Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 August 1989 - 26 September 1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in compliance with international guidelines but with deviations: no certificate of analysis of the test substance, animals exposed occasionally to humidity higher than 70%.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
yes
Remarks:
No certificate of analysis; humidity higher than 70%
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
No certificate of analysis; humidity higher than 70%
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): see confidential details
- Physical state: White crystalline powder
- Lot/batch No.: DG6
- Storage condition of test material: at room temperature

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Bantin & Kingman Ltd, Grimston, U.K.
- Age at study initiation: approximatively 5 to 8 weeks old
- Weight at study initiation: males: 120 to 153 g - females: 120 to 150 g
- Fasting period before study: an overnight fast immediately before dosing and for approximately two hours after dosing
- Housing: the animals were housed in groups of up to five by sex in solid-floor polypropylene cages with sawdust bedding
- Diet (e.g. ad libitum): Rat and Mouse Expanded Diet No. 1 (Special Diet Services Limited, Witham, U.K.), ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 54 - 76
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark


IN-LIFE DATES: 24 August 1989 - 26 September 1989

Administration / exposure

Route of administration:
oral: unspecified
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: see table 1
- Amount of vehicle (if gavage): see table 1
Doses:
Range-finding study: 500, 1000, 3000 and 5000 mg/kg
Limit test: 5000 mg/kg
Main study: 2530, 3000, 3557 and 4217 mg/kg
No. of animals per sex per dose:
Range-finding study: 1
Limit test: 5
Main study: 5
Control animals:
no
Details on study design:
- Duration of observation period following administration: range-finding study: 5 days; limit test and main study: 14 days
- Frequency of observations and weighing:
Range-finding study: animals were observed 1 and 4 hours after dosing and subsequently once daily for 5 days. Individual bodyweights were recorded on the day of dosing (day 0). No necropsies were performed.
Limit test and main study: animals were observed 1 and 4 hours after dosing and subsequently once daily for 14 days. Deaths and evidence of over toxicity were recorded at each observation. Individual bodyweights were recorded on the day of dosing (day 0) and on days 7 and 14 or at death. All animals were subjected to gross necropsy examination for any macroscopic abnormalities. No tissues were retained.
Statistics:
The acute oral median lethal dose (LD50) and 95% confidence limits of the test material were calculated using a probit method of Finney D.J. (1971)

Results and discussion

Preliminary study:
No deaths were recorded for all the dose levels in the range-finding experiment.
Effect levelsopen allclose all
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3 354 mg/kg bw
95% CL:
2 834 - 3 970
Sex:
male
Dose descriptor:
LD50
Effect level:
3 264 mg/kg bw
95% CL:
2 648 - 4 024
Sex:
female
Dose descriptor:
LD50
Effect level:
3 494 mg/kg bw
95% CL:
2 654 - 4 601
Mortality:
See table 2
Clinical signs:
Major signs of toxicity noted in all dose groups were hunched posture, pilo-erection, lethargy, ptosis, decreased or gasping respiration and red/brown staining around the snout or mouth. Additional signs of ataxia and/or increased salivation were also noted. Animals treated with 3000, 3557 or 4217 mg/kg also showed distended abdomen and/or emaciation. Animals treated with 3557 mg/kg showed pallor of the extremities with an isolated incident of tiptoe gait.
Surviving animals treated with 2530, 3000 or 5000 mg/kg appeared normal two to six days after treatment. Surviving animals treated with 3557 or 4217 mg/kg appeared normal eight to fourteen days after treatment with one exception of one female treated with 3557 mg/kg which showed signs of toxicity throughout the study period.
Body weight:
Incidents of reduced bodyweight gain or bodyweight loss were noted in all dose groups (see table 3).
Gross pathology:
Abnormalities noted at necropsy of animals that died during the study were haemorrhagic or abnormally red lungs, dark liver or patchy pallor of the liver, pale spleen, pale or dark kidneys, haemorrhage or sloughing of the glandular gastric epithelium and haemorrhage of the small and/or large intestines. One male treated with 3557 mg/kg which died during the study was found cannibalised. The stomach of one male treated with 4217 mg/kg, which died during the study, was adhered to the liver and intestines.
Abnormalities noted at necropsy of animals treated with 2530 mg/kg which were killed at the end of the study were occasional white foci approximately 1 mm x 1 mm covering 75% of the non-glandular gastric epithelium.
No abnormalities were noted at necropsy of animals treated with 3000 mg/kg or greater which were killed at the end of the study.
Other findings:
No data

Any other information on results incl. tables

Table 2: mortality data in the main study

Dose level

mg/kg

Deaths

Male

%

Female

%

Total

%

2530

0/5

0

1/5

20

1/10

10

3000

4/5

80

2/5

40

6/10

60

3557

3/5

60

3/5

60

6/10

60

4217

4/5

80

4/5

80

8/10

80

5000

4/5

80

3/5

60

7/10

70

Table 3: Individual bodyweights in males

Dose level (mg/kg)

Body weight (g) on day :

Increment during week

0

7

14

At death

1

2

2530

141

137

130

142

122

173

150

150

165

154

220

191

190

205

198

 

32

13

20

23

32

47

41

40

40

44

3000

121

120

122

125

124

-

-

-

-

109

-

-

-

-

168

105

98

115

85

 

-

-

-

-

-15

-

-

-

-

59

3557

120

121

122

150

120

199

-

129

156

-

-

-

167

158

-

63

120

 

 

115

-1

-

7

6

-

-

-

38

2

-

4217

120

121

120

121

122

-

130

-

-

140

-

-

-

-

142

90

83

94

104

 

-

9

-

-

18

-

-

-

-

2

5000

150

149

146

153

129

-

-

-

173

-

-

-

-

200

-

103

122

127

 

114

-

-

-

20

-

-

-

-

27

-

Table 4: Individual bodyweights in females

Dose level (mg/kg)

Body weight (g) on day :

Increment during week

0

7

14

At death

1

2

2530

120

120

126

126

150

-

138

137

149

165

-

155

165

179

143

81

 

 

 

 

-

18

11

23

15

-

17

28

30

-22

3000

134

120

139

124

120

123

-

143

129

-

180

-

177

152

-

 

92

 

 

90

-11

-

4

5

-

57

-

34

23

-

3557

126

120

130

138

120

140

-

-

153

142

180

-

-

-

-

 

96

118

131

160

14

-

-

15

22

40

-

-

-

-

4217

145

125

140

123

120

-

-

-

-

131

-

-

-

-

151

142

118

98

80

 

-

-

-

-

11

-

-

-

-

20

5000

133

129

128

126

130

142

-

-

133

-

179

-

-

163

-

 

122

109

 

111

9

-

-

7

-

37

-

-

30

-

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The combined oral LD50 for test material is 3354 mg/kg in rats. As it is higher than 2000 mg/kg bw, it is not classified according to the Annex VI to the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).
Executive summary:

In an acute oral toxicity study performed in accordance with GLP and OECD guideline 401, groups (5/sex) of Sprague Dawley rats were given a single oral dose of test material in arachis oil at 2530, 3000, 3557, 4217 and 5000 mg/kg bw. Animals were then observed for mortality, clinical signs and bodyweights for 14 days and were all macroscopically necropsied after death or sacrifice. A preliminary range-finding test was also conducted on 1 rat/sex/dose at 500, 1000, 3000 and 5000 mg/kg and animals were observed for 5 days. In this previous test, no mortality was recorded.

Mortality occurred in the main study at 3000 mg/kg and higher. Major signs of toxicity noted in all dose groups were hunched posture, pilo-erection, lethargy, ptosis, decreased or gasping respiration and red/brown staining around the snout or mouth. Incidents of reduced bodyweight gain or bodyweight loss were noted in all dose groups. Abnormalities noted at necropsy of animals that died during the study were haemorrhagic or abnormally red lungs, dark liver or patchy pallor of the liver, pale spleen, pale or dark kidneys, haemorrhage or sloughing of the glandular gastric epithelium and haemorrhage of the small and/or large intestines.

The combined oral LD50 was 3354 mg/kg with 95% confidence limits between 2834 and 3970 mg/kg.

The oral LD50 for test material is higher than 2000 mg/kg bw in rats therefore it is not classified according to the Annex VI to the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).