Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 1991 - November 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with GLP. No Guideline specified, although the methodology is equivalent to OECD Guideline 402 (Acute Dermal Toxicity).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Principles of method if other than guideline:

No details of guideline in the study report although the methodology is equivalent to OECD Guideline 402 (Acute Dermal Toxicity).

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

EC Number:

413-550-5

EC Name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Cas Number:

142068-96-0

Molecular formula:

Hill formula: C53H28Cl2N14Na8O26S8

IUPAC Name:

octasodium 5-[(4-chloro-6-{[4-({[4-chloro-6-({7-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-8-hydroxy-3,6-disulfonatonaphthalen-1-yl}amino)-1,3,5-triazin-2-yl]amino}methyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-3-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate

Details on test material:

The test sample (reference: NBY4Q5/74) was received as a dark brown powder. It was sent for testing from ICI Specialties, Blackley, Manchester, UK. A certificate of analysis (reference: 9103127 and dated 8 August 1991) stated that the test sample contained 71.6% (w/w) substance H112339 and 10.5% (w/w) moisture. From information supplied by the sponsor the test sample was used within the stated expiry date.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Barriered Animal Breeding Unit, ICI Pharmaceuticals
- Age at study initiation: young adults
- Weight at study initiation: Male - 239-268g, Females - 183-215g
- Fasting period before study: No
- Housing: The rats were housed in suspended cages (37,5cm length x 32,5cm width x 23.0cm height). The cages were constructed of stainless steel, two sides being sheet and the remainder 14 standard wire gauge mesh. The animals were housed individually.
- Diet (e.g. ad libitum): Porton Combined Diet, ad libitum
- Water (e.g. ad libitum): via an automatic system, ad libitum
- Acclimation period: minimum of 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 and 23°C
- Humidity (%): 55±15%.
- Air changes (per hr):25-30
- Photoperiod (hrs dark / hrs light): controlled by a time switch to give alternate periods of 12 hours of artificial light and 12 hours of darkness

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 10cm x 5cm
- % coverage: -
- Type of wrap if used: gauze patch covered by a patch of plastic film held in position using adhesive bandage. This was secured by two pieces of PVC tape wrapped around the animal.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cleansed free of any residual test sample using clean swabs of absorbent cotton wool soaked in clean warm water and methylated spirits
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000mg/l
- Concentration (if solution): -
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.4ml
- Concentration (if solution): -
- Lot/batch no. (if required): -
- Purity: -

Duration of exposure:

24 hours

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

Male - 5
Female - 5

Control animals:

no

Details on study design:

Five male and five female rats were used for this study. Sixteen to thirty-two hours before application of the test sample, the hair was removed with a pair of veterinary clippers from an area, approximately 10cm x 5cmr on the dorso-lumbar region of each animal. Each animal was given a number, unique within the study, using an ear-punch.

The main study was preceded by a preliminary study (speculative dermal study) in which a dose-level of 2000mg/kg was tested on a small group of animals. Based on information from this study the dose level for the main study was selected as 2000mg/kg. The dose level was adjusted to account for the moisture content, therefore a dose of 2230mg/kg test substance as supplied was applied which is approximately equal to 2000mg/kg of the test material excluding water.

The appropriate amount of the test sample was weighed out onto a .plastic weighing boat and made into a paste by adding a small amount (approximately 0.4ml) of deionised water. The amount applied was calculated for each animal according to its weight at the time of dosing. The paste, covered by a gauze patch (approximately 4cm x 6cm x 4-ply) was applied to the shorn backs of the animals and was kept in contact with the skin for approximately 24 hours using occlusive dressings. The gauze patch was covered by a patch of plastic film (7.5cm x 5cm) and was held in position using adhesive bandage (25cm x 5cm), This was secured by two pieces of PVC tape (approximately 2.5cm x 20cm) wrapped around the animal.

At the end of the 24-hour contact period, the dressings were carefully cut, using blunt-tipped scissors, removed and discarded. The skin, at the site of application, was cleansed free of any residual test sample using clean swabs of absorbent cotton wool soaked in clean warm water and methylated spirits and was then dried" gently with clean tissue paper.

The animals were observed for signs of systemic toxicity once between one and four hours after application (only gross abnormalities were noted at this time as the presence of the dressings may have affected the behaviour and movement of the rats) and then once daily for systemic toxicity and skin irritation up to Day 15. The animals were weighed immediately before application of the test sample (day 1), and on day 3, day 4, day 8 and day 15,

At the end of the study, all of the animals were humanely killed by inhalation of excessive levels of halothane BP (FLUGTHANE, ICI Pharmaceuticals) vapour followed by exsanguination and were subjected to a post mortem examination.

Statistics:

Not reported.

Results and discussion

Preliminary study:

The main study was preceded by a preliminary study (speculative dermal study) in which a dose-level of 2000mg/kg was tested on a small group of animals. Based on information from this study the dose level for the main study was selected as 2000mg/kg.

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

None of the animals died.

Clinical signs:

other: There were no signs of toxicity. Slight skin irritation, indicated by oedema, was observed in all animals on day 2 but had disappeared in the majority of animals by day 3.

Gross pathology:

There were no macroscopic abnormalities detected in any of the animals at the post mortem examination

Other findings:

The test substance stained all the application sites red and persisted, in the majority of the animals, throughout the study preventing accurate assessment of irritation.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute dermal median lethal dose of the substance was greater than 2000mg/kg to both male and female rats. There was evidence of slight skin irritation, in all animals, on day 2 only

Executive summary:

Study conducted in compliance with GLP. No guidelines are detailed in the report.

LD50 > 2000 mg/kg body weight. The substance is not classified as harmful by dermal application.