Registration Dossier

Diss Factsheets

Administrative data

Description of key information

FAT 65004 is considered as non-sensitiser.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation, other
Remarks:
SKIN PHOTOALLERGENICITY
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
Gunea Pig were treated with test substances and effects observed post epidermal challenge applications followed by UV-B and UV-A irradiation or followed by UV-A irradiation.
GLP compliance:
no
Type of study:
Draize test
Specific details on test material used for the study:
Test material: FAT 65004/C
Identification: Uvitex SK (DCT No. 90043)
Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
The test was performed on 10 male and 10 female guinea pigs per group weighing between 270 - 370 g (Approx. 10 weeks old). The animals were housed individually in Macrolon cages (type 3) , assigned to the different groups by means of random numbers generated by the random number generator incorporated in the Hewlett-Packard desk computer, identified with individual ear tags, kept at a constant room temperature of 20±1 C°, at a relative humidity of 55 ±5 % and on a 14 hours
light cycle day. The animals received ad libitum standard guinea pigs pellets - NAFAG No. 830, Gossau SG - supplemented with fresh carrots and fresh water.
Route:
epicutaneous, open
Vehicle:
other: DAE433
Concentration / amount:
0.1 % suspension of FAT 65004/C in 80 % DAE433
Day(s)/duration:
4 Times a week for three weeks the animals were treated
No.:
#1
Route:
epicutaneous, open
Vehicle:
other: DAE433
Concentration / amount:
0.1 % suspension of FAT 65004/C in 80 % DAE433
Day(s)/duration:
3 days
Adequacy of challenge:
not specified
No.:
#2
Route:
epicutaneous, open
Vehicle:
other: DAE433
Concentration / amount:
0.1 % suspension of FAT 65'004/C in 80 % DAE433
Day(s)/duration:
3
Adequacy of challenge:
not specified
No. of animals per dose:
10 male and 10 female
Details on study design:
Method:
Induction:
4 days before starting the induction, the animals were shaven on the neck and chemically depilated. During the whole induction period the animals were only shaven (Aesculap , 1/20 mm). 4 Times a week for three weeks the animals were treated by epidermal open exposure with a 0.1 % suspension of FAT 65004/C in 80 % DAE433 (0.1 ml per application; 2x2 cm application area on the animals' neck).
One hour after application the animals were irradiate for 14 minutes during the first induction week (UV-A, visible), 10 minutes during the second and third week (UV-B, UV-A, visible).
During the irradiation process the animals were immobilized in a special animal holder. In order to protect the skin surrounding of the irradiated area, this part was covered with a lightproof bandage.
24 hours after each of the first four induction applications, the skin reactions were evaluated according the Draize Scoring Scale. The reactions during the second and third induction week were not recorded.
A total of 4 adjuvant injections were made to the four corners of the application site on Monday and Wednesday of the second and third induction week (0.1 ml suspension of adjuvant complete
Freund and physiological saline per injection).

Challenge I:
12 days after the last induction irradiation, the animals were shaven on the back and the skin was chemically depilated (5 min.). 4 days later the animals were treated by epidermal open exposure for three days as during the induction period. One hour after each application the animals were irradiated for 7 minutes with a sub-erythematogenic dose of UV-B, UV-A and visible light (lamp filtered with Schott filter WG 280,3 mm).

Challenge II:
A second challenge was done after a test period of 14 days. The substance was applied again three times on the shaven skin of the animals back. The test site was irradiated after each application for 15 minutes with a sub-erythematogenic dose of UV-A and visible light (lamp filtered with Schott WG 320, 3 mm).

24 hours after each application of both challenges, the skin reactions were evaluated according the Draize Scoring Scale. Every animal with one or more score values higher at the end of the two challenge periods than that obtained at the end of the first induction week was termed positive. The number of positive animals in the test group was compared with the number of animals in the control group treated with the vehicle alone.


Challenge controls:
DAE4 33 control
Positive control substance(s):
not specified
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
test chemical
No. with + reactions:
1
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation

The individual erythema reactions were evaluated evaluated 24 hours after the last irradiation of either the induction or challenge I and challenge II periods.

Only one animal of the last group showed a reaction at the end of challenge period II. One animal (No. 863) died during the rest period after the induction procedure, unrelated to the treatment.

On the basis of these results, a photoallergenic potential of FAT 65 004/C in guinea-pigs can be excluded.

RESULTS:

   No of positive animals/ No of treated animals

Challenge I

DAE433 Control group

FAT65004/C

 

0/20

0/19

Challenge II

DAE433 Control group

FAT65004/C

 

0/20

1/19

Individual erythema reaction score 24 hours after the last irradiation of the induction, challenge I and challenge II:

Control Group  Animal number
DAE433 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840
Induction 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Challenge I 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Challenge II 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Test Group  Animal number
FAT 65004/C 861 862 863 864 865 866 867 868 869 870 871 872 873 874 875 876 877 878 879 880
Induction 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Challenge I 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Challenge II 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Interpretation of results:
GHS criteria not met
Conclusions:
On the basis of these results, a photoallergenic potential of FAT 65004/C in guinea-pigs can be excluded.
Executive summary:

Under the experimental condition employed, no differences between the test group and the vehicle-treated controls were seen, after epidermal challenge applications either followed by UV-B and UV-A

irradiation or followed by UV-A irradiation alone.

FAT 65004/C was found to be devoid of a photoallergenic potential in albino guinea-pigs.

Endpoint:
skin sensitisation, other
Remarks:
SKIN PHOTOTOXICITY
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
Mice were treated with test substances and effects observed post epidermal applications followed by UV- irradiation.
GLP compliance:
no
Type of study:
Draize test
Specific details on test material used for the study:
Test material: FAT 65004/C
Identification: Uvitex SK (DCT No. 90043)
Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
Strain: Mouse, SC: hairless - 1
Breeder: Bomholtgard, Denmark
Acclimatisation period: 13 days

The test was performed on 3 male and 3 female mice per group (SC: hairless -1, outbred mice) weighing 25 - 33 g (2.6 to 3 months old). They were housed in Macrolon cages (size 2) in groups of 3 individuals per cage, assigned to the different
groups by means of random numbers generated by the random number generator incorporated in the Hewlett-Packard desk computer, identified with colour markers on the tail, kept at a constant room temperature of 22 ±1°C, at a relative
humidity of 55 ±5 % and on a 14 hours light cycle day. The animals received ad libitum standard mouse pellets No. 890 - Nafag Gossau SG - and water.

Route:
epicutaneous, open
Vehicle:
other: DAE433
Concentration / amount:
Suspension of 0.3 %, 1 % and 3 % FAT 65004/C in 100 % DAE433
Day(s)/duration:
72 hrs
No. of animals per dose:
3 male and 3 female
Details on study design:
The test was performed on 3 male and 3 female mice per group (SC: hairless -1, outbred mice) weighing 25 - 33 g (2.6 to 3 months old). They were housed in Macrolon cages (size 2) in groups of 3 individuals per cage, assigned to the different groups by means of random numbers generated by the random number generator incorporated in the Hewlett-Packard desk computer, identified with colour markers on the tail, kept at a constant room temperature of 22 ±1 C°, at a relative humidity of 55 ±5 % and on a 14 hours light cycle day. The animals received ad libitum standard mouse pellets No. 890 - Nafag Gossau SG - and water.
Suspension of 0.3 %, 1 % and 3 % FAT 65004/C in 100 % DAE433 were prepared. Aliquots of 0.05 ml of each concentration were applied evenly with a pipet onto the skin of both flanks of the mice. For each concentration level a separate group of animal was employed. Negative and positive control groups treated either which the vehicle or a solution of 0.1 % 8-methoxypsoralene respectively were included in the test system.
Half an hour after application the animal was fixed in front of the light source in an animal holder at a distance of 5.6 cm from the lamp box and a circular skin area of 1 cm in diameter of one side of the mouse was exposed to UV-light for a duration of 2 minutes. The contralateral side, not exposed to light, served as the control.
The skin reactions were evaluated 24, 48 and 72 h after the irradiation according to the Draize Scoring system for primary irritation reactions.
Positive control substance(s):
yes
Remarks:
0.1 % 8-methoxypsoralene (Sigma), 8-MOP.
Key result
Reading:
other: phototoxicity
Hours after challenge:
72
Group:
test chemical
Dose level:
0.3, 1.0 and 3%
No. with + reactions:
0
Total no. in group:
6
Remarks on result:
other: No indication of irritation/toxicity

No adverse skin reaction was seen when FAT 65004/C was applied topically to hairless mice followed by UV-irradiation.

Slight erythema and moderate oedema reactions were seen 24 h after appplication of 0.1 % 8-methoxypsoralene topically to hairless mice followed by UV-irradiation. Fresh necrosis

surrounded by erythema was observed 48 and 72 h after irradiation in the 0.1 % 8-methoxypsoralene group.

Number of positive animals per group:

24 h after irradiation:

No. of positive animals/
No. of treated animals/ 		 No. of positive animals/
No. of treated animals/ 
IRRIDATED FLANK NON-IRRIDATED FLANK
Vehicle control 0/6 0/6
0.1% 8-MOP 0/6 0/6
0.3 % FAT65004/C 0/6 0/6
1.0 % FAT65004/C 0/6 0/6
3.0 % FAT65004/C 0/6 0/6

48 h after irradiation

No. of positive animals/
No. of treated animals/ 		 No. of positive animals/
No. of treated animals/ 
IRRIDATED FLANK NON-IRRIDATED FLANK
Vehicle control 0/6 0/6
0.1% 8-MOP 0/6 0/6
0.3 % FAT65004/C 0/6 0/6
1.0 % FAT65004/C 0/6 0/6
3.0 % FAT65004/C 0/6 0/6

72 h after irradiation:

No. of positive animals/
No. of treated animals/ 		 No. of positive animals/
No. of treated animals/ 
IRRIDATED FLANK NON-IRRIDATED FLANK
Vehicle control 0/6 0/6
0.1% 8-MOP 0/6 0/6
0.3 % FAT65004/C 0/6 0/6
1.0 % FAT65004/C 0/6 0/6
3.0 % FAT65004/C 0/6 0/6

Every animal with a defined erythema or edema was evaluated as a positive animal.

Interpretation of results:
GHS criteria not met
Conclusions:
FAT 65004/C was found to be devoid of a phototoxic potency in hairless mice.

Executive summary:

Under the experimental conditions employed, FAT 65004/C was found to cause neither erythema nor edema in hairless mice when applied topically followed by UV irradiation.

FAT 65004/C was found to be devoid of a phototoxic potency in hairless mice.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

In a key study conducted to assess the phototoxicity 3 male and 3 female mice per group were treated by epidermal open exposure with a 0.1 % suspension of FAT 65004/C in DAE433and half an hour after application the animal was fixed in front of the light source in an animal holder at a distance of 5.6 cm from the lamp box and a circular skin area of 1 cm in diameter of one side of the mouse was exposed to UV-light for a duration of 2 minutes. The contralateral side, not exposed to light, served as the control.The skin reactions were evaluated 24, 48 and 72 h after the irradiation according to the Draize Scoring system for primary irritation reactions. No adverse skin reaction was seen when FAT 65004/C was applied topically to hairless mice followed by UV-irradiation.

Irradiation using UV & visible light can be viewed as an enhanced methodology to assess the toxicity and allergic reactions of the degraded products or metabolites.

In the above mentioned photo-toxicity test no adverse skin reaction was seen when FAT 65004/C was applied topically. Which indicate the non-toxic properties of degraded products or metabolites formed after irradiation on mice skin.

In the next part of the experiment, Guinea pigs were treated by epidermal open exposure with a 0.1 % suspension of FAT 65004/C in DAE433 and subjected to UV light to assess the allergic potential.

As induction the animals 4 times a week for three weeks the animals were treated by epidermal open exposure with a 0.1 % suspension of FAT 65004/C in 80 % DAE433 (0.1 ml per application; 2x2 cm application area on the animals' neck). One hour after application the animals were irradiate for 14 minutes during the first induction week (UV-A, visible), 10 minutes during the second and third week (UV-B, UV-A, visible). During the irradiation process the animals were immobilized in a special animal holder. In order to protect the skin surrounding of the irradiated area, this part was covered with a lightproof bandage. 24 hours after each of the first four induction applications, the skin reactions were evaluated according the Draize Scoring Scale. The reactions during the second and third induction week were not recorded. A total of 4 adjuvant injections were made to the four corners of the application site on Monday and Wednesday of the second and third induction week (0.1 ml suspension of adjuvant complete Freund and physiological saline per injection).

1st challenge dose was initiated 12 days after the last induction irradiation. The animals were shaven on the back and the skin was chemically depilated (5 min.). 4 days later the animals were treated by epidermal open exposure for three days as during the induction period. One hour after each application the animals were irradiated for 7 minutes with a sub-erythematogenic dose of UV-B, UV-A and visible light (lamp filtered with Schott filter WG 280,3 mm).

A 2nd challenge was done after a test period of 14 days. The substance was applied again three times on the shaven skin of the animals back. The test site was irradiated after each application for 15 minutes with a sub-erythematogenic dose of UV-A and visible light (lamp filtered with Schott WG 320, 3 mm).

24 hours after each application of both challenges, the skin reactions were evaluated according the Draize Scoring Scale. Every animal with one or more score values higher at the end of the two challenge periods than that obtained at the end of the first induction week was termed positive. The number of positive animals in the test group was compared with the number of animals in the control group treated with the vehicle alone.

Under the experimental condition employed, no differences between the test group and the vehicle-treated controls were seen, after epidermal challenge applications either followed by UV-B and UV-A irradiation or followed by UV-A irradiation alone. This indicates the non-allergic character of the test substance FAT 65004 and also of the degraded products or metabolites that might have formed during the Irradiation process.

Hence, FAT 65004 can be considered as non-sensitizing to guinea pigs.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the available data from selected key studies, FAT 65004 does not warrant classification for skin sensitisation under the CLP (1272/2008) Regulation.