TROYSOL LAC

Endpoint summary

Description of key information

Based on the findings of the Dermal Sensitization test (Buehler method) the test item is not considered to be a skin sensitizer.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

December 20, 1994 - January 27, 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 81-6 (Skin Sensitisation)

Version / remarks:

Pesticide Assessment Guidelines, Subdivision F: Hazard Evaluation: Human and Domestic Animals, Section 81-6 OECD Guideline 406, Buehler Test

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

The Buehler Test (1995) met the previous requirements before the entry into force of REACH. The Buehler Test is suitable and reliable to cover this endpoint. For this reason and for animal welfare reasons, no further in vivo study (LLNA test) needs to be performed.

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Davidson's Mill Farms, South Brunswick, NJ
- Weight at study initiation: 335 – 388 g (males), 288 - 413 g (females)
- Housing: The animals were group housed in suspended stainless steel caging with mesh floors.
- Diet: Pelleted Purina Guinea Pig Chow
- Water: ad libitum (tap water)
- Acclimation period: 6 or 7 days
- Indication of any skin lesions: Only healthy animals without pre-existing skin irritation were selected for test.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 – 22.2
- Photoperiod (hrs dark / hrs light): 12/12

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

50% and 75% (0.4 mL)

Day(s)/duration:

Two times each week (6 hours) for three weeks

Adequacy of induction:

other: Due to the severity of irritation noted at all test sites, the concentration was reduced to 50% after the second induction phase .

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

20%

Day(s)/duration:

Thirteen days after last induction/ 6 hours

Adequacy of challenge:

highest non-irritant concentration

No.:

#2

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

20%

Day(s)/duration:

Seven days after challenge #1

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:

RANGE FINDING TESTS: The fur was removed by clipping the dorsal area and flanks of each guinea pig. This area was divided into four test sites (two sites on each side of the midline) on each animal. The test substance was applied neat (100%) and was also diluted with distilled water to yield concentrations of 75%, 50%, 25%, 20%,10%, 5% and 2%. Each concentration was applied to a test site using an occlusive 25 mm Hilltop Chamber. The sites were wrapped with non-allergenic Durapore adhesive tape. After 6 hours of exposure, the chambers were removed. Twenty-four hours after application, each site was evaluated for local reactions (erythema). Based on these results, the highest non-irritating concentration selected for the challenge phase was a 20% w/w suspension in distilled water. Furthermore, the selected highest non-irritating concentration of the positive control (1-Chloro-2,4-Dinitrobenzene (DNCB)) was determined to be a 0.04% w/w solution in acetone.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 6
- Exposure period: 6 hours
- Test groups: treatment with the test item
- Site: left side of each animal
- Frequency of applications: two times per week
- Duration: three weeks
- Concentrations: 75% test item in water (concentration was reduced to 50% after the second induction phase)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: thirteen days after last induction
- Exposure period: 6 hours
- Test groups: treatment with test item
- Control group: ten animals were treated with 20% test item suspension
- Site: right site of each animal
- Concentrations: 20% test item in water
- Evaluation (hr after challenge): 24 and 48

C. RECHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: seven days after challenge
- Exposure period: 6 hours
- Test groups: treatment with test item
- Control group: An additional group of ten animals was placed on test to serve as a naive group for rechallenge and treated with the test item suspension.
- Site: naive right site of each animal
- Concentrations: 20% test item in water
- Evaluation (hr after challenge): 24 and 48

Challenge controls:

An additional group of ten animals was placed on test to serve as a naive group for rechallenge.

Positive control substance(s):

yes

Remarks:

1-Chloro-2,4-Dinitrobenzene

Positive control results:

INDUCTION PHASE
Positive Control Animals (0.08% DNCB in 80% aqueous ethanol): Very faint to severe erythema (0.5-3) was exhibited at all positive control sites during the induction phase. Eschar and/or desquamation were evident at several sites after the third and fifth inductions. Due to the severity of irritation, the dose sites were relocated to an adjacent area for inductions four and six.

CHALLENGE PHASE
Positive Control Animals (0.04% DNCB in acetone): Seven of twenty positive control animals exhibited signs of a sensitization response (faint to moderate erythema (1-2)) 24 hours after challenge. Similar indications persisted in three of these animals through 48 hours. Very faint erythema (0.51) was noted at most other positive control sites following the challenge dose.

Positive Naive Control Animals (0.04% DNCB in acetone); Very faint erythema (0.5) was noted at Three of ten positive naive control animals 24 hours after challenge. Irritation cleared from all affected sites by 48 hours.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

20 %

No. with + reactions:

1

Total no. in group:

20

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

20

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

test chemical

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

20

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

20

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

negative control

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

negative control

Dose level:

20 %

No. with + reactions:

0

Total no. in group:

10

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the findings of the Dermal Sensitization test (Buehler method) the test item is not considered to be a skin sensitizer.

Executive summary:

A Dermal Sensitization test was conducted with guinea pigs to determine the potential for the test substance to produce sensitization after repeated topical applications.

The test substance was applied to twenty healthy test guinea pigs and Dinitrochlorobenzene (DNCB, 0.08%), respectively, was applied to twenty healthy positive control animals two times each week for a three week induction period. Thirteen days after the last induction, a challenge dose of the test and positive control substances at their highest non-irritating concentrations (HNIC, 20% and 0.04%, respectively) was applied to a naive site on each guinea pig. Approximately

24 and 48 hours after each induction and challenge dose, the animals were scored for erythema.

Two naive control groups (ten animals for the test substance and ten for the positive control substance) were maintained under the same environmental conditions and treated with the test or positive control substance at challenge only.

Very faint to severe erythema (score 0.5-3), was noted at all test sites during the induction phase. Irritation was most severe following the second, third and fifth inductions. Eschar and/or desquamation were also evident at several sites following these doses. A reduction in the severity of irritation was noted shortly after the induction concentration was reduced (75% to 50% suspension) or after the dose sites were relocated to an adjacent area.

Very faint 10 severe erytheme (score 0.5-3) was exhibited at all positive control sites during the induction phase. Eschar and/or desquamation were evident at some sites after the third and fifth inductions. Due to the severity of irritation, the dose sites were relocated to an adjacent area for inductions four and six.

Faint erythema (score 1) was noted at one test site 24 hours after the challenge dose. Very faint erythema (score 0.5) was noted at most other sites 24 and 48 hours after challenge.

Very faint erythema (score 0.5) was noted at several naive control lest sites alter the challenge

dose.

Seven of twenty positive control animals exhibited signs of a sensitization response (faint to moderate erythema (score 1-2)) 24 hours after challenge. Similar indications persisted in three of these animals through 48 hours. Very faint erythema (score 0.5) was noted at most other positive control sites following the challenge dose. Very faint erythema (score 0.5) was noted at three of ten positive naive control animals 24 hours after challenge. Irritation cleared from all affected sites by 48 hours,

Due to the presence of taint erythema (score 1) at one test site 24 hours after challenge, a rechallenge was conducted. An additional group of ten animals was placed on test to serve as a naive control group for rechallenge.

Very faint (score 0.5) erythema was noted at five of twenty test sites after rechallenge. Erythema cleared from two of the five affected sites by 48 hours.

 

Very faint erythema (score 0.5) was noted at three naive control sites 24 and 48 hours after rechallenge.

Based on the results of testing, the test substance is not considered to be a contact sensitizer.

 

Reference 2

Endpoint:

skin sensitisation: in vitro

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

A Dermal Sensitization test was conducted with guinea pigs to determine the potential for the test substance to produce sensitization after repeated topical applications.

The test substance was applied to twenty healthy test guinea pigs and Dinitrochlorobenzene (DNCB, 0.08%), respectively, was applied to twenty healthy positive control animals two times each week for a three week induction period. Thirteen days after the last induction, a challenge dose of the test and positive control substances at their highest non-irritating concentrations (HNIC, 20% and 0.04%, respectively) was applied to a naive site on each guinea pig. Approximately

24 and 48 hours after each induction and challenge dose, the animals were scored for erythema.

Two naive control groups (ten animals for the test substance and ten for the positive control substance) were maintained under the same environmental conditions and treated with the test or positive control substance at challenge only.

Very faint to severe erythema (score 0.5-3), was noted at all test sites during the induction phase. Irritation was most severe following the second, third and fifth inductions. Eschar and/or desquamation were also evident at several sites following these doses. A reduction in the severity of irritation was noted shortly after the induction concentration was reduced (75% to 50% suspension) or after the dose sites were relocated to an adjacent area.

Very faint 10 severe erytheme (score 0.5-3) was exhibited at all positive control sites during the induction phase. Eschar and/or desquamation were evident at some sites after the third and fifth inductions. Due to the severity of irritation, the dose sites were relocated to an adjacent area for inductions four and six.

Faint erythema (score 1) was noted at one test site 24 hours after the challenge dose. Very faint erythema (score 0.5) was noted at most other sites 24 and 48 hours after challenge.

Very faint erythema (score 0.5) was noted at several naive control lest sites alter the challenge

dose.

Seven of twenty positive control animals exhibited signs of a sensitization response (faint to moderate erythema (score 1-2)) 24 hours after challenge. Similar indications persisted in three of these animals through 48 hours. Very faint erythema (score 0.5) was noted at most other positive control sites following the challenge dose. Very faint erythema (score 0.5) was noted at three of ten positive naive control animals 24 hours after challenge. Irritation cleared from all affected sites by 48 hours,

Due to the presence of taint erythema (score 1) at one test site 24 hours after challenge, a rechallenge was conducted. An additional group of ten animals was placed on test to serve as a naive control group for rechallenge.

Very faint (score 0.5) erythema was noted at five of twenty test sites after rechallenge. Erythema cleared from two of the five affected sites by 48 hours.

 

Very faint erythema (score 0.5) was noted at three naive control sites 24 and 48 hours after rechallenge.

Based on the results of testing, the test substance is not considered to be a contact sensitizer.

 

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

Based on the available experimental data, the test item is not considered to be classified for skin sensitization under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation (EC) No 2016/1179.