Anthra[2,1,9-mna]naphth[2,3-h]acridine-5,10,15(16H)-trione

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1981/1982

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

data were already available

Test material

Test material information

Constituent 1

Reference substance name:

Anthra[2,1,9-mna]naphth[2,3-h]acridine-5,10,15(16H)-trione

EC Number:

221-897-7

EC Name:

Anthra[2,1,9-mna]naphth[2,3-h]acridine-5,10,15(16H)-trione

Cas Number:

3271-76-9

Molecular formula:

C31H15NO3

IUPAC Name:

anthra[2,1,9-mna]naphth[2,3-h]acridine-5,10,15(16H)-trione

Test material form:

solid: particulate/powder

Details on test material:

Vat Green 3

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Animal Breeding Unit, Imperial Chemical Industries PLC, Pharmaceuticals Division, Alderley Park, Macclesfield, Cheshire
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 4 to 7 weeks
- Weight at study initiation: 270 to 450 g
- Housing: The guinea pigs were housed in suspended stainless steel mesh cages (370 mm length X 320 mm width X 200 mm height). The floor and back of each cage were of 12 mm square stainless steel mesh, the sides were of solid stainless steel and the front was of polycarbonate (Makrolon). The animals were housed individually with 2 animals /cage.
- Diet (ad libitum): RGP Guinea pig Diet
- Water (ad libitum): tap water ad libitum via an automatic water system
- Acclimation period: at least one week
- Indication of any skin lesions: none

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23
- Humidity (%): 50 to 60
- Air changes (per hr): 10 to 20
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

water

Concentration / amount:

1% / 0.1 mL

Day(s)/duration:

Day 1

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

75% / 0.2 - 0.3 mL

Day(s)/duration:

Day 8 / 48 h

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

75% and 25% / 0.2 - 0.3 mL

Day(s)/duration:

Day 22 / 24 h

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Number of animals in test group: 20 Number of animals in negative control group: 10

Details on study design:

A. INDUCTION EXPOSURE
The hair was removed of an area, approximately 50 mm x 50 mm on a scapular region of each animal and a row of three injections. (0.1 mL each) was made of each side of the mid-line.
The injections were:
- (i) Freund’s complete adjuvant plus distilled water in the ratio 1:1
(ii) A suspension of the test sample indistilled water.
(iii) A suspension of the Test Sample in distilled water, emulsified with freund’s adjuvant.
ONE WEEK later, the scapular area was clipped again and treated with topical application of the Test Sample as a 75 % (w/v) suspension in distilled water.
Approximately 0.2-0.3 mL of the suspension was applied on filter paper, which was held in place by a piece of surgical tape. The tape was covered by a strip of adhesive bandage around which was wrapped once only, self adhesive PVC tape. This occlusive dressing was kept in place for 48 hours.

B. CHALLENGE EXPOSURE

TWO WEEKS after the topical inductions, an area approximately 150 mm x 50mm, on both flanks of each animal was clipped free on hair using veterinary clippers.
Occlusive dressings were prepared.
Approximately 0.2 -0.3 mL of the suspension of the test sample 75% and 25% in distilled water was placed on the filter paper and the dressing was place on the shorn flanks. The dressing was then covered with a strip of adhesive bandage.
After 24 hours, the occlusive dressings were removed, and discarded.
The skin at the site of application was then cleansed of any residual Test Sample by using clean swabs of cotton wool soaked in clean warm tap water and was then dried gently with clean tissue paper.
24 and 48 hours following decontamination, reactions were quantified using a four point scale and the number of positive responses were recorded.

Results and discussion

In vivo (non-LLNA)

Results

Clinical observations:

Skin discolouration prevented the evaluation of skin reddening - histopathological evaluation reveled no skin sensitising potential

Remarks on result:

no indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Histopathological examination of the skin indicated that the test substance was not sensitive to guinea pig skin

Executive summary:

The skin sensitising properties of the test substance were assessed using the maximisation test of Magnusson and Kligman (1970).

Thirty Dunkin Hartley albino female guinea pigs, twenty test and test control were used for the test.

Three main procedures were involved; intradermal injection of a 1% suspension of the test sample in distilled water, topical application of a 75% suspension of the test sample in distilled water and challenge with a 75% and 25% suspension of the test sample in distilled water.

The challenge sites were examined twenty-four and forty-eight hours following the removal of the dressings and any erythematous reactions were quantified at those times. In this study however, the skin assessments were impeded due to the staining by the test sample and samples of test control skin of each guinea pig were submitted for histopathological examination.

Following the 25% and 75% challenge, no erythematous reactions could be seen at twenty-four or fourty-eight hours due to the staining of the skin by the sample.

Histopathological examination of the skin indicated that the test substance was not sensitive to guinea pig skin.