Registration Dossier

Administrative data

Description of key information

The source substance is not sensitising to the skin.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1987
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
None
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Principles of method if other than guideline:
None
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
None
Specific details on test material used for the study:
FAT 40279
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG CH 4414 Fuellinsdorf / Switzerland
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: Males: 375 - 473 g Females: 376 - 495 g
- Housing: Plakrolon type-3 cages with standard softwood bedding
- Diet (e.g. ad libitum): guinea pig diet ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum.
- Acclimation period: One week under test conditions after veterinary examination


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-3 °C
- Humidity (%): 40-70 %
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light):12 hours artificial fluorescent light/12 hours dark, music/light period.
Route:
intradermal
Vehicle:
physiological saline
Remarks:
5%
Concentration / amount:
emulsified in a 50:50 mixture of Freunds' complete adjuvant,
Day(s)/duration:
Test Day 01
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
other: topical
Vehicle:
physiological saline
Concentration / amount:
25%
Day(s)/duration:
Test Day 08
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
25%
Day(s)/duration:
Test Day 22
Adequacy of challenge:
highest non-irritant concentration
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
25%
Day(s)/duration:
Test Day 36
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 animals per dose
Details on study design:
None
Challenge controls:
None
Positive control substance(s):
yes
Remarks:
DNCB
Positive control results:
None
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25%
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:
48
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation

None

Interpretation of results:
GHS criteria not met
Conclusions:
FAT 40279/B is found to be non sensitiser.
Executive summary:

A supporting study was performed to assess the allergenic potential of FAT 40279/B in albino guinea pigs. 5 males, 5 females for the vehicle control group and 10 males, 10 females for the test article group. A control group is tested twice a year as a sensitivity check of the guinea pig strain. The last test was run during February 1987. The challenge site was evaluated just after application, 24 and 48 hours after removal of the patch. The readings were made under artificial fluorescent light (daylight spectrum). Redness constitutes the minimum criterion of an allergic reaction. Strongly sensitized animals displayed a vivid redness, associated with indurated swelling. The reactions were scored on the basis of the Draize score described under preliminary study. No positive reactions mere observed after the first and second challenge application. Therefore, FAT 40279 is considered to be non sensitiser.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
None
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
None
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
None
Specific details on test material used for the study:
None
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 study initiation: males : 7 weeks females: 8 weeks
- Weight at study initiation: males: 311 - 393 g females: 302 - 402 g
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel",
Schill AG, CH-4132 Muttenz).
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period: One week under test conditions after veterinary examination.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-3°C
- Humidity (%): 40-70 %,
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
Test group:
1) Freund's complete adjuvant 50:50 with bi-distilied 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 Freund's complete adjuvant and the vehicle used in (2).

Control Group:
1) Freund's complete adjuvant 50:50 with bi-distilled water.
2) Vehicle used in (2) for test group.
3) Freund's complete adjuvant 50:50 with bi-distilled water.
Day(s)/duration:
Test Day 01
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
25% in petrolatum-oil
Day(s)/duration:
Test Day 08
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
25 % in petrolatum-oil
Day(s)/duration:
Test Day 22
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 animals per dose
Details on study design:
None
Challenge controls:
None
Positive control substance(s):
yes
Remarks:
Solution of formaldehyde
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25% IN PETROLATUM-OIL
No. with + reactions:
1
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
1
Total no. in group:
20

None

Interpretation of results:
GHS criteria not met
Conclusions:
FAT 40000/F is found to be non sensitiser.
Executive summary:

A supporting study was performed to assess the allergenic potential of FAT 40000/F in albino guinea pigs. The Maximization-Test protocol of B. Magnusson and A.M. Kligman (1970) was used. Ten animals (5 males, 5 females) were used as control group and 20 animals (10 males, 10 females) were used as test group. Prior to the first reading of the challenge reactions, the skin was gently washed-off with petrolatum-oil to clean the application site from staining produced by the test article, so that possible erythema reactions were clearly visible at that time. Due to the unequivocal findings observed after the first challenge, no second challenge was performed. No toxic symptoms were evident in the guinea pigs of the control or test group. No death occurred. From the results described above no allergenic potency of the test article FAT 40000/F was concluded. The results were interpreted according to the rating of Magnusson and Kligman (1970). According to CLP classification criteria, this test article is considered to be not a sensitizer.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
None
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
None
GLP compliance:
no
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
None
Specific details on test material used for the study:
None
Species:
guinea pig
Strain:
Himalayan
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, CH-4414 Füllinsdorf
- Age at study initiation: 6 - 7 weeks
- Weight at study initiation: Control and test group 376 - 449 g Pretest 410 - 437 g
- Housing: Individually in Makrolon type-3 cages (size: 27 x 42 x 15 cm) with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Kliba 342, Batch 70/92 guinea pig breeding/ maintenance diet ("Kliba", Klingentalmühle AG, CH-4303 Kaiseraugst), ad libitum. Results of
analysis for contaminants are included in this report.
- Water (e.g. ad libitum): Community tap water from Füliinsdorf, ad libitum. Once weekly additional supply of ascorbic acid (1 g/1) via the drinking water. Results of bacteriological,
chemical and contaminant analyses are included in this report.
- Acclimation period: One week for the control and test group under test conditions after veterinary examination. Two days under test conditions after veterinary examination for the pretest animals. Only animals without any visual signs of illness were used for the study.
- Indication of any skin lesions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-3°C
- Humidity (%): 40-70 %,
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light):12 hours artificial fluorescent light (approx. 100 lux) /12
hours dark, music during the light period
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
Test group:
1) Freund's complete adjuvant 50:50 with physiological saline.
2) The test article, diluted to 5% with physiological saline.
3) The test article diluted to 5% by emulsion in a 50:50 mixture of Freund's complete adjuvant and physiological saline.

Control Group:
1) Freund's complete adjuvant 50:50 with physiological saline.
2) Physiological saline.
3) Freund's complete adjuvant 50:50 with physiological saline.
Day(s)/duration:
Test Day 01
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: Vaselinum album
Concentration / amount:
25% in vaselinum album
Day(s)/duration:
Test Day 07
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: vaselinum album
Concentration / amount:
25% in vaselinum album
Day(s)/duration:
Test Day 22
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 animals per dose
Details on study design:
None
Challenge controls:
None
Positive control substance(s):
yes
Remarks:
2-MERCAPTOBENZOTHIAZOL
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25% IN VASELINUM ALBUM
No. with + reactions:
10
Total no. in group:
19
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25% IN VASELINUM ALBUM
No. with + reactions:
8
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation

None

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
FAT 40000/G is found to be a skin sensitiser.
Executive summary:

A supporting study was performed to determine the skin sensitisation of FAT 40000/G in rabbits according to OECD Guideline 406. From the results described above moderate allergenic potency of the test article FAT 40000/G was concluded. The results were interpreted acccrding to the rating of Magnusson and Kligman (1969). The response of at least 30 % positive animals is considered positive "R43" using the described Maximization-test method following the "Legislation on Dangerous Substances, Classification and Labelling in the European Communities", Directive 67/548/EEC. In this study 53% of the animals were positive after the first challenge at a test substance concentration of 25 %. According to EEC (European Economic Community) classification criteria described in guidelines 83/467, June 29, 1983 (Official Journal L 257 of September 16, 1983) and 67/548, June 27, 1967 (Official Journal 196 of August 16, 1967), this test article is considered to be a sensitizer.

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
None
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
22 July 2010
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
2003
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147, amended 10 December 2002
Version / remarks:
10 December 2002
Deviations:
no
Principles of method if other than guideline:
None
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Approximately -20”C in the dark
Species:
mouse
Strain:
CBA/Ca
Remarks:
CBA/CaOlaHsd
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS B.V., Inc., Horst, The Netherlands
- Females (if applicable) nulliparous and non-pregnant: yes

- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15-23 g
- Housing: suspended solid floor polypropylene cages furnished with softwood woodflakes
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: >5 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25”C
- Humidity (%): 30-70%
- Air changes (per hr): >15
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES: From: 06 June 2016 To: 19 July 2016
Vehicle:
other: 1% pluronic L92 in distilled water
Concentration:
Preliminary Screening Test = 25% w/w in 1% pluronic L92 in distilled water

Main Test = 25%, 10% or 5% w/w in 1% pluronic L92 in distilled water
No. of animals per dose:
Preliminary Screening Test = 1 mouse

Main Test = 4 mice per group
Details on study design:
Preliminary Screening Test
As no toxicological information was available regarding the systemic toxicity/irritancy potential of the test item, a preliminary screening test was performed using one mouse. The mouse was treated by daily application of 25 µL of the test item at a concentration of 25% w/w in 1% pluronic L92 in distilled water, to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). The mouse was observed twice daily on Days 1, 2 and 3 and once daily on Days 4, 5 and 6. Local skin irritation was scored daily according to the scale included below. Any clinical signs of toxicity, if present, were also recorded. The body weight was recorded on Day 1 (prior to dosing) and on Day 6.
The thickness of each ear was measured using a Mitutoyo 547 300S gauge (Mitutoyo Corporation), pre dose on Day 1, post dose on Day 3 and on Day 6. Any changes in the ear thickness were noted. Mean ear thickness changes were calculated between time periods Days 1 and 3 and Days 1 and 6. A mean ear thickness increase of equal to or greater than 25% was considered to indicate excessive irritation and limited biological relevance to the endpoint of sensitization.

Main Test
Test Item Administration
Groups of four mice were treated with the test item at concentrations of 25%, 10% or 5% w/w in 1% pluronic L92 in distilled water. The preliminary screening test suggested that the test item would not produce systemic toxicity or excessive local skin irritation at the highest suitable concentration. The mice were treated by daily application of 25 µL of the appropriate concentration of the test item to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). The test item formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.
A further group of four mice received the vehicle alone in the same manner.

3H-Methyl Thymidine Administration
Five days following the first topical application of the test item or vehicle (Day 6) all mice were injected via the tail vein with 250 µL of phosphate buffered saline (PBS) containing 3H methyl thymidine (3HTdR: 80 µCi/mL, specific activity 2.0 Ci/mmoL, ARC UK Ltd) giving a total of 20 µCi to each mouse.

Observations
Clinical Observations: All animals were observed twice daily on Days 1, 2 and 3 and on a daily basis on Days 4, 5 and 6. Any signs of toxicity or signs of ill health during the test were recorded.
Body Weights: The body weight of each mouse was recorded on Day 1 (prior to dosing) and Day 6 (prior to termination).

Terminal Procedures
Termination: Five hours following the administration of 3HTdR all mice were killed by carbon dioxide asphyxiation followed by cervical separation. The draining auricular lymph nodes from the four mice were excised and pooled for each experimental group. For each group 1 mL of PBS was added to the pooled lymph nodes.

Preparation of Single Cell Suspension: A single cell suspension of pooled lymph node cells was prepared by gentle mechanical disaggregation through a 200 mesh stainless steel gauze. The lymph node cells were rinsed through the gauze with 4 mL of PBS into a petri dish labeled with the study number and dose concentration. The lymph node cell suspension was transferred to a centrifuge tube. The petri dish was washed with an additional 5 mL of PBS to remove all remaining lymph node cells and these were added to the centrifuge tube. The pooled lymph node cells were pelleted at 1400 rpm (approximately 190 g) for 10 minutes.
The pellet was re suspended in 10 mL of PBS and re pelleted. To precipitate out the radioactive material, the pellet was re suspended in 3 mL of 5% Trichloroacetic acid (TCA).

Determination of 3HTdR Incorporation:
After approximately 18 hours incubation at approximately 4 C, the precipitates were recovered by centrifugation at 2100 rpm (approximately 450 g) for 10 minutes, re suspended in 1 mL of TCA and transferred to 10 mL of scintillation fluid. 3HTdR incorporation was measured by scintillation counting. The "Poly Q™" vials containing the samples and scintillation fluid were placed in the sample changer of the scintillator and left to stand in darkness for approximately 20 minutes. The purpose of this period of time in darkness was to reduce the risk of luminescence, which has been shown to affect the reliability of the results. After approximately 20 minutes, the vials were shaken vigorously. The number of radioactive disintegrations per minute was then measured using the Beckman LS6500 scintillation system (Beckman Instruments Inc, Fullerton, CA, USA).

Data Evaluation
The proliferation response of lymph node cells was expressed as the number of radioactive disintegrations per minute per lymph node (disintegrations per minute/node) and as the ratio of 3HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes (Stimulation Index).
The test item will be regarded as a sensitizer if at least one concentration of the test item results in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test item failing to produce a threefold or greater increase in 3HTdR incorporation will be classified as a "non sensitizer".
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
None
Positive control results:
α Hexylcinnamaldehyde, tech., 85% was considered to be a sensitizer under the conditions of the test with a Stimulation Index of 4.47
Key result
Parameter:
SI
Value:
1.25
Test group / Remarks:
5% w/w
Remarks on result:
other: Negative
Key result
Parameter:
SI
Value:
1.65
Test group / Remarks:
10% w/w
Remarks on result:
other: Negative
Key result
Parameter:
SI
Value:
2.33
Test group / Remarks:
25% w/w
Remarks on result:
other: Negative
Cellular proliferation data / Observations:
Preliminary Screening Test
No signs of systemic toxicity, visual local skin irritation or irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted.
Based on this information the dose levels selected for the main test were 25%, 10% and 5% w/w in 1% pluronic L92 in distilled water.

Main Test
Estimation of the Proliferative Response of Lymph Node Cells
The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

Concentration (%w/w) in
1% pluronic L92 in distilled water Stimulation Index Result
5 1.25 Negative
10 1.65 Negative
25 2.33 Negative

There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test.

Body weight change of the test animals between Day 1 and Day 6 was comparable to that observed in the corresponding control group animals over the same period.

None

Interpretation of results:
GHS criteria not met
Conclusions:
The test item was considered to be a non-sensitizer under the conditions of the test.
Executive summary:

A study was performed to assess the skin sensitization potential of the test item in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear.

Methods

Following a preliminary screening test in which no clinical signs of toxicity were noted at a concentration of 25% w/w, this concentration was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of four animals, were treated with 50 µL (25 µL per ear) of the test item as a solution in 1% pluronic L92 in distilled water at concentrations of 25%, 10% or 5% w/w. A further group of four animals was treated with 1% pluronic L92 in distilled water alone.

Results

The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

 Concentration (%w/w) in 1% pluronic L92 in distilled water  Stimulation Index  Result
 5  1.25  Negative
 10  1.65  Negative
 25  2.33  Negative

Conclusion

The test item was considered to be a non-sensitizer under the conditions of the test. The test item does not meet the criteria for classification according to the Globally Harmonized Classification System.

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that the source and the target substance have very similar physicochemical and (eco)toxicological properties because their chemical structures are nearly identical. An analogue approach has thus been employed. The target substance is the meta-isomer of the dye Reactive Yellow 095, where the sulphonate group is bound at the meta-position of the amino benzene moiety. The source chemical is the reaction mass of both the meta-isomer and the para-isomer of Reactive Yellow 095.
The presence of sulphonate groups make both dyes highly water soluble and therefore less critical for human health and environmental issues. Based on their chemical similarity, similar properties are expected in both humans and the environment.
2. SOURCE AND TARGET CHEMICAL(S)
Source: Reactive Yellow 095 meta/para (CAS# --- / EC# 944-218-2)
Target: Reactive Yellow 095 meta (CAS# 84045-63-6 / EC# 281-865-3)
3. ANALOGUE APPROACH JUSTIFICATION
see attachment under 4.12 Auto flammability
Reason / purpose:
read-across source
Parameter:
SI
Value:
1.25
Test group / Remarks:
5% (w/w)
Remarks on result:
other: negative
Parameter:
SI
Value:
1.65
Test group / Remarks:
10% (w/w)
Remarks on result:
other: negative
Parameter:
SI
Value:
2.33
Test group / Remarks:
25% (w/w)
Remarks on result:
other: negative
Interpretation of results:
GHS criteria not met
Conclusions:
The substance was considered to be not sensitising under the conditions of the in vivo LLNA test.
Executive summary:

A study was performed to assess the skin sensitization potential of the source substance in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear.

Following a preliminary screening test in which no clinical signs of toxicity were noted at a concentration of 25% (w/w), this concentration was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of four animals, were treated with 50 µL (25 µL per ear) of the test item as a solution in 1% pluronic L92 in distilled water at concentrations of 25%, 10% or 5% (w/w). A further group of four animals was treated with 1% pluronic L92 in distilled water alone.

The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

 Concentration (%w/w) in 1% pluronic L92 in distilled water  Stimulation Index  Result
 5  1.25  Negative
 10  1.65  Negative
 25  2.33  Negative

The test item was considered to be a non-sensitizer under the conditions of the test. The test item does not meet the criteria for classification according to CLP.

The structurally related target substance will show the same behaviour and therefore it is anticipated that it does not need to be classified as a skin sensitiser either.

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

In total four studies have been conducted to investigate skin sensitization potential of Reactive Yellow 095.

Three tests followed the testing protocol of the Guinea Pig Maximisation Test according to Magnusson & Kligman: batch FAT 40000/F was tested in 1990 and batch FAT 40000/G was tested in 1992, both batches consisting of equivalent amounts of the meta- and para-isomer of Reactive Yellow 95. Batch FAT 40279/B, which is the pure para compound of Reactive Yellow 95, was tested in 1987. The fourth and most recent test was conducted in 2016 on batch FAT 40000/I where mice were tested following the protocol of the Local Lymph Node Assay (LLNA). Batch FAT 40000/I consists of equivalent amounts of the meta- and para-isomer of the dye.

 

While the skin sensitization test according to the protocol of the LLNA gave a negative result in mice, the results of the GPMT studies of batch 40000/G and 40000/F gave contradictory results. The first test with batch 40000/F has been judged negative with no animals being sensitized in the test groups, while the second test with batch 40000/G showed a positive result with 10 out of 19 animals being judged "positive" after the first challenge at a test substance concentration of 25%. In addition one animal of the test group died for unknown reasons on day 5 of the study. After necropsy no evidence was found that the death was substance related, therefore this result was excluded from the final evaluation and rated as incidental.

The following differences were observed between the two studies:

During the intradermal pretest in 2 animals, the substance was tested at 5%, 3% and 1% in both studies. FAT 40000/F did not induce any erythema nor edema. FAT 40000/G did induce very slight edema, while erythema was not scored due to yellow staining.

During the epidermal pretest performed with 4 animals, the substance was tested at 25, 15, 10 and 5% in both studies. No erythema nor edema was noted in all animals for all four concentrations in both studies. Therefore, the 25% concentration was selected as highest non-irritant concentration to be used for the challenge phase but was also used as highest applicable concentration to be used for the induction phase.

In the main study of FAT 40000/F, after being treated with 10% SLS, slight erythema was shown in some of the control animals and treated animals after epidermal induction. In challenged animals, only 1 FAT 40000/F-treated animal showed slight erythema. After induction in the FAT 40000/G study, only 6 test substance treated animals showed very slight oedema, while erythema was not scored due to yellow staining. After the challenge, only in FAT 40000/G-treated animals, 10 out of 19 animals showed slight erythema, while only 1 animal showed very slight edema. It is strange that the skin was not depilated after epidermal induction with FAT 40000/G in order to score erythema as well. It is clear that more animals showed edema after epidermal induction than after the epidermal challenge with FAT 40000/G. This may indicate a false positive result.

 

The 1987 Guinea Pig Maximization Test with FAT 40279/B (representing the pure para-form of the molecule) also gave a negative response with no single animal showing signs of sensitziation, signs of toxicity or death in response to the challenge procedure.

Taking into account that the LLNA method is generally preferred over the GPMT method, especially for coloured substances, we decided to use the existing LLNA study as the key study. The LLNA study showed that Reactive Yellow 95 is not sensitising to the skin, while two of the three GPMT studies showed the same.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The source substance is judged to be not sensitising to the skin, based on a LLNA key study and two supporting GPMT studies. It is anticipated that the structurally related target substance will show the same behaviour. Therefore it will not be classified for skin sensitisation either.