Registration Dossier

Administrative data

Endpoint:
acute toxicity: other routes
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
significant methodological deficiencies

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1988

Materials and methods

Test guideline
Qualifier:
no guideline available
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Specific details on test material used for the study:
Obtained by extraction from natural ingredients (citrus peel).
Analytical methods: GLC pattern

Test animals

Species:
mouse
Strain:
ICR
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 5-6 weeks old.
- Fasting period before study: 8h prior to dosing.
- Housing: polycarbonate cages.
- Food (Hwahan pellet feed) and water ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.8 ± 1.9ºC
- Humidity (%): 72,3 ± 7.2%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 9h light /15h dark.

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
propylene glycol
Details on exposure:
Concentration up to 20% (w/v) in propylene glycol.
Doses:
2570, 1977, 1521, 1170, and 900 mg/kg.
No. of animals per sex per dose:
5
Control animals:
yes
Details on study design:
Examinations:
- General clinical observations and body weight, weekly.
- Gross necropsy.
- Hematology: 24 hours after treatment, the animals were anesthetized with urethane. Blood was collected as part of the procedure, centrifuged (5 min at 3000 rpm), and serum separated. Parameters measured: total protein creatinine, GPT, phosphorus, calcium, blood urea nitrogen (BUN), GOT, alkaline phosphatase, and albumin.
- Histopathology.
Statistics:
Statistical analysis method of Litchfield and Wilcoxon was used (Litchfield et al., 1949).

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 650 mg/kg bw
Based on:
test mat.
95% CL:
>= 1 375 - <= 1 980
Remarks on result:
other: intraperitoneal.
Mortality:
The amount of dead animals after the administration of the test item was dose-related: at 2570 mg/kg, all animals died; at 1977 mg/kg, 6/10 animals died; at 1521 mg/kg, 4/10 animals died; at 1170 mg/kg, 2/10 animals died; and at 900 mg/kg, no mortality was observed.
Clinical signs:
No effects.
Body weight:
No effects.
Gross pathology:
No effects.
Other findings:
Hematology: no statistically significant differences were observed.
Histopathology: no statistically significant differences were observed.

Any other information on results incl. tables

Table 1. 50% lethal dose.

 

Dose (mg / kg)

1

2

3

4

5

6

7

Mortality

LD50(mg/kg)

(Confidence limit)

2570

9

1

0

0

0

0

0

10/10

1,650

1977

6

0

0

0

0

0

0

6/10

(1375-1980)

1521

2

2

0

0

0

0

0

4/10

 

1170

2

0

0

0

0

0

0

2/10

 

900

0

0

0

0

0

0

0

0/10

 

(P=0.05 )

 

Table 2. Haematological examination.

Dose

(mg/kg)

Total Protein (g/dl)

Crea­

tinine

(mg/dl)

GOT(IU/Z)

Phosphorus

(mg/dl)

Calcium

(mg/dl)

BUN

(mg/dl)

GPT (IU/l)

alkaline

phosphatase

(IU/Z)

Albumin

(g/dl)

Control

5.27

0.93

74.8

11.6

10.2

30.2

15.6

117.6

2.92

 

± 0.23

± 0.05

± 22.2

± 0.9

± 0.9

± 8.6

± 2.7

± 12.2

± 0.83

50

5.30

0.89

70.6

11.9

1111.0

32.6

18.5

111.4

2.93

 

± 0.20

± 0.10

± 26.2

± 1.0

± 1.0

± 8.2

± 8.2

± 16.2

± 0.42

100

5.34

0.92

77.4

12.2

10.4

30.5

15.5

104.6

2.92

 

± 0. 31

± 0.09

± 25.6

± 1.2

± 0.6

± 5.5

± 7.3

± 20.7

± 0.52

 

Applicant's summary and conclusion

Conclusions:
The LD50 of the test item in mice by ip administration was found to be 1650 mg/kg bw.
Executive summary:

To determine the acute intraperitoneal toxicity of the test item in mice, the test item was administered intraperitoneally at doses of 2570, 1977, 1521, 1170, and 900 mg/kg test item to 5 ICR mice. The LD50 of the test item in mice by ip administration was found to be 1650 mg/kg bw.