Dicyclohexylcarbodiimide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

not specified

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Dicyclohexylcarbodiimide

EC Number:

208-704-1

EC Name:

Dicyclohexylcarbodiimide

Cas Number:

538-75-0

Molecular formula:

C13H22N2

IUPAC Name:

N,N'-dicyclohexylcarbodiimide

Specific details on test material used for the study:

- purity: 99.7%

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Co., Ltd. Japan
- Age at study initiation: 5-6 weeks
- Weight at study initiation: 154-171 (male) - 112-132 (female)
- Fasting period before study: from the day before administration to 3 hours after administration.
- Housing: five animals per cage (same sex)
- Diet: Lab Animals Chow E1 (MF Oriental Yeast Co., Ltd.) ad libitum.
- Water: Tap water filtered through a 5 µm filter, ad libitum. (water quality inspections performed regularly)
- Acclimation period: 6-7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25ºC
- Humidity (%): 40-70%
- Air changes (per hr): 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE: The vehicle and dose selection was based on the results of a preliminary test.
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg

Doses:

500 (only females), 700, 1000, 1400 and 2000 mg/kg.

No. of animals per sex per dose:

5 male/ 5 female per group

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations of clinical signs and mortality were performed 15, 30 minutes, 1, 2, 4, 6, 7, and 8 hours after administration on the day of administration, and once a day thereafter (up to 14 days). Body weights were recorded on days 0, 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

1 110 other: mg/kg

Based on:

act. ingr.

95% CL:

>= 832 - <= 1 480

Effect levels 2

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 110 other: mg/kg

Based on:

act. ingr.

95% CL:

>= 877 - <= 1 404

Mortality:

60% of males and females died at concentrations of 1000 and 1400 mg/kg. At 2000 mg/kg, 60% of males and 100% of females died.

Clinical signs:

other: The following clinical signs were observed in both sexes. These signs were increasing in the first eight hours but the survivors recovered completely: decrease of locomotor activity was seen at 1000 mg/kg and above; hypopnoea was detected at 1400 mg/kg an

Gross pathology:

Stomach lesions such as focal thickening of the stomach mucosa, adhesion of the stomach and other abdmoninal organs or ulcer were observed from 700 mg/kg in males and from 500 mg/kg in females at necropsy of the animals after the observation period.
In the dead animal's autopsy, the following observations were made: in the heart, there was atrial stretching; at pulmonary level, congestion, edema and/or bleeding, and endotracheal mucus retention were observed; at stomach level, hemorrhage, stomach erosion, ulcer, distension; bleeding, congestion and/or dilatation of the small intestine were observed and finally, yellowing whitening of the liver.

Any other information on results incl. tables

Cumulative mortality/No. of test animals

Sex

Dose

Time after administration

Days after administration

Mortality rate

mg/kg

15m

30m

1h

2h

4h

6h

7h

8h

1

2

3

4…

14

(%)

male

700

0/5

0/5

0

1000

0/5

1/5

2/5

3/5

3/5

60

1400

0/5

2/5

3/5

3/5

60

2000

0/5

1/5

3/5

3/5

60

female

500

0/5

0/5

0

700

0/5

0/5

0

1000

0/5

2/5

3/5

3/5

60

1400

0/5

3/5

3/5

60

2000

0/5

3/5

5/5

5/5

100

*m: minutes, h: hour(s), d: days.

Mean body weight

Sex	Dose mg/kg		Days after administration
			0	7	14
male	700	Mean SD Number	162 3.6 5	213 4.2 5	280 10.4 5
	1000	Mean SD Number	163 6.5 5	210 31.8 2	282 26.2 2
	1400	Mean SD Number	163 1.3 5	204 0.7 2	265 9.2 2
	2000	Mean SD Number	163 5.8 5	189 14.8 2	266 15.6 2
female	500	Mean SD Number	120 4.7 5	160 8.6 5	187 14.7 5
	700	Mean SD Number	120 6.6 5	157 10.2 5	181 13.0 5
	1000	Mean SD Number	119 4.3 5	150 5.7 2	170 6.4 2
	1400	Mean SD Number	122 6.9 5	156 19.1 2	188 17.7 2
	2000	Mean SD Number	126 4.7 5	AD

*units: g; AD: all animals were dead.

Clinical signs (male): No. of animals with findings / No. of surviving animals

Dose (mg/kg)	Clinical signs		Time after administration
			15 m	30 m	1h	2h	4h	6h	7h	8h
700	Loose stool					2/5
	Normal		5/5	5/5	5/5	3/5	5/5	5/5	5/5	5/5
1000	Decrease of locomotor activity	+	1/5	1/4	2/4		4/4	4/4	4/4	4/4
	Hypopnoea		1/5	1/4	2/4
	Salivation	+	4/5	3/4
	Loose stool				1/4	1/4	1/4		1/4	1/4
	Normal		1/5	1/4	0/4	3/4	0/4	0/4	0/4	0/4
1400	Decrease of locomotor activity	+		4/5	3/5	1/5	4/5	4/5	3/5	3/5
		++		1/5			1/5	1/5
		+++							1/5	1/5
	Hypopnoea			3/5	1/5		1/5	2/5	2/5	1/5
	Salivation	+	2/5
	Loose stool				2/5	3/5	2/5		1/5	2/5
	Tiptoe gait							1/5
	Crouching								1/5
	Prone position									1/5
	Normal		3/5	0/5	1/5	2/5	0/5	0/5	1/5	1/5
2000	Decrease of locomotor activity	+		3/5	5/5	2/5	3/5	3/5	2/4	¾
		++				1/5	2/5	2/5	1/4
		+++							1/4	1/4
	Hypopnoea			2/5	2/5	2/5	2/5	2/5	2/4	1/4
	Salivation	+	1/5	1/5
	Loose stool				1/5	3/5	3/5		3/4	3/4
	Crouching								1/4	1/4
	Normal		4/5	1/5	0/5	0/5	0/5	0/5	0/4	0/4

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe.

 

Clinical signs (male):No. of animals with findings / No. of surviving animals

Dose (mg/kg)

Clinical signs

Days after administration

1

2

3

4

5

6

7

8

9

10

-14d

Normal

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

- 5/5

1000

Decrease of locomotor activity

+

2/3

1/2

1/2

1/2

1/2

1/2

--

1/3

Hypopnoea

2/3

1/2

1/2

Soiled perineal region

1/3

Normal

0/3

1/2

1/2

1/2

2/2

2/2

1/2

1/2

2/2

2/2

- 2/2

1400

Decrease of locomotor activity

+

2/3

++

1/3

Hypopnoea

1/3

½

Normal

0/3

2/2

½

2/2

2/2

2/2

2/2

2/2

2/2

2/2

- 2/2

2000

Decrease of locomotor activity

+

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

Hypopnoea

1/2

2/2

2/2

Soiled perineal region

2/2

2/2

1/2

1/2

Abdominal distention

2/2

Normal

0/2

0/2

0/2

0/2

0/2

0/2

0/2

0/2

2/2

2/2

- 2/2

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe.

 

Clinical signs (female): No. of animals with findings / No. of surviving animals

Dose (mg/kg)	Clinical signs		Time after administration
			15 m	30 m	1h	2h	4h	6h	7h	8h
500	Normal		5/5	5/5	5/5	3/5	5/5	5/5	5/5	5/5
700	Loose stool					2/5	3/5
	Normal		5/5	5/5	5/5	3/5	2/5	5/5	5/5	5/5
1000	Decrease of locomotor activity	+	2/5	4/5		1/5	3/5	4/5	4/5	3/5
		++	1/5	1/5						1/5
	Hypopnoea					1/5	1/5	1/5	1/5	1/5
	Irrgular respiration		1/5
	Salivation	+	2/5	1/5		1/5
		++		1/5	1/5
	Crouching		1/5
	Normal		2/5	0/5	4/5	4/5	2/5	1/5	1/5	1/5
1400	Decrease of locomotor activity	+			1/5	2/5	2/5	2/5	2/5	2/5
		++	2/5	3/5			1/5	2/5	2/5	2/5
	Hypopnoea			2/5			2/5	2/5	2/5	2/5
	Salivation	+	1/5	3/5
	Crouching		2/5	2/5				1/5	1/5	1/5
	Normal		3/5	1/5	4/5	3/5	2/5	1/5	1/5	1/5
2000	Decrease of locomotor activity	+	2/5	2/5	2/5	1/2	2/2	2/2	1/2	1/2
		++	1/5	2/5		1/2			1/2	1/2
		+++		1/5	2/5
	Hypopnoea		4/5	4/5	3/5	1/2	2/2	2/2	2/2	1/2
	Gasping				1/5
	Salivation	+	4/5	4/5	1/5
	Tiptoe gait									1/2
	Crouching									1/2
	Prone position			1/5
	Loose stool					1/2	1/2			1/2
	Normal		1/5	0/5	0/5	0/2	0/2	0/2	0/2	0/2

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe.

  

Clinical signs (female): No. of animals with findings / No. of surviving animals

Dose (mg/kg)

Clinical signs

Days after administration

1

2

3

4

5

6

7

8

9

10

-14d

500

Normal

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

- 5/5

700

Normal

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

- 5/5

1000

Decrease of locomotor activity

+

2/3

1/2

++

1/3

Hypopnoea

2/3

Normal

0/3

1/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

- 2/2

1400

Decrease of locomotor activity

+

2/2

1/2

1/2

1/2

1/2

1/2

1/2

1/2

Hypopnoea

2/2

1/2

2/2

Tiptoe gait

1/2

Normal

0/2

1/2

0/2

1/2

1/2

1/2

1/2

1/2

2/2

2/2

1/2

2000

-

AD

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe; AD: all animals were dead.

Autopsy findings (male):No. of animals with findings / No. of animals examined

	Organ	findings	Dose (mg/kg)
			700	1000	1400	2000
Dead animals	Heart	Dilatation of atrium	-	1/3	0/3	3/3
	Lung	Congestion/edema	-	1/3	0/3	2/3
	Trachea	Mucus stagnation	-	1/3	0/3	0/3
	Stomach	Focal hemorrhage (glandular area)	-	1/3	1/3	2/3
		Erosion/ulcer (glandular area)	-	2/3	2/3	1/3
		dilatation	-	1/3	2/3	1/3
	Small intestine	Focal hemorrhage	-	1/3	0/3	0/3
		Congestion	-	1/3	0/3	0/3
		Dilatation	-	1/3	0/3	0/3
	Liver	Yellowish change / whitish dot	-	1/3	1/3	0/3
	Urinary bladder	Dark-red urine	-	1/3	0/3	0/3
	Normal		-	0/3	0/3	0/3
Surviving animals	Stomach	Focal thickening of mucosa (non-glandular area)	5/5	2/2	2/2	2/2
		Adhesion (stomach and other abdominal organs)	4/5	2/2	2/2	2/2
	Kidney	Dilatation of the renal pelvis	0/5	1/2	0/2	0/2
	Normal		0/5	0/2	0/2	0/2

 *m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe; AD: all animals were dead.

Autopsy findings (female):No. of animals with findings / No. of animals examined

	Organ	findings		Dose (mg/kg)
			500	700	1000	1400	2000
Dead animals	Heart	Dilatation of atrium	-	-	0/3	1/3	3/5
	Lung	Focal hemorrhage	-	-	0/3	0/3	1/5
	Trachea	Mucus stagnation	-	-	0/3	0/3	1/5
	Stomach	Focal hemorrhage (glandular area)	-	-	2/3	1/3	3/5
		Focal hemorrhage (non- glandular area)	-	-	1/3	0/3	0/5
		Erosion/ulcer (glandular area)	-	-	0/3	3/3	1/5
		dilatation	-	-	1/3	1/3	1/5
	Small intestine	Congestion	-	-	1/3	2/3	0/5
		Dilatation	-	-	0/3	0/3	1/5
	Liver	Yellowish change / whitish dot	-	-	1/3	1/3	1/5
	Normal		-	-	0/3	0/3	0/5
Surviving animals	Stomach	Focal thickening of mucosa (non-glandular area)	5/5	5/5	2/2	2/2	-
		Adhesion (stomach and other abdominal organs)	4/5	4/5	2/2	2/2	-
	Kidney	Dilatation of the renal pelvis	0/5	1/5	0/2	0/2	-
	Normal		0/5	0/5	0/2	0/2	-

 *m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe; AD: all animals were dead.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

EU criteria.

Conclusions:

The LD50 for the test substance in the Acute Oral Toxicity test was found to be 1110 mg/kg bw for both sexes.

Executive summary:

The Acute Oral Toxicity Test was performed according to OECD Guideline 401 (GLP Study). Four concentrations of the test substance were assayed in five males (700, 1000, 1400 and 2000 mg/kg) and five in females (500, 700, 1000, 1400 and 2000 mg/kg) in a single dose toxicity test in Crj:CD (SD) rats; 5 male and 5 female per group. All animals were subject to daily observations and weekly determinations of body weight. Necropsy was performed on the day of their death or after sacrifice, at the end of the observation period. 60% of males and females died at concentrations of 1000 and 1400 mg/kg. At 2000 mg/kg, 60% of males and 100% of females died. Decreased locomotor activity was seen at 1000 mg/kg and above; hypopnoea and loose stool were detected at 1400 mg/kg and above. These signs increased in the first eight hours but survivors recovered completely. The body weight gain was lower in the 2000 mg/kg male group (females died) and in 1400 mg/kg group in both sexes. Autopsies showed stomach lesions at all doses tested. The LD50 for the test item in rats was found to be 1110 mg/kg bw for both sexes.