Reaction mass of 5,5'-{(phenylmethanediyl)bis[benzene-4,1-diyl-diazene-2,1-diyl]}bis{1-[3-(dimethylamino)propyl]-4-methyl-6-oxo-3-(pyridinium-1-yl)-1,6-dihydropyridin-2-olate} hydrochloride and 5,5’-[3,4’-(phenylmethanediyl)diphenylene]bis(diazene-2,1-diyl)bis{1-[3-(dimethylamino)propyl]-4-methyl-6-oxo-3-(pyridinium-1-yl)-1,6-dihydropyridin-2-olate} hydrochloride

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2007-09-24 to 2008-01-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations, conducted with the analogue substance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

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:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

analogue substance (refer to IUCLID chapter 13)

IUPAC Name:

analogue substance (refer to IUCLID chapter 13)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services, Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
- Age at study initiation: females 11 weeks, males 8 weeks
- Weight at study initiation: females 197.4 - 213.5 g, males 241 - 261.2 g
- Fasting period before study: no
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard rat/mouse maintenance diet ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark

IN-LIFE DATES: From: 2007-09-25 To: 2007-10-16

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: backs of the animals
- % coverage: ca. no data
- Type of wrap if used: The semi-occlusive dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.

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

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 6 ml/kg bw
- Concentration (if solution): 333.3 mg/l
- Constant volume or concentration used: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 6 ml/kg bw
- Purity: Purified water prepared at RCC Ltd (deionised water which was processed and treated by the PURELAB Option-R unit. This latter links four purification technologies: reverse osmosis, adsorption, ion-exchange and photo oxidation).

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality / Viability: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.
Clinical signs: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15. All abnormalities were recorded.
Weighing: On test days 1 (prior to administration), 8 and 15.
Local Signs: Once daily during days 2-15. All abnormalities will be recorded.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

No statistical analysis was used.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Analogue substance.

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: Signs of toxicity related to dose levels: No systemic signs of toxicity were observed. Local toxicity: At removal of the application patch the local skin reactions were not assessable in all animals due to a yellow staining produced by the test item. The

Gross pathology:

Effects on organs: No macroscopic findings were observed at necropsy.

Other findings:

- Organ weights: no data
- Histopathology: no data
- Potential target organs: not applicable
- Other observations:

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The median lethal dose of the test item after single dermal administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat) > 2000 mg/kg body weight. Based on the proposed read across approach, the LD50 of the substance registered can be assumed to be in the same range as for the analogue substance.

Executive summary:

The acute dermal toxicity to rats of the analogue substance was being investigated following the testing protocol as given in OECD guideline 402.

Five male and five female rats were treated with the test item at 2000 mg/kg by dermal application. The test item was diluted in vehicle (purified water) at a concentration of 0.33 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. Body weights were recorded. All animals were necropsied and examined macroscopically.

No deaths occurred during the study.

At removal of the application patch the local skin reactions were not assessable in all animals due to a yellow staining produced by the test item. The assessment was prevented up to day 6 in the females and day 7 in the males. The staining was present from day 7 in the females and on day 8 in the males.When assessable, a slight local erythema was noted in one male from days 8 to 9 and in all females from day 7 to 9 or 10. Slight scaling was present from day males and from days 7 or 8 up to day 10 in all females. Slight crusts were noted in one female from day 7 to 13.

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 the test item after single dermal administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat) > 2000 mg/kg body weight.

Given the applicability of the proposed read across approach (see IUCLID chapter 13), the respective LD50 values of the substance registered can safely be assumed to be in the same range as for the analogue substance, as (1) the molecules of the substance registered and the analogue substance show to a large extent structural similarities, and (2) furthermore the molecular weights of the analogue substance and the substance registered only differ by 2.5%.