Registration Dossier

Administrative data

Description of key information

In a GLP-compliant sensitivity study using the Maximization-Test, performed according to OECD guideline 406 and EU method B.6, guinea pigs were treated with FAT 40400/A. Twenty animals were treated with the test substance and ten animals with only the vehicle (physiological saline). The concentrations of the test substance used in the main study were determined by the results of the preliminary study. The intradermal induction of sensitization in the test group was performed using 5 % of the test substance in both the vehicle and adjuvant/vehicle mixture. One week later this was boosted by the topical application of the test substance at 15 % concentration over the injection sites. Animals of the control group were treated in the same manner but only the selected vehicle was used. Two weeks after the second induction all animals were challenged by topical application of the test substance at 10 % concentration.


Two out of 20 animals showed a positive erythema reaction at the 24 and 48 hour reading of the first challenge. Due to the unequivocal findings observed after the first challenge, no second challenge was performed. Based on the study results, FAT 40400/A is not considered as skin sensitizer.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27 March 1990 - 13 June 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Data from a reliable in vivo test conducted before the enforcement of Commission Regulation (EU) 640/2012 of 06 July 2012 amending, for the purpose of its adaptation to technical progress, Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) are available.
Specific details on test material used for the study:
Identification Code: FAT 40400/A
Batch No.: BG 3247/TV6
Expiration date: 03/95
Storage Conditions: In the original container, at room temperature, protected from light
Stability of test article in vehicle: stable for at least 2 hours
Species:
guinea pig
Strain:
Himalayan
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wölferstrasse 4, CH-4414 Füllinsdorf.
- Age at acclimatization: Males: 7 weeks, Females: 8 weeks
- Weight at acclimatization: Males: 258-302 g, Females: 271-354 g
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: Pelleted standard 342 Kliba, Batch 57/90 guinea pig breeding/ maintenance diet ("Kliba", Klingentalmühle AG, CH-4303 Kaiseraugst, ad libitum.
- Water: Community tap water from Itingen ad libitum
- Acclimation period: One week under test conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
Three pairs of intradermal injections (0.1 ml/site) were made at the border of a 4 x 6 cm area.
5 % with physiological saline
Day(s)/duration:
Day 1
Adequacy of induction:
highest technically applicable concentration used
Route:
epicutaneous, semiocclusive
Vehicle:
physiological saline
Concentration / amount:
15 % in physiological saline
A 2 x 4 cm patch of filter paper was saturated with the test article
Day(s)/duration:
Day 8
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, semiocclusive
Vehicle:
physiological saline
Concentration / amount:
Two patches ( 2 x 2 cm) of filter paper were saturated with 10% in physiological saline of the test article and with the vehicle only and applied to the (a) left flank and (b) right flank
Day(s)/duration:
Day 22
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Ten animals (5 /sex) were treated with the vehicle alone and 20 animals (10/sex) were treated with the test article.
Details on study design:
RANGE FINDING TESTS:
- Intradermal injections: Intradermal injections (0.1 mL/site) were made into the clipped flank of two guinea-pigs at concentrations of 1, 3 and 5 % of the test article in physiological saline. The resulting dermal reactions were assessed 24 hours later.
- Epidermal applications: Patches of filter paper ( 2 x 2 cm) were saturated with concentrations of 5, 10, 15 and 25 % of the test article in physiological saline and applied to the clipped and shaved flanks of each of four guinea-pigs. The patches were covered by a strip of aluminum foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. The dressings were removed after an exposure period of 24 hours and the reaction sites were assessed for erythema and edema . Further examination of the sites were performed 24 and 48 hours after removal of the dressings.

MAIN STUDY
A. INDUCTION EXPOSURE
Intradermal injections: An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. Three pairs of intradermal injections (0.1 mL/site) were made at the border of a 4 x 6 cm area in the clipped region as follows: 1) Freunds' complete adjuvant 50:50 with bi-distilled water. 2) The test article, diluted to 5 % with physiological saline. 3) The test article at the concentration used in (2), emulsified in a 50:50 mixture of Freunds' complete adjuvant, and the vehicle used in (2).
Epidermal applications: One week after the injections, the scapular area (approximately 6 x 8 cm) was again clipped and shaved free of hair. A 2 x 4 cm patch of filter paper was saturated with the test article (15 % in physiological saline) and placed over the injection sites of the test animals. The patch was covered by aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The dressings were left in place for approximately 48 hours. Reaction sites were assessed for erythema and edema immediately, 24 and 48 hours after removal of the dressing.

B. CHALLENGE EXPOSURE
The test and control guinea-pigs were challenged two weeks after the epidermal induction application. Hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea-pig. Two patches ( 2 x 2 cm) of filter paper were saturated with a) non-irritant concentration (10 % in physiological saline of the test article) and b) with the vehicle alone and applied to the left (a) flank and right (b) flank using the same method as for the epidermal application. The dressings were removed approximately 24 hours later. The sites were assessed for erythema and edema immediately, 24 and 48 hours after removal.
Challenge controls:
The control animals were treated in the same way as described above without the test substance.
Positive control substance(s):
yes
Remarks:
A control group (Formaldehyde-solution) is tested twice a year for sensitivity check of the guinea pig strain. The most recent test was run at November/December, 1989 (RCC 253912).
Positive control results:
According to the results observed it is considered that the formaldehyde solution possess an extreme skin sensitizing (contact allergenic) potential in the guinea pig strain used (90 % positive).
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
Erythema
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
Erythema
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
15 %
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
other: Formaldehydloeung (HCHO)
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Remarks on result:
not measured/tested

Mortality: No death occurred during the study.


Local symptoms


- Vehicle control: Application area around the injection sites 1 and 3 was found to show erythema and edema from day 2 to 7; necroses were observed from day 8 to 11 and desiccation from day 11 to 18 and exfoliation from day 12 to 25 (termination of test). In addition staining was observed on first challenge application area from day 23 to 25.


- Test article: Application area around the injection site 1 was found to show erythema and edema from day 2 to 8; necroses days 8 to 11; desiccation days 11 to 18 and exfoliation from day 19 to 25 (termination of test). On injection site 2 erythema, edema and discoloration were observed from day 2 to 7; necroses from day 8 to 13; desiccation days 14 to 16 and exfoliation days 17 to 19. Injection site 3 was found to show erythema, edema and discoloration from day 2 to 7; necroses days 8 to 11; desiccation days 11 to 18 and exfoliation days 19 to 25. Further discoloration was observed on the epidermal induction or first challenge application area from days 11 to 18 respectively 23 to 25. On day 9 of test no observation could be performed because the animals were treated semi-occlusively.


Systemic symptoms were not observed. The body weight gain of the animals was not affected adversely during the study.

Interpretation of results:
GHS criteria not met
Conclusions:
FAT 40400/A is not considered as skin sensitizer.
Executive summary:

In a GLP-compliant sensitivity study using the Maximization-Test, performed according to OECD guideline 406 and EU method B.6, guinea pigs were treated with FAT 40400/A. Twenty animals were treated with the test substance and ten animals with only the vehicle (physiological saline). The concentrations of the test substance used in the main study were determined by the results of the preliminary study. The intradermal induction of sensitization in the test group was performed using 5 % of the test substance in both the vehicle and adjuvant/vehicle mixture. One week later this was boosted by the topical application of the test substance at 15 % concentration over the injection sites. Animals of the control group were treated in the same manner but only the selected vehicle was used. Two weeks after the second induction all animals were challenged by topical application of the test substance at 10 % concentration. No death occurred during the study. Application area around the injection site 1of test article treated animal was found to show erythema and edema from day 2 to 8; necroses days 8 to 11; desiccation days 11 to 18 and exfoliation from day 19 to 25 (termination of test). On injection site 2 erythema, edema and discoloration were observed from day 2 to 7; necroses from day 8 to 13; desiccation days 14 to 16 and exfoliation days 17 to 19. Injection site 3 was found to show erythema, edema and discoloration from day 2 to 7; necroses days 8 to 11; desiccation days 11 to 18 and exfoliation days 19 to 25. Further discoloration was observed on the epidermal induction or first challenge application area from days 11 to 18 respectively 23 to 25. On day 9 of test no observation could be performed because the animals were treated semi-occlusively. Systemic symptoms were not observed. The body weight gain of the animals was not affected adversely during the study. Two out of 20 animals showed a positive erythema reaction at the 24 and 48 hour reading of the first challenge. Due to the unequivocal findings observed after the first challenge, no second challenge was performed. Based on the study results, FAT 40400/A is not considered as skin sensitizer.

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

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the findings in the skin sensitisation study, the test substance does not need to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.