REAKTIV-ORANGE DYPR 934

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-01-26 to 1999-02-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted in compliance with EEC-Guideline B1 and OECD Guidelines for testing chemicals 401 according to the good laboratory practice regulations annex of paragraph 19a, section 1 of the chemical law of July, 25, 1994.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

Standard acute method

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Details on test material:

-Name of test material (as cited in study report): Reaktiv-Orange DYPR934

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST SYSTEM
Species Sprague Dawley rat
Strain HSD: Sprague Dawley SD
Origin HARLAN WINKELMANN
(SPF breeding colony)
Body weight at start of the experiment
Male animals Mean = 206g (=100%)
S =± 6.1 g
Min= 198 g (-3.9%)
Max = 212 g (+2.9%)
n = 5
Female animals Mean = 179 g (=100%)
S =± 5.4 g
Min= 172 g (-3.9%)
Max = 184 g (+2.8%)
n = 5
Age at the start of the study: 6-10 weeks
Randomization Randomization schemes: 98.0941, 98.0704
Animal maintenance: in fully air-conditioned rooms in macrolon cages (type 4) on soft wood granulate in groups of 5 animals
Room temperature: 22±3°C
Relative humidity: 50± 20%
Lighting time: 12 hours daily
Acclimatization: at least seven days
Food: ssniff® R/M-H (V 1534), ad libitum
Withdrawal of food: from about 16 hours before to 3-4 hours after treatment
Water: tap water in plastic bottles, ad libitum
Animal identification: fur marking with KmnO4

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Deionized water

Details on oral exposure:

VEHICLE
- Concentration in vehicle and amount of vehicle: The animals received the compound as a 20 % solution in deionized water, the administration volume being 10 mL/kg body weight
-Oral gavage to fasted animals:
2000 mg/kg bodyweight (base on a dose range finding study)

Doses:

2000 mg/kg solution in deionized water (10 mL/kg) bw.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

Duration of observation period following administration: 14 days (The observation period following treatment lasted for 14 days)
Frequency of observations and weighing: Symptoms recorded twice every day (in the morning and in the afternoon) on weekends and public holidays, only once. During this time the animals were weighed weekly. At the end of the observation period the animals

Necropsy of survivors performed: yes
Clinicals observations: the following clinical signs were observed after the administration of Reaktiv-Orange DYPR 934:
Squatting posture, stilted gate, irregular respiration, diarrhea and orange discolored feces.
Body weight: development of body weight was not impaired
Lethality: no death occurred during the whole study

Results and discussion

Preliminary study:

No lethality occurred after application of 2000 mg/kg body weight:
Besides unspecific symptoms:
Impairments of mobility and respiration. Additionally diarrhea and orange discolored feces were observed.
Clinical signs: squatting posture, stilted gait, irregular respiration, diarrhea and orange discolored feces.
At day two of the study the irritations were disappeared.

Development of body weight was not impaired
Autopsy findings: no macroscopically visible changes

No symptoms were observed after administration of 500 mg/kg body weight

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0

Clinical signs:

other: No deaths occurred during the whole study. The following clinical signs were observed after the administration of the test substance: squatting posture, stilted gait, irregular respiration, diarrhea and orange discolored feces. At day two of the study t

Gross pathology:

INDIVIDUAL GROSS PATHOLOGY OBSERVATIONS
Group I: Dose: 2000 mg/kg body weight Sex: female
Animal number Mode of Death Death
Day (Week) Observation(s)
6 SCHEDULED KILL: FINAL 15 (3) No visible Lesions
7 SCHEDULED KILL: FINAL 15 (3) No visible Lesions
8 SCHEDULED KILL: FINAL 15 (3) No visible Lesions
9 SCHEDULED KILL: FINAL 15 (3) No visible Lesions
10 SCHEDULED KILL: FINAL 15 (3) No visible Lesions



The animals killed at the end of the observation period showed no macroscopically visible changes.

Other findings:

- Other observations: No deaths occurred during the whole study.

Any other information on results incl. tables

Dose Range finding study

Test substance: Reaktiv-Orange DYPR 934

Vehicle: dionized water

Concentration: 20%

Dose Mg/kg Body weight	Sex (m/f)	Date of administration	Body weight (g) Days after administration 0      7            14	Lethality     Time                      bw P.a                            [g]
2000	m f	1999-01-26	214   268      295	-	-
		1999-01-26	170   220      220	-	-
1000	m f	1999-01-26	198    243     263	-	-
1000		1999-01-26	158     195    203	-	-
500	m f	1999-01-26	210     267    292	-	-
500		1999-01-26	172     202    217	-	-

Comprehensive description of clinicals signs:

No deaths occurred during the whole study.

The following clinical signs were observed after the administration of the test substance in the 1000 and 2000 mg/kg bw. group: squatting posture, stilted gait, diarrhea and orange discolored feces. At day two of the study the irritations were disappeared. Development of body weight was not impaired.

No symptoms was observed after administration of 500 mg/kg body weight.

Comprehensive description of the macroscopic findings:

The animals killed at the end of the observation showed no macroscopically visible changes.

No lethality occurred after application of 2000 mg/kg body weight.

 

Table I: Bodyweights-Individual Bodyweights Study: 99. 0012-Acute Oral Toxicity with Reaktiv-Orange DYPR 934 in Rats
Table 1: Bodyweights: Individual Bodyweights
Day numbers relative to Start Date
Group Sex		Animal Number	1		6		15
1m		1	212		265*		287
		2	206		261*		269
		3	211		265*		303
		4	201		260*		287
		5	198		258*		296
		Mean	205.6		261.8		292.4
		S.D.	6.1		3.1		7.0
		N	5		5		5
1 f		6	184		208		216
		7	181		207		215
		8	172		197		207
		9	183		210		216
		10	174		203		210
		Mean	178.6		205.0		212.8
		S.D.	5.4		5.1		4.1
		N	5		5		5
*= result to left has an associated comment or marker Nominal Dose: Group 1-2000 mg/kg bodyweight
Comments and Markers
Measurement	Group Sex			Animal number		Day Number		Type		Marker Comment
Bodyweights	1 m			1		8		Out of range		<
				2		8		Out of range		<
				3		8		Out of range		<
				4		8		Out of range		<
				5		8		Out of range		<
Marker= implies value excluded from means
Day numbers relative to Start Date
Table 2: Bodyweights Gains- Individual Bodyweight Gains
	Group Sex			Animal Number		From   To		1   8		8   15
	1m			1				53*		22*
				2				55*		28*
				3				54*		38*
				4				59*		27*
				5				50*		38*
						Mean		56.2		30.6
						S.D.		3.1		7.1
						N		5		5
*= result to left has an associated comment or marker                                          Nominal Dose: Group 1-2000 mg/kg bodyweight
	Group Sex			Animal Number		From   To		1   8		8   15
	1f			6				24		8
				7				26		8
				8				25		10
				9				27		6
				10				29		7
						Mean		26.2		7.8
						S.D.		1.9		1.5
						N		5		5

 

 

Clinical Observations- Table 3: Clinical signs by Animal
Day numbers relative to Start Date
Group Sex	Animal Number	Clinical Sign	I 0-10 min Post dos	I 10-30 min Post dos	I 30-60 min Post dos	1-2 hrs Post dos	I 2-4 hrs Post dos	I 4-8 hrs Pos dos
1 m	1	No Abnormalities Detected	X	X	-	-	-	-
		Gait stilted	-	-	X	X	X	X
		Squatting posture	-	-	X	X	X	X
		Diarrhea	-	-	-	-	X	X
		Feces discolored	-	-	-	o	o	o
		Irregular respiration	-	-	-	-	-	-
	2	No Abnormalities Detected	X	X	X	-	-	-
		Gait stilted	-	-	-	X	X	-
		Squatting posture	-	-	-	X	X	-
		Feces discolored	-	-	-	-	o	o
		Irregular respiration	-	-	-	-	X	-
	3	No Abnormalities Detected	X	X	-	-	-	-
		Gait stilted	-	-	X	X	X	X
		Squatting posture	-	-	X	X	X	X
		Feces discolored	-	-	-	-	o	o
		Irregular respiration	-	-	-	-	X	X
	4	No Abnormalities Detected	X	X	-	-	-	-
		Gait stilted	-	-	X	X	X	-
		Squatting posture	-	-	X	X	X	-
		Feces discolored	-	-	-	-	0	0
5		No Abnormalities Detected	X	X	-	-	-	-
		Gait stilted	-	-	X	X	X	X
		Squatting posture	-	-	X	X	X	X
		Feces discolored	-	-	-	o	o	0
			X
		Diarrhea	X	X	X	X	X	X
			X	X	X	X	X	X
		Irregular respiration	X	X	X	X	X	X
1f	6	No Abnormalities Detected	X	X	-	-	-	-
		Gait stilted	-	-	X	X	X	X
		Squatting posture	-	-	X	X	X	X
		Diarrhea	-	-	-	X	X	X
		Feces discolored	-	-	-	o	o	o
		Irregular respiration	-	-	-	-	X	X
	7	No Abnormalities Detected	X	-	-		-	-
		Gait stilted	-	X	X	X	X	X
		Squatting posture	-	X	X	X	X	X
		Diarrhea	-	-	X	X	X	X
		Feces discolored	-	-	o	o	o	o
	8	No Abnormalities Detected	X	-	-	-	-	-
		Gait stilted	-	X	X	X	X	X
		Squatting posture	-	X	X	X	X	X
		Diarrhea	-	-	X	X	X	X
		Feces discolored	-	-	o	o	o	o
		Irregular respiration	-	-	-	-	X	X
	9	No Abnormalities Detected	X	X	-	-	-	-
		Gait stilted	-	-	X	X	X	X
		Squatting posture	-	-	X	X	X	X
		Diarrhea	-	-	-	X	X	X
		Feces discolored	-	-	-	o	o	o
	10	No Abnormalities Detected	X	-	-	-	-	-
		Gait stilted	-	X	X	X	X	X
		Squatting posture	-	X	X	X	X	X
		Diarrhea	-	-	X	X	X	X
		Feces discolored	-	-	o	o	o	o
		Irregular respiration	-	-	-	-	X	-
X= Present O= orange Nominal Dose: Group I -2000mg/ kg bodyweight  -result to left has an associated comment or marker

 

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on results obtained in this study the median lethal dose value (LD50) of Reaktiv Orange DYPR 934 for the male and female rat is greater than 2000 mg/kg body weight.

Executive summary:

In an acute toxicity oral study (conducted in accordance with the OECD Guideline), Reaktiv-Orange DYPR 934 was administrated by gavage to 5 male and 5 female of Sprague Dawley rats.

The observation period following treatment lasted for 14 days. Symptoms were recorded twice every day (in the morning and in the afternoon) on weekends and public holidays, only once. During this time the animals were weighed weekly. At the end of the observation period the animals were killed by carbone dioxide asphyxiation, dissected and examined for macroscopically visible changes.

The acute oral toxicity testing of Reaktiv-Orange DYPR 934 in the rat yielded a median lethal dose (LD50) above 2000 mg/kg body weight in both male of female animals. No lethality occurred after application of 2000 mg/kg body weight. Besides unspecific symptoms the animals showed impairments of mobility and respiration. Additionally the diarrhea and orange discolored faces were observed. At day two of the study the irritations were disappeared. Development of body weight was not impaired. The animals killed at the end of the observation period showed no macroscopically visible changes.

Results Synopsis and test report:

Species	Strain	Origin	Sex of animals dosed	Tabulation of response data by dose level	LD50 Value	Findings
Sprague Dawley rats	HSD: Sprague Dawley SD	Harlan Winkelmann Gartenstr. 27, 33178 Borchen (SPF breeding colony)	Male and female	No animals died during the test.	>2000 mg/kg body weight	The animals killed at the end of the observation period showed no macroscopically visible changes.