at 1 and 5 hours after dosing and on the surviving animals daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14 and of the animal found dead. Gross macroscopic examination was performed on the surviving animals at the end of the 2-week observation period (Day 14). Moreover, the animal found dead was examined macroscopically and body weight was recorded at necropsy.

The results of the study were summarized as follows:

Mortality

Administration of the test item at a dose level of 2000 mg/kg body weight caused mortality in 1/10 animal.

Systemic clinical signs

No clinical signs were observed after treatment with the test item or during the 14-day observation period.

Local dermal signs

There were no observed local dermal signs after treatment with the test item or during the 14 day observation period at a dose level of 2000 mg/kg bw.

Body weight

The body weight and body weight gain of treated animals did not show any test item-related effect.

Necropsy

Dark/red discoloration/foci of the non-collapsed lungs, foamy red material in the trachea and thymus and red macule at the site of application were found in this female rat at necropsy.

There was no evidence of the any gross observations in surviving animals at a dose level of 2000 mg/kg bw and terminated on Day 14.

Conclusions

The acute dermal median lethal dose (LD50) of the test item was found to be higher than 2000 mg/kg bw in male and female CRL:(WI) rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Quality of whole database:
One K1 study is available.

Additional information

Justification for selection of acute toxicity – oral endpoint

K1: The study was performed according to OECD guidelines and GLP.

Justification for selection of acute toxicity – inhalation endpoint

K1: The study was performed according to OECD guidelines and GLP.

Justification for selection of acute toxicity – dermal endpoint

K1: The study was performed according to OECD guidelines and GLP.

Justification for classification or non-classification

The oral LD50 is between 50 and 300 mg/kg bw. Therefore the substance is classified category 3, Toxic if swallowed.

The dermal LD50 is >2000 mg/kg bw. Therefore the substance is not classified for acute dermal toxicity.

Based on the very low vapour pressure of the substance and a pattern of use that does not result in the formation of aerosoles an acute inhalation study was not performed. However, an acute inhalation study is available for the major constituent of the substance, 40% is bis(2-chloroethoxy)methane CAS 111 -91 -1 + homologues. The inhalatory LC50, 4h value for this substance is within the range of 0.05 – 0.5 mg/L leading to category 2 classification, the same C&L will be applied to the multiconstituent substance.