6,18-dihydrodinaphtho[2,3-i:2',3'-i']benzo[1,2-a:4,5-a']dicarbazole-5,7,12,17,19,24-hexone

Endpoint summary

Description of key information

The irritation capacity on the skin and eye of animals was intensivly tested in the past and numerous data with different test substance concentrations are available.

 

Skin irritation:

In total five different studies with different test material concentrations are available. In the latest study FAT 46014/F with a purity of about 36 % was investigated according to OECD 404 in the rabbit. Three animals have been tested for 4 hours on the intact and abraded skin under semiocclusive conditions. Taking the 24 h, 48 h and 72 h readings into account (mean value 0 for erythema and eschar formation and mean value 0 for edema formation) the test substance has to be judged as non-irritating. A second study investigating FAT 46014/E with a purity of 35 % in a pre-guideline study on abraded and intact skin areas under occlusive testing conditions resulted a mean of 1 for erythema/eschar formation and a mean of 1 for edema formation taking the 24 h, 48 h and 72 h readings of the intact skin areas into account. Interestingly abrasion does not lead to further elevation of scores compared with intact skin areas. A third study on FAT 46014/C with a purity of 18 % was performed in a pre-guideline test on the intact and abraded skin under occlusive conditions. Results obtained in 6 animals on intact skin areas resulted in mean values of 0.16 for erythema/eschar formation and 0.16 for edema formation with full recovery after 48 hours. Based on these results the substance has to be judged as not irritating to the skin. Notably, even slightly enhanced effects observed on abraded skin areas returned to normal 3 days after test substance application. In the fourth study FAT 46014/B with a purity of 45 % was investigated in a pre-guideline test in six rabbits. Again the test substance was applied to intact and abraded skin areas under occlusive conditions. Unfortunately, only readings 24 h and 72 h after test substance application have been reported. Nevertheless, the test substance showed no signs of irritation at all (all readings in all animals are zero) and no other effects are reported. Therefore, it is concluded that the test substance did not induce any signs of irritation to the animals. Last but not least a study has been performed on FAT 46014/A with a purity of 45 %. The test material was investigated in a pre-guideline study in six rabbits on intact and abraded skin areas under occlusive conditions. Also in this test only results from the 24 hour and 72 hour reading have been reported. But since all reading on the intact skin have been zero for erythema/eschar formation and edema formation and no other signs of toxicity or irritation effects have been noted the substance is judged to be not-irritating in this test. In summary it can be noted that in general all tests may be used for assessment of the irritation potential to the skin of rabbits since test substance application and terms of testing conditions are comparable or slightly elevated compared to today’s standards (occlusive vs. semiocclusive testing conditions; abrasive vs intact testing conditions). But since data are reported very detailed, some effects of elevated testing conditions might be judged separately. As an overall conclusion it can be noted that up the maximum substance concentration of 45 % the test substance has to be judged as non-irritant according to CLP.

 

Eye irritation:

In total five different studies with different test material concentrations are available. In the latest study FAT 46014/F with a purity of about 36 % was investigated according to OECD 405 in the rabbit. Three animals have been tested by instillation of 0.1 g into one eye while the other eye has been left untreated to serve as internal control. Rinsing of the eye has not been performed (as recommended by the guideline) representing an elevated testing condition. As a result, mean value for the cornea (24, 48 and 72 hour readings) is 1.2, mean value for the iris and mean values for the conjunctiva (24, 48 and 72 hour readings) are 1.9 for redness and 0.66 for chemosis. Taking these results into account the test substance has to be judged as irritating to the eye. A second study investigating FAT 46014/E with a purity of 35 % in a pre-guideline study in six rabbits. Instillation of 0.1 g of the test material into one eye was followed by a rinsing procedure after 30 seconds in 3 animals while in further 3 animals the test substance remained in the eye. Scoring of the animals after 24, 48 and 72 hours resulted in no scores neither for cornea, nor for iris nor for conjunctive (all scores reported, all scores zero). Based on these test results the substance was judged not-irritating to the eye. A third study on FAT 46014/C with a purity of 18 % was performed in a pre-guideline test in six rabbits. Animals have been instilled with 0.1 ml of the test substance in one eye while the other one left untreated and served as internal control. 1 min after test substance instillation eyes have been rinsed with saline to flush remaining test material out of the eyes. While slight scores of conjunctival redness occurred in treated animals at the first reading after one hour all scores had returned to zero after 6 hours. Therefore for the 24, 48 and 72 hour readings all scored have been zero. Based on these test results the substance was judged not-irritating to the eye. In the fourth study FAT 46014/B with a purity of 45 % was performed in a pre-guideline test in six rabbits. Animals have been instilled with 0.1 ml of the test substance in one eye while the other one left untreated and served as internal control. 30 seconds after test substance instillation eyes have been rinsed with saline to flush remaining test material out of the eyes. At the first reading after one day all scores reported have been zero as well as all scores at the 48 and 72 hour readings. No further signs of irritation or other effects on the eye have been reported. Based on these test results the substance was judged not-irritating to the eye. Last but not least a study has been performed on FAT 46014/A with a purity of 45 %. Animals have been instilled with 0.1 mg of the test substance in one eye while the other one left untreated and served as internal control. 30 seconds after test substance instillation eyes have been rinsed with saline to flush remaining test material out of the eyes. At the first reading after one day all scores reported have been zero as well as all scores at the 48 and 72 hour readings. No further signs of irritation or other effects on the eye have been reported. Based on these test results the substance was judged not-irritating to the eye. In summary it can be noted that in general all tests may be used for assessment of the irritation potential to the eye of rabbits (weight of evidence approach). In all cases tests have been conducted either according to OECD guideline 405 or according to the procedure described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO) representing a test standard being used as a guidance to develop OECD Guideline 405 in the late 1980ies. Based on above mentioned results it remains unclear why clear irritating effects have been observed with FAT 46014/F having only a test substance purity of about 36 % while other test sample preparations having much higher test substance concentrations led to no irritation at all (FAT 46014/A and FAT 46014/B). And also results with a comparable purity demonstrated no irritating effects at all (FAT 46014/E). So the only difference is that in all other studies the treated eyes have been flushed with either lukewarm water or physiological saline to remove remaining test material after 30 sec or 1 minute to prevent mechanical damage to the eye which is known to also cause irritating effects. Another option could be that maybe other ingredients present in the coupage of the test material caused those effects, but this remains elusive. In summary, the registrant proposes to judge the test material as non-irritant to the skin and irritant to the eye. Judgement is based on the key study selected for skin irritation and based on a worst case approach taking all available data for eye irritation into account. 

  

Justification for selection of skin irritation / corrosion endpoint:
GLP guideline study

Justification for selection of eye irritation endpoint:
Weight of evidence approach

 

  

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study completion date: 29 February, 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

Specific details on test material used for the study:

Code No. : FAT 46014/F
Batch No. : EN 94071.32
Stability: guaranteed by the sponsor until November 1988
Description: solid
Contents of active ingredient: 35.7 %
Test Article Received: December 6, 1983

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH-4414 Füllinsdorf
- Age at study initiation: 12 - 14 weeks
- Weight at study initiation: 1930-2270 g
- Housing: Animals were housed individually in metal cages.
- Diet (e.g. ad libitum): ad libitum.
- Water (e.g. ad libitum): ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30-70
- Air changes (per hr): approximately 15 air changes/h.
- Photoperiod (hrs dark / hrs light): 12 hours light/day

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 females

Details on study design:

TEST SITE
- Area of exposure: at least 6 cm²

SCORING SYSTEM:
<0.5: non-irritant
0.6 - 3.0: slightly irritant
3.1 - 5.0: irritant
5.1 - 8.0: severely irritant

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0.17

Max. score:

8

Reversibility:

not specified

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

other: Animal #3 had slight erythema at 1 hour observation only

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

other: Animal #3 has slight edema at 1 hour observation only.

Irritant / corrosive response data:

None

Other effects:

None

Scoring of erythema and eschar formation.

Animal No./Sex	1h	24h	48h	72h	days	days
34/F	0	0	0	0
35/F	0	0	0	0
36/F	1	0	0	0
Total	1	0	0	0	= 1	(A)

Scoring of Oedema formation

Animal No./Sex	1h	24h	48h	72h	days	days
34/F	0	0	0	0
35/F	0	0	0	0
36/F	1	0	0	0
Total	1	0	0	0	= 1	(B)

Index of Skin irritation: A1 + B1 = 2 :12 = 0.17

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 46014/F was found to be non-irritant and not corrosive when applied to the rabbit skin.

Executive summary:

The skin irritation potential of FAT 46014/F with purity of 35.7 % was evaluated in a study conducted according to OECD Guideline 404. 3 Female rabbits were used for this purpose. This test was conducted in GLP compliance. The test was carried out on shaved intact skin. Animal #3 had slight erythema and edema at 1 hour observation only, which was resolved within 24 hours. The means scores for 24/48/72 hour observations were 0 for both erythema and edema reactions. Hence, FAT 46014/F can be classified as non-irritant and not corrosive to rabbit skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Study completion date: 12 March, 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Code No. : FAT 46014/F
Batch No. : EN 94071.32
Stability: guaranteed by the sponsor until November 1988
Description: solid
Contents of active ingredient: 35.7 %
Test Article Received: December 6, 1983

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source:Kleintierfarm Madoerin AG, CH-4414 Fuellinsdorf
- Age at study initiation: 12 - 14 weeks
- Weight at study initiation: 1890-2090 g
- Housing: Animals were housed individually in metal cages.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30-70
- Air changes (per hr): approximately 15 air changes/h.
- Photoperiod (hrs dark / hrs light): 12 hours light/day

Vehicle:

unchanged (no vehicle)

Controls:

other: right eye remained untreated and served as a control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g

Duration of treatment / exposure:

one second

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 males

Details on study design:

SCORING SYSTEM:
The ocular reactions were examined 1, 24, 48, and 72 hours after substance instillation and again during the following observation period by means of a slit-lamp. The ocular irritation scores were evaluated according to the scoring system presented in the following table. The irritating and/or corrosive potential of the test article (theoretical range: non irritant - extremely irritant) was assessed on the basis of the maximum irritant effect observed
- in one or several of the ocular tissues (conjunctiva, iris, cornea), observed over the range of observation time
- in one or more animals and taking into account the reversibility of the effect.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1.22

Max. score:

4

Reversibility:

fully reversible within: 7d

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

1.11

Max. score:

2

Reversibility:

fully reversible within: 7d

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

2.56

Max. score:

4

Reversibility:

fully reversible within: 11d

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7d

Irritant / corrosive response data:

None

Other effects:

The animals showed a normal body weight development.

Evaluation of eye reactions

Cornea

Animal No/Sex	31/M	32/M	33/M
After 1 hr	2	2	2
After 24 hr	2	1	2
After 48 hr	1	1	1
After 72 hr	1	1	1
After 7 days	0	0	0
After 11 days	0	0	0

Iris

Animal No/Sex	31/M	32/M	33/M
After 1 hr	1	1	1
After 24 hr	1	2	1
After 48 hr	1	1	1
After 72 hr	1	1	1
After 7 days	0	0	0
After 11 days	0	0	0

Conjuctiva -( redness in bulbar and palpebral membranes)

Animal No/Sex	31/M	32/M	33/M
After 1 hr	2	2	2
After 24 hr	2	3	3
After 48 hr	2	3	3
After 72 hr	1	3	3
After 7 days	0	1	1
After 11 days	0	0	0

Conjuctiva -( chemosis in lids and nictating membranes)

Animal No/Sex	31/M	32/M	33/M
After 1 hr	4	2	3
After 24 hr	2	3	2
After 48 hr	2	3	2
After 72 hr	1	2	1
After 7 days	0	0	0
After 11 days	0	0	0

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

FAT 46014/F is an irritant when applied to the rabbit eye mucosa.

Executive summary:

The eye irritation potential of FAT 46014/ F evaluated in a study conducted according to OECD Guideline 405. 0.1 g of FAT 46014/F was placed into the conjunctival sac of the right eye of each of 3 male rabbits, after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of the test material. The left eye remained untreated and served as a control. The ocular reactions were evaluated 1, 24, 48, and 72 hours after the instillation of FAT 46014/F. The mean scores for corneal opacity, iris, conjunctival redness and chemosis were 1.22, 1.11, 2.56 and 2, respectively. The substance was found to be severely irritant and not corrosive when applied to the rabbit eye mucosa, however the animals had recovered at the end of the observation period of 11 days. Based on the scores obtained and full reversibility, the substance is considered to be eye irritant (category 2).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

In summary, the registrant proposes to judge the test material as non-irritant to the skin and irritant to the eye. Judgement is based on the key study selected for skin irritation and based on a worst case approach taking all data available for eye irritation into account.