Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1987-11-25 - 1987-12-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EPA OPP 81-1 (Acute Oral Toxicity)
Version / remarks:
revised Nov. 1984
Deviations:
no
GLP compliance:
yes
Remarks:
according to US FDA (21 CFR 58) and US EPA (40 CFR 160 and 40 CFR 792)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Appearance: Black powder
Bulk density: 0.21 g/mL
Solubility: Water: approximately 1 ppm; acetone: 0.81 g/100mL (20 °C); chloroform: 2 g/100mL (20 °C)
Storage: At ambient temperature in the dark in closed container
Stability: At least 14 days at 55 °C; poor in organic solvents

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
Young adult rats of the Wistar strain (8 weeks old at study start, SPF-quality, randomly bred) were supplied by Charles River Wiga GmbH, Sulzfeld, FRG. Upon receipt each animal was identified with an ear tag. At least five days prior to dosing (acclimatization period) the animals were individually housed in polycarbonate cages containing purified sawdust (Woody clean supplied by Broekman Institute, Someren, The Netherlands) as bedding material. The animals had free access to tap-water (via automatic-nozzles) and a standard pelleted laboratory animal diet (RMH-B, Hope Farms, Woerden, The Netherlands). Certificates of analysis for both diet and drinking water are retained in the N0T0X archives.

ENVIRONMENTAL CONDITIONS
The animal room was air-conditioned, with the temperature maintained within the range of 20-22°C and the relative humidity within the range of 55-70% during the study. The artificial light cycle was 12 hours light, 12 hours dark.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
methylcellulose
Details on oral exposure:
Animals were fasted overnight prior to dosing until 3 hours after administration of the test substance. The test substance was suspended in 1% aqueous methyl cellulose (Boom B.V.. Meppel, The Netherlands) and administered once only by gavage using a stainless steel stomach cannula attached to a disposable plastic syringe. The dose level was 5000 mg/kg body weight. Each time the dose volume was 15 ml/kg body weight. The day of dosing was designated as day 0.
Doses:
5000 mg/kg body weight
No. of animals per sex per dose:
5 males
5 females
Control animals:
no
Details on study design:
Observations
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on the day of dosing (approximately once every two hours) and once daily thereafter for 14 days
- Necropsy of survivors performed: yes
Statistics:
not perfomed

Results and discussion

Preliminary study:
In order to establish an appropriate dose range three groups of animals, each comprising 1 male and 1 female, were dosed with an oral dose of test substance at 3200, 4200 and 5600 mg/kg body weight. No mortality occurred during the 7-day observation period. Signs of toxicity were body weight loss, lethargy, piloerection, bloody eye encrustation and diarrhoea. Based on the absence of mortality and the low toxicity, observed in the pilot study, the test could be performed as a limit test. One group of animals, comprising 5 males and 5 females, was treated with a single oral dose of test substance at 5000 mg/kg body weight.
Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
One female rat died on day 4 of dosing.
Clinical signs:
Signs of toxicity were body weight loss for the female rat found dead, lethargy, piloerection, dacryorrhoea, diarrhoea and emaciation.
Body weight:
Signs of toxicity were body weight loss for the female rat found dead. All surviving animals showed body weight gain during the study period (Table 2). However, totally 5 animals revealed body weight loss or stagnant body weight during the first week of observation.
Gross pathology:
Macroscopic examination of animals at termination did not reveal any abnormalities that were considered test substance related, with the exception of a slightly enlarged liver of the female rat which died on day 4.
Other findings:
none

Any other information on results incl. tables

Table 1: Daily Incidence of Mortality and Clinical Observations fo Male and Female Rats Combined Following Acute Oral Dosing of test material
Dose level (mg/kg) Signs of reaction Number showing effects during day of dosing (hours:minutes) or day of observation (day) Total mortality
00:15 02:15 03:35 1 2 3 4 5 6 7 8 9 10 11 12 13 14
5000 No abnomalities 10 0 0 1 3 9 4 3 4 5 8 9 9 9 9 9 9
Lethargy 0 10 10 0 0 1 5 1 0 0 0 0 0 0 0 0 0
Emaciation 0 0 0 0 0 0 0 0 2 2 0 0 0 0 0 0 0
Piloerection 0 6 10 1 7 1 0 6 5 3 1 0 0 0 0 0 0
Diarrhoea 0 0 0 9 0 0 0 0 0 0 0 0 0 0 0 0 0
Dacryorrhoea 0 5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Deads 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1/10

Table 2: Indivudual body weights and dose volumes acute oral toxicity study of test material in the rat
Dose Level (mg/kg) Animal number Body Weight (g) on: Body weight Gain (g) Day 0-14 Dose Volume (mL)
Day 0* Day 7 Day 14
5000
(Males)
3306 221 220 304 83 3.3
3308 216 224 291 75 3.2
3310 230 221 301 71 3.5
3312 225 251 313 88 3.4
3314 227 227 303 76 3.4
Mean 224 229 302 79
S.D. 5.4 12.8 7.9 6.8
N 5 5 5 5
5000
(Females)
3113 169 174 218 49 2.5
3115 170 178 222 52 2.6
3117 176 - - - 2.6
3119 174 170 219 45 2.6
3121 185 185 252 67 2.8
Mean 175 177 228 53
S.D. 6.4 6.4 16.3 9.6
N 5 4 4 4
* Day of dosing

Applicant's summary and conclusion

Interpretation of results:
other: EU GHS criteria not met
Conclusions:
LD50 value for both males and females was noted as exceeding 5000 mg/kg.
Executive summary:

The acute oral toxicity of the test item was evaluated in this single-dose study in rats according to EPA OPP 81 -1 (Acute oral toxicity) and in compliance with GLP ( US FDA 21 CFR 58 and US EPA 40 CFR 160 and 40 CFR 792).

One group of Wistar rats, comprising 5 males and 5 females, was treated with a single oral dose of the test material at 5000 mg/kg body weight. One female rat died on day 4 of dosing. Signs of toxicity were body weight loss for the female found dead, lethargy, piloerection, dacryorrhoea, diarrhoea and emaciation. These signs were considered reversible since as of day 9 no more abnormalities were observed during the 14-day observation period. All animals showed body weight gain during the study period. Macroscopic examination of animals at termination did not reveal any abnormalities that were considered test substance related, with the exception of a slightly enlarged liver of the female which died on day 4. Since only one animal died, the LD50 value for both males and females was noted as exceeding 5000 mg/kg body weight.