Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2 weeks
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Not according to OECD guidelines (tested in 1978), but very well documented.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1978
Report Date:
1978

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Test system: New Zealand White albino rabbits; 8 ¿ and 8 ¿, divided in 4 groups according to body weight. Part of the trunk of animals was freed form hair (approx. 10% total body surface).
Dose levels: 0.0, 0.35, 0.7, and 1.4 ml/kg bw; all dose levels were applied in the same volume of 9 ml/kg bw by appropriate diluting with water. Half of the number of animals received the material on the intact skin, the other half on the abraded skin. The treated area was covered with a thin layer of cellose sheet and wrapped in polyethylene foil. After an exposure time of 24 hours the test substance was removed from the skin with water and the animals were wiped dry with towels. Subsequently they were caged individually in a room of constant temperature (18°C) and provided with the standard laboratory diet and tap water ad libitum. After treatment the rabbits were observed for 2 weeks with regard to general appearance and behavior, mortality, local skin reactions, growth, and food and water intake.
At the end of the experimental period examinations were carried out for possible changes in blood composition and macroscopic appearance of several organs. Samples of the heart, liver, kidneys, spleen and of the treated and untreated skin were collected and fixed in a 4% neutral, phosphate-buffered formaldehyde solution for histological examination.
GLP compliance:
not specified
Test type:
other: single dose effect study

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
A sample of the test substance, a colourless solid with a melting point of 27°C, designated 1,4-butanediamine, was received from the principal on 9th November 1977. It was stored in the dark and at room temperature until use.

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
New Zealand White albino rabbits; 8 ¿ and 8 ¿, divided in 4 groups according to body weight. Part of the trunk of animals was freed form hair (approx. 10% total body surface).
After exposure they were caged individually in a room of constant temperature (18°C) and provided with the standard laboratory diet and tap water ad libitum.

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
The test material was melted and put into contact with the shaved skin as aqueous solutions at dose levels of 0.0, 0.35, 0.7, and 1.4 ml/kg bw. All dose levels were applied in the same volume of 9 ml/kg bw by appropriate diluting with water. Half of the number of animals received the material on the intact skin, the other half on the abraded skin. The treated area was covered with a thin layer of cellose sheet and wrapped in polyethylene foil. After an exposure time of 24 hours the test substance was removed from the skin with water and the animals were wiped dry with towels.
Duration of exposure:
24 hours
Doses:
0.0, 0.35 ml/kg bw (307 mg/kgbw), 0.7 (614 mg/kg bw), and 1.4 ml/kg bw (1228 mg/kg bw)
No. of animals per sex per dose:
2 male and 2 female per dose.
Control animals:
yes
Details on study design:
After treatment the rabbits were observed for 2 weeks with regard to general appearance and behavior, mortality, local skin reactions, growth, and food and water intake. At the end of the experimental period examinations were carried out for possible changes in blood composition and macroscopic appearance of several organs. Samples of the heart, liver, kidneys, spleen and of the treated and untreated skin were collected and fixed in a 4% neutral, phosphate-buffered formaldehyde solution for histological examination.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
approximate LD50
Effect level:
> 614 - < 1 228 mg/kg bw
Remarks on result:
other: Value based on lineair extrapolation: 825 mg/kg bw
Mortality:
Mortality amounted to 25% at 0.35 (=307 mg/kg bw)and 0.7 ml/kg bw (=614 mg/kg bw), and to 100% at 1.4 ml/kg bw (=1228 mg/kg bw).
The rabbits of the high dose group were in a poor condition in the course of the exposure period. They died or were killed when moribund during the exposure period or on the first day of the subsequent observation period. In the course of the observation period 2 control rabbits, on rabbit of the 0.35 ml/kg dose group and 1 of the 0.7 ml/kg dose group died, from causes apparently not related to treatment.
Clinical signs:
• 0.0 ml/kg (control group): no abnormalities.
• 0.35 ml/kg (=307 mg/kg bw): slight to severe erythema, slight edema and hemorrhages.
• 0.7 ml/kg (614 mg/kg bw): moderate to severe erythema, moderate to severe edema and extensive hemorrhages.
• 1.4 ml/kg (1228 mg/kg bw): severe erythema, moderate to severe edema, extensive hemorrhages, apathy, sialorrhoea, dyspnea, dark discoloration of irises, paresis and/or paralysis.

Three of the rabbits, the two test rabbits and one control, had suffered from diarrhea.

At the end of the observation period the following dermal effects could still be observed:
• 0.35 ml/kg: distinct scaliness and some necrotic skin areas. Where the necrotic old skin had come off, new skin was visible on which hair growth was disturbed.
• 0.7 ml/kg: distinct scaliness and extensive necrosis. Where parts of the necrotic old skin had come off, new, and sometimes inflamed, skin was visible on which hair growth was disturbed or absent.
There were no distinct differences in reactions either between rabbits treated on the intact or abraded skin, or between male and female rabbits.

Body weight:
Growth figures of the surviving test rabbits, especially of those of the 0.7 ml/kg dose group, were slightly disturbed during the first week of the observation period. Food and water intake of the test group showed some variation but significant differences with the controls did not occur.
Gross pathology:
At autopsy the treated skin of the test animals was found to be distinctly affected by the compound under study.
No other changes that could be ascribed to treatment were observed. Eight of the animals, that died during the experiment, showed signs of diarrhea.One of the control rabbits also showed hemorrhagic erosions in the stomach.
At microscopic examination, treatment-related histopathological changes were found in the treated skin of animals of the low- and intermediate dose group. Animals of the high-dose group did not show any skin lesions, probably because they died within 2 days after the application of the test substance. The skin lesions were mainly characterized by necrosis, acanthosis, infiltration of mainly mononuclear inflammatory cells in the dermis, and fibroblastic repair activity. Microscopy of the liver, kidneys, heart, and spleen did not disclose any abnormalities that could be related to treatment. Gross and microscopic pathology did not provide any information to establish the cause of death of the animals that died during the observation period.
Other findings:
Growth was slightly disturbed at 0.35 and 0.7 ml/kg bw.
Food and water intake and haematology were not affected by 1,4 -butanediamine.
Microscopic examination at autopsy revealed treatment related changes in the skin at 0.35 and 0.7 ml/kg bw.
There were no distinct differences in reactions either between rabbits treated on the intact or abraded skin, or between male and female rabbits.

Any other information on results incl. tables

 dose ml/kg  0.0   0.0   0.0   0.0  0.35  0.35  0.35 0.35   0.7  0.7 0.7  0.7   1.4  1.4  1.4  1.4
 Animal numbers and sex M 7126 M7127  F7135   F7136 M7130   M7131  F7134  F7140  M7123  M7124  F7133  F7141  M7357  M7358  F7361 f7362 
 Died on day:  4    7      12      7        2  2  2
 Gross examination:                                
 1. Diarrhoea      x      x      x        x  x  
 2. Heamorrhagic erosions in the stomach      x                          
 Microscopic examination                                
 Skin                                
 1. Focal Necrosis                                
 a. slight                  x          
 b. moderate            x    x      x  x        
 c. marked

 

                 x            
2. Focal acanthosis                                
a. slight          x  x      x      x        
b. moderate                x      x          
c. marked                    x            
  3.  Slight focal infiltration of mainly mononuclear inflammatory cells in the dermis                    x            
 4. Fibroblastic repair activity in the dermis                    x            
 Liver                                
 1. A few foci of RES cells              x      x            
 2. Slight periportal fibrosis    x                            
 3. Slight centrobular vacuolation of hepatocytes            x                    
 4. Slight periportal infiltration of mainly mononuclear inflammatory cells      x  x    x  x    x    x        
 5. Slight bile-duct proliferation      x                          
 6. A few necrotic hepatocytes  x        x                      
 Kidneys                                
 1. a few dilated tubules      x  x                  x    x  
 2. Slight infiltration of mainly mononuclear inflammatory cells in the cortex          x          x            
3. Slight tubular necrosis   x          x    x  x        
4. A few calcareous deposits in the cortex and/or intercortico medullary layer               x    x    x          
 Heart                                
 1. Sight focal myodegeneration      x                          
 2. Slight focal infiltration of mainly mononuclear inflammatory cells in the myocard  x                  x  x          
                                 
                                 
                                 
                                 

Applicant's summary and conclusion

Interpretation of results:
harmful
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
DSD: Xn Harmful, R22 Harmful in contact with skin
CLP: GHS06 Toxic (cat.3) H311 Toxic in contact with skin
Executive summary:

• 0.0 ml/kg (control group): no abnormalities.

• 0.35 ml/kg (=307 mg/kg): slight to severe erythema, slight edema and hemorrhages.

• 0.7 ml/kg (=614 mg/kg): moderate to severe erythema, moderate to severe edema and extensive hemorrhages.

• 1.4 ml/kg (=1228 mg/kg): severe erythema, moderate to severe edema, extensive hemorrhages, apathy, sialorrhoea, dyspnea, dark discoloration of irises, paresis and/or paralysis.

Mortality amounted to 25% at 0.35 and 0.7 ml/kg bw, and to 100% at 1.4 ml/kg bw.

Growth was slightly disturbed at 0.35 and 0.7 ml/kg bw.

Food and water intake and haematology were not affected by 1,4 -butanediamine.

Microscopic examination at autopsy revealed treatment related changes in the skin at 0.35 and 0.7 ml/kg bw.

There were no distinct differences in reactions either between rabbits treated on the intact or abraded skin, or between male and female rabbits.

DAB: LD50(rabbit, dermal): 0.7-1.4 ml/kg body weight (=614 -1228 mg/kg bw); extrapolated: 825 mg/kg bw

Classification DSD (400< LD50 <2000 mg/kg): Xn Harmful, R22 Harmful in contact with skin.

CLP (200 -1000 mg/kg): GHS06 Toxic (cat.3) H311 Toxic in contact with skin