3,10-diamino-6,13-dichloro-2-((6-(((4-(1,1-dimethylethyl)phenyl)sulfonyl)amino)-2-naphthalenyl)sulfonyl)-4,11-triphenodioxazinedisulfonic acid, lithium potassium sodium salt

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001, September 20 to 2001 October 11, 2001

Reliability:

1 (reliable without restriction)

Data source

Reference

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted February 24, 1987

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 31, 1992.

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

440-770-9

EC Name:

-

Cas Number:

371921-63-0

Molecular formula:

C38 H29 Cl2 N5 O12 S4 .x K .x Li .x Na

IUPAC Name:

3,10-diamino-2-{[6-(4-tert-butylbenzenesulfonamido)naphthalen-2-yl]sulfonyl}-6,13-dichloro-5,12-dioxa-7,14-diazapentacene-4,11-disulfonic acid lithium hydride potassium hydride sodium hydride

Test material form:

solid

Test animals

Species:

rat

Strain:

other: HanBrl: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST SYSTEM
Source: RCC Ltd; Biotechnology and Animal Breeding Division; CH-4414 Füllinsdod / Switzerland
Number of animals per group: 5 males or 5 females
Total number of animals: 5 males and 5 females
Age when treated: Males: 9 weeks; Females: 11 weeks
ldentification: Unique cage card and corresponding color-coded spots on the tail.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

HUSBANDRY
Room number: E16
Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with target ranges for temperature 22 + 3"C and for relative humidity between 30-70%. 12 hours fluorescent light/12 hours
dark (light period between 6.00 and 18.00), music during the light period.

Accommodation: ln groups of three per sex in Makrolon type-4 cages with wire mesh tops and standardized softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).

Diet: Pelleted standard Provimi Kliba 3433 raVmouse maintenance diet, batch no.73/01 (Provimi Kliba AG, CH-4303 Kaiseraugst Switzerland) ad libitum. Results of analyses for contaminants are archived at RCC Ltd, Itingen.

Water: Community tap-water, from ltingen ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, ltingen.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: back
- % coverage: 10
- Type of wrap if used: The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels.
- Time after start of exposure: 24h after the application

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): 0.33 g test item /mL vehicle
- Constant volume or concentration used: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 6mL (vehicle + test item)/kg
- Concentration (if solution): 0.33 g test item /mL vehicle
- Lot/batch no. (if required): Supplier: Fluka Chemie AG, CH-9471 Buchs (Batch: 42471811 42701)
- Purity: not specified

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
Mortality/viability: Daily during acclimatization and twice daily during days 1-15.

- Necropsy of survivors performed: yes
At the end of the observation period all animals were sacrificed by intraperitoneal injection of NARCOREN (Rhone Merieux GmbH, D-88471 Laupheim) at a dose of at least 2.0 ml/kg body weight (equivalent to at least 320 mg sodium pentobarbitone/kg body weight). The animals were examined macroscopically and all abnormalities recorded. Thereafter, they were discarded.

- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Body weights: On test days 1 (pre-administration), 8 and 15.
Clinical signs: Daily during acclimatization and at least four times on test day 1 after the test item administration. Once daily during days 2-15. All abnormalities were recorded.

Statistics:

No statistical analysis was used.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No deaths occurred during the study.

Clinical signs:

Blue skin and test item residuals were noted in all males on test day two until test day six.
Test item residuals were noted in all females on test day two until test day six and blue skin was observed until test day twelve.

Body weight:

The body weight of the animals was within the range commonly recorded for this strain and age.

Gross pathology:

No macroscopic findings were observed at necropsy.

Applicant's summary and conclusion

Conclusions:

The dermal LDso (rat) of BLUE GS 5664.80 is > 2000 mg/kg bw in test performed according to OECD TG 402 and following GLP.

Executive summary:

The acute dermal toxicity of BLUE GS 5664.80 has been determined in the current study performed following OECD TG 402 according to GLP.

A group of five male and five female HanBrl: WIST (SPF) rats was treated with BLUE GS 5664.80 at 2000 mg/kg by dermal application. The test item was diluted in vehicle (PEG 300) at a concentration of 0.33 g/ml and administered at a volume of 6 ml/kg. The animals were examined for clinical signs four times during test day 1 and once daily during test days 2-15.

Mortality/viability was recorded together with clinical signs at the same time intervals on test day 1. During test days 2-15 it was recorded two times a day. Body weights were recorded on day 1 prior to administration and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study.

Blue skin and test item residuals were noted in all males on test day two until test day six.

Test item residuals were noted in all females on test day two until test day six and blue skin was observed until test day twelve.

The body weight of the animals was within the range commonly recorded for this strain and age.

No macroscopic findings were observed at necropsy.

The median lethal dose of BLUE GS 5664.80 after single dermal administration to rats of both sexes, observed over a period of 14 days is:

LDso (rat): > 2000 mg/kg bw