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

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with GLP. No test guidance is referenced in the study report. The methodology used is equivalent to OECD Guideline 404 (Acute Dermal Irritation / Corrosion).

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 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

No details of test guidance referenced in study report. The methodology used is equivalent to OECD Guideline 404 (Acute Dermal Irritation / Corrosion).

GLP compliance:

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: NBY 405/74) was received as a dark brown powder. It was sent for testing from ICI Specialties, Blackley, Manchester, UK. A certificate of analysis (reference: 9108127, dated 8' August 1991) stated that the test sample contained 71-6% w/w Substance H112339, From information supplied hy the sponsor the test sample was used within the stated expiry date. The test sample was given the CTL reference number Y07719/001 and was tested as supplied (moistened to a paste with a small amount of deionised water).

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: MelTor Rabbits
- Age at study initiation: -
- Weight at study initiation: 3699-4034g
- Housing: The rabbits were housed individually in anodised aluminium sheet cages (55.2cm width x 68.9cm depth x 45.7cm height). The floor of each cage was made of perforated aluminium sheet and the door was made of stainless steel rods.
- Diet (e.g. ad libitum): Labsure CRB Rabbit Diet, ad libitum
- Water (e.g. ad libitum): via an automatic system, ad libitum
- Acclimation period: minmum 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17°C (± 2⁰C)
- Humidity (%): 55 (±15%)
- Air changes (per hr): 20-30
- Photoperiod (hrs dark / hrs light): controlled by a time switch to give alternate periods of artificial light and 12 hours of darkness

Test system

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Remarks:

deionised

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500mg

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5ml water

Duration of treatment / exposure:

Exposure 4 hours

Observation period:

3 days

Number of animals:

3

Details on study design:

Three female rabbits were used for this study. Approximately twenty-four hours before application of the test sample, the hair was removed with a pair of veterinary clippers from an area approximately 7cm x 13cm on the left flank of each animal. Each animal was given a number, unique within the study, which was written on the inner side of one ear, using a black waterproof marker-pen, and displayed on the cage card. The bodyweight of each animal was recorded at the start of the study.

The test sample (approximately 500mg) was moistened with a small amount (0.5ml) of deionised water and was applied to the test site (approximate size 2.5cm x 2.5cm) on the left flank of each rabbit using a metal spatula.

The treated areas were covered with pieces of surgical gauze (approximate size 2.5cm x 2.5cm; 8 ply) each of which was secured by two crossed strips of surgical tape (approximate size 1cm x 8cm). These were covered by a piece of impermeable rubber sheeting (approximate size 35cm x 13cm) wrapped once around the trunk of the animal and secured with adhesive impermeable polyethylene tape (7.5cm wide).
The dressings were left in position for approximately four hours. After this tine each dressing was carefully cut (using blunt-tipped scissors), removed and discarded. The application sites were gently cleansed free of any residual test sample using clean swabs of absorbent cotton wool soaked in clean warm water and were then dried gently with clean tissue paper. The application sites were identified using a black waterproof marker-pen.

On day three of the study the rabbits were humanely killed by intravenous injection of pentobarbitone (EUTHATAL, RMB Animal Health) and the skin was removed and examined histologically for evidence of irritation or necrosis. A sample of skin from the application site, as well as a sample from the apposite (control) flank, was removed from each rabbit and fixed in Bouins solution. The tissue samples were then processed, embedded in paraffin wax, sectioned at 5µm, stained with haematoxylin and easin and examined under the light microscope.

The Draize scale was used to assess the degree of erythema and oedema at the application sites approximately 30-60 minutes, 1, 2 and 3 days after removal of the dressings. Any other signs of skin irritation were also noted.
The mean oedema scores were calculated for each animal by adding together the oedema scores at the 1, 2 and 3-day readings and dividing the total by three. It was not possible to calculate a mean erythema score as the test substance stained and obscured the application sites.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

2.3

Max. score:

4

Reversibility:

no data

Remarks on result:

other: There was no evidence, of widespread necrosis, therefore it is likely that the effects were reversible, however no further information on this is available.

Results 2

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

no data

Remarks on result:

other: There was no evidence, of widespread necrosis, therefore it is likely that the effects were reversible, however no further information on this is available.

Results 3

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

2.7

Max. score:

4

Reversibility:

no data

Remarks on result:

other: There was no evidence, of widespread necrosis, therefore it is likely that the effects were reversible, however no further information on this is available.

Irritant / corrosive response data:

Following a single four hour' application of the undiluted test sample, the application sites of all three animals were stained red. This staining prevented assessment of erythema but did not prevent assessment of oedema.

Very slight oedema was observed in one rabbit and very slight to moderate oedema was observed in the remaining two animals up to 3 days after application of the test sample. In addition, wrinkling of the skin was observed in two animals.

These animals were withdrawn from the study on day three, humanely killed and skin samples submitted for histopathological examination,

Two of the three test sections showed slight multifocal or diffuse acanthosis, slight or moderate multifocal or diffuse inflammatory cell infiltration and slight multifocal parakeratosis/surface debris. Minimal focal epithelial necrosis was also present in one test site. The skin section from the remaining test site showed minimal multifocal inflammatory cell infiltration. Control sections from all animals showed minimal multifocal inflammatory-cell infiltration.

The histopathological findings indicate an irritant effect of slight to moderate intensity in two animals, There was no evidence, of widespread necrosis, therefore it is likely that the effects were reversible.

Other effects:

The animals were withdrawn from the study on day three, humanely killed and skin samples submitted for histopathological examination.

Applicant's summary and conclusion

Interpretation of results:

moderately irritating

Remarks:

Migrated information to rabbit skin Criteria used for interpretation of results: EU

Conclusions:

Three female rabbits each received a single four hour application of the test sample to the shorn flank. The animals were assessed for any signs of irritation for up to 3 days after application.

The test sample stained the application sites of all animals and prevented assessment of erythema. On day 3 of the study, animals were humanely . killed and samples of skin were taken and examined histopathologically for signs of skin irritation.

Substance HI 12339 was a moderate irritant to rabbit skin following a single four hour application.

Executive summary:

Substance H112339 was a moderate irritant to rabbit skin following a single four hour application. The substance is classified on this basis as follows:

Dangerous Substance Directive (67/548/EEC): Xi – R38

CLP Regulation (EC No 1272/2008) Skin Irrit. 2 - H315