REAKTIV-ORANGE DYPR 934

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-02-03 to 1999-02-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted in compliance with EEC-Guideline B3 and OECD Guidelines for testing chemicals 402 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 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

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 = 269 g (= 100 %)
S = ± 15.8 g
Min= 241 g (-10.4 %)
Max = 280 g (+4.1 %)
n = 5
Female animals Mean = 208 g (= 100 %)
S = ± 5.5 g
Min = 201 g (-3.4 %)
Max = 213 g (+2.4 %)
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 3) on soft wood granulate one animal per cage.
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
Water: tap water in plastic bottles, ad libitum
Animal identification: cage numbering

Administration / exposure

Type of coverage:

occlusive

Vehicle:

other: Deionized water

Details on dermal exposure:

Before the dermal treatment the fur was mechanically removed from the dorsal skin of the animals over an area of approximately 30 cm2.
The appropriate amount of the test substance was moistened (0. 5 g of Reaktiv-Orange DYPR 934 with 0.5 mL of deionized water.)On two-ply gauze and aluminium foil (6X8 cm) and distributed as uniformly as possible. Together with the foil the test substance was administrated to the shaved and intact dorsal skin. The foil was held in place with an elastic plaster bandage fixed around the animal's body.
At the end of the dermal exposure period of 24 hours the bandage was removed and the treated skin area washed with warm water in order to remove any unabsorbed remnants of the substance.

Duration of exposure:

24 h

Doses:

2000 mg/kg bw.

No. of animals per sex per dose:

5 male and 5 female rats

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During the time the animals were weighed weekly.
- Necropsy of survivors performed: yes
- Other examinations performed:
Body weight development and clinical signs:
With exception of one female, all animals exhibited body weight gain throughout the study. No symptoms were observed after administration of 2000 mg/kg body weight.
The skin of the animals showed orange discolorations.
Autopsy findings:
The animals killed at the end of the observation period showed no macroscopically visible changes

Results and discussion

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 substance related toxical effects.

Gross pathology:

The animals killed at the end of the observation period showed no macroscopically visible changes/ (lesions).

Other findings:

Orange discolored skin

Any other information on results incl. tables

Table I:             RESULTS AND COMMENTS
Lethality	No deaths occurred during the whole study
Body weight development	With exception of one female, all animals exhibited body weight gain throughout the study. No symptoms were observed after administration of 2000 mg/kg body weight.
Clinical signs	No symptoms were observed after administration of 2000 mg/kg body weight. -The skin of the animals showed orange discolorations.
Autopsy findings	The animals killed at the end of the observation period showed no macroscopically visible changes/ (lesions).
Remarks: for individual data on clinical signs, bodyweight development and individual autopsy see tables under the rich text field: “Any other information on results incl. tables”

Dose Range finding study

Test substance: Reaktiv-Orange DYPR 934

Vehicle: dionized water

Preparation of test compound:

To check the suitability of the vehicle 0.5 g of Reaktiv-Orange DYPR 934 was moistened with 0.5 mL deionized water. For individual dosing, both test item and vehicle were mixed in proportion stated above.

Table II: Bodyweights-Individual Bodyweights Study: 99. 0013-Acute Dermal Toxicity with Reaktiv-Orange DYPR 934 in Rats
Bodyweights: Individual Bodyweights
Day numbers relative to Start Date
Group Sex		Animal Number	1		8		15
1m		1	273		298		313
		2	280		296		311
		3	241		245*		263*
		4	277		288		292
		5	272		293		316
		Mean	268.6		284.0		299.0
		S.D.	15.8		22.1		22.2
		N	5		5		5
1 f		6	212		221		220
		7	204		214		222
		8	213		219		224
		9	212		221		233
		10	201		207		216
		Mean	208.4		216.4		223.0
		S.D.	5.5		6.0		6.3
		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			3		8		Out of range		<
						15		Out of range		<
Marker= implies value excluded from means
Day numbers relative to Start Date
Table III: Bodyweights Gains- Individual Bodyweight Gains
	Group Sex			Animal Number		From   To		1   8		8   15
	1m			1				25		15
				2				16		15
				3				4*		18*
				4				11		4
				5				21		23
						Mean		15.4		15.0
						S.D.		8.9		7.0
						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				9		-1
				7				10		8
				8				6		5
				9				9		12
				10				6		9
						Mean		8.0		6.6
						S.D.		1.9		4.9
						N		5		5

 

 Table IV: Individual bodyweight gains

Measurement	Group Sex			Animal number		Day Number		Type		Marker Comment
Bodyweights	1 m			3		8		Out of range		<
						15		Out of range		<

 

 

Table V:                        Clinical observations-Clinical Signs by Animal                                           Day number relative to start
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	X	X	X	X
	2	No abnormalities Detected	X	X	X	X	X	X
	3	No abnormalities Detected	X	X	X	X	X	X
	4	No abnormalities Detected	X	X	X	X	X	X
	5	No abnormalities Detected	X	X	X	X	X	X
	6	No abnormalities Detected	X	X	X	X	X	X
	7	No abnormalities Detected	X	X	X	X	X	X
	8	No abnormalities Detected	X	X	X	X	X	X
	9	No abnormalities Detected	X	X	X	X	X	X
	10	No abnormalities Detected	X	X	X	X	X	X
*= result to left has an associated comment or marker  X=Present

 

Table VI:                  Clinical observations-Clinical Signs by Animal                                          Day number relative to start
Group Sex	Animal Number	Clinical Sign	2	3	4	5	6	7	8	9	10	11	12	13	14	15
1 m	1	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	2	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	3	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	4	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	5	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	6	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	7	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	8	Skin discolored-large area	o	o	o	o	o	o	o	o	o	o	o	o	o	o	o
	9	No Abnormalities Detected	-	-	-	-	-	-	X	X	X	X	X	X	X	X	X
		Skin discolored –large area	o	o	o	o	o	o	-	-	-	-	-	-	-	-	-
	10	Skin discolored –large area	o	o	o	o	o	o	o	o	o	o	o	o	o	o	o

 X= present o= orange

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

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

Executive summary:

In a 24 h acute toxicity dermal study (conducted in accordance with the OECD Guideline as well as the EEC Guideline), the acute dermal toxicity of Reaktiv-Orange DYPR 934 was tested only at a dose level of 2000 mg/kg body weight of 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 dermal 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. After administration of 2000 mg/kg body weight neither death nor symptoms occurred. The skins of eight animals show orange discolorations up to day seven of the study. With exception of one female, all animals exhibited body weight gain throughout the study. The animals killed at the end of the observation period showed no macroscopically visible changes. The dermal LD50 value was considered in conjunction with the observed toxic effects and the necropsy findings. Based on the results obtained in the study the median lethal value (LD50) of Reaktiv-Orange DYPR 934 for the male and female rat is greater than 2000 mg/kg body weight.

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.