Reaction mass of tetrasodium 7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(2-methyl-4-sulphonatophenyl)azo]naphthalene-2-sulphonate] and tetrasodium 4-[[1-hydroxy-6-[[[[5-hydroxy-6-[(2-methyl-4-sulphonatophenyl)azo]-7-sulphonato-2-naphthyl]amino]carbonyl]amino]-3-sulphonato-2-naphthyl]azo]benzoate and tetrasodium 4,4'-[carbonylbis[imino(1-hydroxy-3-sulphonatonaphthalene-6,2-diyl)azo]]dibenzoate

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nosan No. 8147

GLP compliance:

yes (incl. QA statement)

Remarks:

Bioassay Labor für biologische Analytik GmbH, 69120 Heidelberg, Germany

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Tetrasodium 4-[[1-hydroxy-6-[[[[5-hydroxy-6-[(2-methyl-4-sulphonatophenyl)azo]-7-sulphonato-2-naphthyl]amino]carbonyl]amino]-3-sulphonato-2-naphthyl]azo]benzoate, Tetrasodium 4,4'-[carbonylbis[imino(1-hydroxy-3-sulphonatonaphthalene-6,2-diyl)azo]]dibenzoate and Tetrasodium 7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(2-methyl-4-sulphonatophenyl)azo]naphthalene-2-sulphonate]

IUPAC Name:

Reaction mass of Tetrasodium 4-[[1-hydroxy-6-[[[[5-hydroxy-6-[(2-methyl-4-sulphonatophenyl)azo]-7-sulphonato-2-naphthyl]amino]carbonyl]amino]-3-sulphonato-2-naphthyl]azo]benzoate, Tetrasodium 4,4'-[carbonylbis[imino(1-hydroxy-3-sulphonatonaphthalene-6,2-diyl)azo]]dibenzoate and Tetrasodium 7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(2-methyl-4-sulphonatophenyl)azo]naphthalene-2-sulphonate]

Details on test material:

- Name of test material (as cited in study report): Direct Orange 102 similar
- Physical state: Solid / orange to red
- Storage condition of test material: Room temperature, dry storage, no direct sunlight
- Content: About 90 g/100 g
- Lot/batch No.: #0011852462
- Test item No.: 14/0416-1
- Expiry date: May 26, 2020

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Age at study initiation: male animals approx. 8 weeks, female animals approx. 12 weeks
- Mean weight at study initiation (mean weight ± SD): males: 227.6 ± 2.88 and females 206.8 ± 2.17 g
- Housing: Single housing, Makrolon cage, type III with bedding (H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)) and enrichment (Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria)
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70
- Air changes (per hr): Approx. 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5% solution in deionized water

Details on dermal exposure:

TEST SITE
- Area of exposure: Single application to the clipped epidermis (dorsal and dorsolateral parts of the trunk).
- % coverage: approx. 10 (about 40 cm²)
- Type of wrap if used: semi-occlusive dressing.

REMOVAL OF TEST SUBSTANCE
- Washing: rinsing of the application site with warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 5.71 mL/kg bw
- Concentration: 35g/100 mL
- Form of application: suspension
- Preparation test item: The test item preparation was produced for the test group shortly before application by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer. Additionally the homogeneity of the test item preparation during application was ensured by stirring with a magnetic stirrer.

VEHICLE
A good homogeneity in water could not be guaranteed, because the test item preparation was a suspension. Therefore a 0.5% solution of CMC in deionized water was applied.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before application (day 0), weekly thereafter and on the last day of observation.
- Clinical observations: Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter.
- Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), several times and on the last day of observation
- Mortality: A check for any dead or moribund animals was made at least once each workday.
- Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: No systemic clinical signs were observed.

Gross pathology:

No macroscopic pathologic abnormalities were noted in all animals (5 males and 5 females) examined on the last day of observation.

Other findings:

Local effects:
- Males: Due to the reddish discoloration of the application area, no erythema was assessable on study day 1 only. Moderate erythema (grade 3) was seen in four animals from day 2 until day 3, followed by well-defined erythema from day 6 until day 9 and very slight erythema (grade 1) on day 10. The fifth animal showed also moderate erythema from day 2 until day 3, but followed by very slight erythema on day 6, 7 and 10. Four animals showed severe edema (grade 4) on day 1, followed by moderate edema (grade 3) from day 2 until day 3 and very slight edema (grade 1) on day 6 and 7. The fifth male animal showed moderate edema from day 1 until day 3, followed by slight edema (grade 2) on day 6 and 7 and very slight edema on day 8 and 9. Red test item residues and a reddish discoloration of the application area could be noted from day 2 until day 9 in all animals.
- Females: Due to the reddish discoloration of the application area, no erythema was assessable on day 1 only. Moderate erythema (grade 3) was seen in four animals from day 2 until day 3, followed by well-defined erythema (grade 2) from day 6 until day 9 or day 10. One of these animals showed very slight erythema (grade 1) on day 10. In the fifth animal showed well-defined erythema (grade 2) was observed from day 2 until day 10. Four animals showed moderate edema (grade 3) from day 1 until day 2, ensued by slight edema (grade 2) on day 3. On day six very slight edema (grade 1) could be noted in three of these animals, while the fourth animal still showed slight edema on day 6. In the fifth animal moderate edema was observed on day 1, followed by slight edema from day 2 until day 3 and very slight edema on day 6. Red test item residues and a reddish discoloration of the application area could be noted from day 2 until day 10 in all animals.