HPSA

Endpoint summary

Description of key information

Acute oral toxicity (1996), GLP, ACUTE ORAL TOXICITY (LIMIT TEST) IN THE RAT (GERMAN WATER HAZARDS CLASSIFICATION SCHEME), gavage, 2000 mg/kg, LD50 >2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 April - 16 May 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material as a solution in distilled water at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed and subjected to gross pathological examination.

GLP compliance:

yes (incl. QA statement)

Remarks:

(The Department of Health of the Government of the United Kingdom)

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd. , Margate , Kent , UK
- Age at study initiation: 5 - 8 weeks
- Weight at study initiation: 157-173g (males) and 133-148g (females)
- Fasting period before study: overnight fast immediately before dosing and for approximately two hours after dosing
- Housing: animals were housed in groups of up to five by sex in solid-floor polypropylene cages furnished with woodflakes
- Diet : Rat and Mouse Expanded Diet No.1 (Special Diets Services Limited , Witham , Essex , UK) , ad libitum
- Water : mains drinking water , ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 43-62
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg bw

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Range-finding Study
A range-finding study was performed to establish a dosing regime with undiluted test material at a dose level of 2000 mg/kg and formulations of 2000 mg/kg and 200 mg/kg in distilled water. The animals were observed for deaths or overt signs of toxicity 1/2, 1, 2 and 4 hours after dosing and subsequently once daily for five days. Individual bodyweights were recorded on the day of dosing to allow calculation of individual treatment volumes. No necropsies were performed.
Main study
Based on the results of the range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material as a solution in distilled water at a dose level of 2000 mg/kg bodyweight. The animals were observed for deaths or overt signs of toxicity 1/2, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Individual bodyweights were recorded prior to dosing on Day 0 and on Days 7 and 14 or at death. At the end of the study the surviving animals were killed by cervical dislocation. All animals were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Statistics:

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test material was made.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Range-finding study :
The two animals treated with the undiluted test material at a dose level of 2000 mg/kg were found dead 30 minutes after dosing. When the test material was administered as a formulation there were no deaths.
Main study :
One female was found dead 30 minutes after dosing.

Clinical signs:

other: Range-finding study : Clinical signs of toxicity noted in the female treated with 2000 mg/kg were ataxia, hunched posture and decreased respiratory rate. There were no signs of toxicity noted in the male treated with 2000 mg/kg or the animals treated with

Gross pathology:

Abnormalities noted at necropsy of the female that died during the study were abnormally red lungs, dark liver, dark kidneys, severe haemorrhage of the gastric mucosa and severe haemorrhage of the small and large intestines. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

TABLE 1 :

INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DATA IN THE RANGE-FINDING STUDY

Dose Level mg/kg	Animal No. and sex	Effects noted after dosing (Hours)				Effects noted during period after dosing (Days)
		½	1	2	4	1	2	3	4	5
2000 *	1-0 Male	X
	2-0 Female	X
2000	3-0 Male	0	0	0	0	0	0	0	0	0
	5-0 Female	0	0	0	0	HARd	0	0	0	0
200	3-1 Male	0	0	0	0	0	0	0	0	0
	4-1 Female	0	0	0	0	0	0	0	0	0

* = test material administered undiluted as supplied

A = ataxia

H = hunched posture

Rd = decreased respiratory rate

X = animal dead

0 = no signs of systemic toxicity

TABLE 2 :

INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DATA IN THE MAIN STUDY

Dose Level mg/kg	Animal No. and sex	Effects noted after dosing (Hours)				Effects noted during period after dosing (Days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	6-0 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	6-1 Male	0	0	0	0	0	0	0	HRdRIRg RnDhEm DaWt	HRdRI RgRnDh EmDaWt	HRdRI RnEmDa DhWt	HRdRI RnEm DaDh	EmH RdRI	HRdRI Em	HRdRI Em	EmRIH Rd	HRdRI	H	0
	6-2 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	6-3 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	6-4 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	7-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	7-1 Female	X
	7-2 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	7-3 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	7-4 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

H = hunched posture

Rn = noisy respiration

Wt = tiptope gait

Rd = decreased respiratory rate

Da = distended abdomen

X = animal dead

RI = laboured respiration

Em = emiciation

0 = no signs of systemic toxicity

Rg = gasping respiration

Dh = dehydration

TABLE 3 :

INDIVIDUAL BODYWEIGHTS AND WEEKLY CHANGES IN THE MAIN STUDY

Dose Level mg/kg		Animal Number and Sex		Bodyweight (g) at Day					Bodyweight (g) at Death	Bodyweight Gain (g) During Week
				0	7		14			1		2
2000		6-0 Male		161	181		249			20		68
2000		6-1 Male		165	131		201			-34		70
2000		6-2 Male		173	203		231			30		28
2000		6-3 Male		157	193		232			36		39
2000		6-4 Male		159	199		251			40		52
2000		7-0 Female		148	191		213			43		22
2000		7-1 Female		133	-		-		133	-		-
2000		7-2 Female		141	169		199			28		30
2000		7-3 Female		146	177		212			31		35
2000		7-4 Female		140	190		209			50		19

- = animal dead

TABLE 4 :

INDIVIDUAL NECROPSY FINDINGS IN THE MAIN STUDY

Dose Level mg/kg		Animal Number and Sex		Time of Death	Macroscopic Observations
2000		6-0 Male		Killed Day 14	No abnormalities detected
		6-1 Male		Killed Day 14	No abnormalities detected
		6-2 Male		Killed Day 14	No abnormalities detected
		6-3 Male		Killed Day 14	No abnormalities detected
		6-4 Male		Killed Day 14	No abnormalities detected
		7-0 Female		Killed Day 14	No abnormalities detected
		7-1 Female		Found dead Day 0	Abnormally red lungs, dark liver, dark kidneys, severe haemorrhage of the gastric mucosa, severe haemorrhage of the small and large intestines
		7-2 Female		Killed Day 14	No abnormalities detected
		7-3 Female		Killed Day 14	No abnormalities detected
		7-4 Female		Killed Day 14	No abnormalities detected

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

A study in 5 male and 5 female rats revealed that acute oral LD50 for the test material was > 2000 mg/kg bw. The study was performed according to the general guidelines for acute oral toxicity under GLP conditions.

Executive summary:

A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley CD strain rat.

The results were used as a basis for classification under the German Water Hazards Classification Scheme. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material as a solution in distilled water at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed and subjected to gross pathological examination. One female was found dead 30 minutes after dosing. Clinical signs of toxicity noted in one male during Days 4 to 13 included distended abdomen, dehydration, emaciation, hunched posture, decreased respiratory rate, gasping, laboured and noisy respiration and tiptoe gait. Surviving animals showed an expected gain in bodyweight during the study except for one male which showed bodyweight loss during the first week and expected gain during the second week of the study. Abnormalities noted at necropsy of the female that died during the study were abnormally red lungs, dark liver, dark kidneys, severe haemorrhage of the gastric mucosa and severe haemorrhage of the small and large intestines. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley CD strain rat was found to be greater than 2000 mg/kg bodyweight. The test material was assigned the numerical value 1 according to the German Water Hazards Classification Scheme.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity:

A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley CD strain rat.

The results were used as a basis for classification under the German Water Hazards Classification Scheme. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material as a solution in distilled water at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed and subjected to gross pathological examination. One female was found dead 30 minutes after dosing. Clinical signs of toxicity noted in one male during Days 4 to 13 included distended abdomen, dehydration, emaciation, hunched posture, decreased respiratory rate, gasping, laboured and noisy respiration and tiptoe gait. Surviving animals showed an expected gain in bodyweight during the study except for one male which showed bodyweight loss during the first week and expected gain during the second week of the study. Abnormalities noted at necropsy of the female that died during the study were abnormally red lungs, dark liver, dark kidneys, severe haemorrhage of the gastric mucosa and severe haemorrhage of the small and large intestines. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

The acute oral median lethal dose (LD50) of the test material in the Sprague-DawleyCD strain rat was found to be greater than 2000 mg/kg bodyweight. The test material was assigned the numerical value 1 according to the German Water Hazards Classification Scheme.

Justification for selection of acute toxicity – oral endpoint
Only one study available.