Di-tert-butyl trisulphide

Endpoint summary

Description of key information

The registered substance was moderately irritating following a single dose with 4-hour semi-occluded exposure in rabbits (OECD TG 404) and very slightly irritating to rabbit eyes 24 hours after dosing (OECD TG 405).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole du Val de Selle (80160 Prouzel, France)
- Age at study initiation: no data
- Weight at study initiation: 2.5 +/- 0.1 kg
- Housing: individually in polystyrene cages
- Diet (e.g. ad libitum): Rabbits sustenance 112 C (UAR, 91360 Villemoisson/Orge, France)
- Water (e.g. ad libitum): filtered tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 50 +/- 20
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 ml

Duration of treatment / exposure:

4 hour(s)

Observation period:

up to 15 days

Number of animals:

3

Details on study design:

One male New Zealand White rabbit received on a 6 cm2 clipped area, a single dose of 0.5 ml of the test substance in its original form and prepared on a dry compress. The test substance was held in contact with the skin for 4 hours by means of a semi-occlusive dressing. The cutaneous reactions were observed one hour, 24, 48 and 72 hours after removal of the dressing and then daily in order to observe their reversibility or irreversibility. No residual test substance was observed after removal of the dressing. As the test substance was not very irritant, the first results were confirmed on 2 other animals. The mean value of the scores recorded after 24, 48 and 72 hours for each animal was calculated.

Irritation parameter:

edema score

Basis:

animal: #1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Very slight and well-defined erythema were observed after application of the test substance and reversed on day 7. No oedema was noted. The mean score (24+48+72 h) for erythema was 0.3 in one animal and 2.0 in 2 animals.

Other effects:

A dryness of the skin was observed between days 4 and 15.

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

TPS 44 was considered as a mild irritant when administered by cutaneous route in rabbits.

Executive summary:

The cutaneous irritation that could be induced by ditert-butyl polysulfides (TPS 44) was evaluated in rabbits according to the O.E.C.D. Guideline No. 404 for the testing of chemicals and GLP. One male New Zealand White rabbit received on a 6 cm2 clipped area, a single dose of 0.5 ml of TPS 44 in its original form and prepared on a dry compress. The test substance was held in contact with the skin for 4 hours by means of a semi-occlusive dressing. The cutaneous reactions were observed one hour, 24, 48 and 72 hours after removal of the dressing and then daily in order to observe their reversibility or irreversibility. No residual test substance was observed after removal of the dressing. As the test substance was not very irritant, the first results were confirmed on 2 other animals. The mean value of the scores recorded after 24, 48 and 72 hours for each animal was calculated.

Very slight and well-defined erythema were observed after application of TPS 44 and reversed on day 7. No oedema was noted. The mean score for erythema was 0.3 in one animal and 2.0 in 2 animals. A dryness of the skin was observed between days 4 and 15. TPS 44 was considered as a mild irritant when administered by cutaneous route in rabbits.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Grimaud frères selection S.A.S., La Corbière, Roussay, Franc
- Age at study initiation: 2 to 4 months old
- Weight at study initiation: 2.4 ± 0.1 kg
- Housing: individually in polystyrene cages
- Diet (e.g. ad libitum): 110C pelleted diet (SAFE, Villemoisson, Epinay sur-Orge, France)
- Water (e.g. ad libitum): Filtered tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3
- Humidity (%): 30 to 70
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12 h/12 h

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 ml

Duration of treatment / exposure:

3 minutes, 1 hour and 4 hours.

Observation period:

The skin was examined approximately 1 hour, 24, 48 and 72 hours after removal of the dressing.
Since there were persistent irritation reactions at 72 hours, the observation period was extended. In the absence of complete reversibility of skin reactions on day 15, the study was ended.

Number of animals:

3

Details on study design:

TPS 44 was first evaluated on a single animal (No. 382). The durations of exposure were 3 minutes, 1 hour and 4 hours.
Since the test item was not severely irritant on this first animal, it was then applied for 4 hours to two other animals (Nos. 751 and 752).

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 15 days

Remarks on result:

other: 4-hour exposure

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: 4-h exposure

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: 4-hour exposure

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: 4-h exposure

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 4-h exposure

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 8 days

Remarks on result:

other: 4-h exposure

Irritant / corrosive response data:

After a 3-minute exposure (one animal)
A very slight or well-defined erythema (grade 1 or 2) was noted from day 1 up to the end of the observation period (day 15).
A very slight or slight edema (grade 1 or 2) was observed from day 3 up to day 9.
Dryness of the skin was also recorded from day 5 up to the end of the observation period (day 15).

After a 1-hour exposure (one animal)
A very slight or well-defined erythema (grade 1 or 2) was noted from day 1 up to the end of the observation period (day 15).
A very slight or slight edema (grade 1 or 2) was observed from day 4 up to day 9.
Dryness of the skin was also recorded from day 6 up to the end of the observation period (day 15).

After a 4-hour exposure (three animals)
A very slight or well-defined erythema (grade 1 or 2) was noted in all animals from day 1 up to day 12, 13 or 15 (end of the observation period).
A very slight or slight edema (grade 1 or 2) was observed in 2/3 animals from day 1 up to day 3 or 7.
Dryness of the skin was also recorded in all animals between days 5 and 15 (end of the observation period).

Mean scores over 24, 48 and 72 hours for each animal were 2.0, 2.0 and 2.0 for erythema and 0.0, 1.0 and 2.0 for edema.

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

TPS 44 was a mild irritant when applied topically to rabbits.

Executive summary:

The potential of di-tert-butyl polisulfides (TPS 44) to induce skin irritation was evaluated in rabbits according to OECD No. 404 and EC (2004/73/EC, B.4) guidelines and in compliance with the principles of GLP. TPS 44 was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit. Since the test item was not severely irritant on this first animal, it was then applied for 4 hours to two other animals. A single dose of 0.5 mL of undiluted TPS 44 was applied to the closely-clipped skin of one flank. TPS 44 was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the end of the observation period. The mean values of the scores for erythema and edema were calculated for each animal.

After a 3-minute exposure (one animal), a very slight or well-defined erythema was noted from day 1 up to the end of the observation period (day 15). A very slight or slight edema was observed from day 3 up to day 9 and dryness of the skin was also recorded from day 5 up to the end of the observation period (day 15). After a 1-hour exposure (one animal), a very slight or well-defined erythema was noted from day 1 up to the end of the observation period (day 15). A very slight or slight edema was observed from day 4 up to day 9 and dryness of the skin was also recorded from day 6 up to the end of the observation period (day 15). After a 4-hour exposure (three animals), a very slight or well-defined erythema was noted in all animals from day 1 up to day 12, 13 or 15 (end of the observation period). A very slight or slight edema was observed in 2/3 animals from day 1 up to day 3 or 7 and dryness of the skin was also recorded in all animals between days 5 and 15 (end of the observation period). Mean scores over 24, 48 and 72 hours for each animal were 2.0, 2.0 and 2.0 for erythema and 0.0, 1.0 and 2.0 for edema.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole du Val de Selle (80160 Prouzel, France)
- Age at study initiation: no data
- Weight at study initiation: 2.7 +/- 0.01 kg
- Housing: individually in polystyrene cages
- Diet (e.g. ad libitum): Rabbits sustenance 112 C (UAR, 91360 Villemoisson/Orge, France)
- Water (e.g. ad libitum): filtered tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 50 +/- 20
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

not rinsed

Observation period (in vivo):

up to 72 h

Number of animals or in vitro replicates:

3

Details on study design:

Comment: not rinsed
A single dose of 0.1 ml of the test substance in its original form was instilled into the conjunctival sac of the left eye of 3 male New Zealand White rabbits. The eyes were not rinsed after instillation of the test substance. The ocular reactions were observed one hour, 24, 48 and 72 hours after the instillation. The mean score of the scores recorded after 24, 48 and 72 hours for each animal was calculated.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal: #1 and #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Basis:

animal: #1 and #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2 and#3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal: #1; #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Some slight conjunctival reactions (chemosis and redness: mean score <= 0.3 in all 3 animals) were observed after instillation of the test substance. No iridic irritation or corneal opacity was noted. No ocular reactions were observed after 48 and 72 hours.

Interpretation of results:

GHS criteria not met

Conclusions:

TPS 44 was considered as very slightly irritant when administered by ocular route in rabbits.

Executive summary:

The ocular irritation that could be induced by di-tert-butyl polisulfides (TPS 44) was evaluated in rabbits according to the recommendations of the OECD Guideline No. 405 and GLP. Having confirmed that TPS 44 did not have a very irritant or corrosive effect when administered by cutaneous route, a single dose of 0.1 ml of TPS 44 in its original form was instilled into the conjunctival sac of the left eye of 3 male New Zealand White rabbits. The eyes were not rinsed after instillation of the test substance. The ocular reactions were observed one hour, 24, 48 and 72 hours after the instillation. The mean score of the scores recorded after 24, 48 and 72 hours for each animal was calculated. Some slight conjunctival reactions (mean chemosis score of 0.3 in 2 animals and mean redness score of 0.3 in 2 animals) were observed after instillation of the test substance. No iridic irritation or corneal opacity was noted. No ocular reactions were observed after 48 and 72 hours. TPS 44 was considered as very slightly irritant when administered by ocular route in rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

In a key study, the potential of the registered substance to induce skin irritation was evaluated in rabbits according to OECD No. 404 and EC (2004/73/EC, B.4) guidelines and in compliance with the principles of GLP (Ollivier, 2006). The registered substance was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit. Since the test item was not severely irritant on this first animal, it was then applied for 4 hours to two other animals. A single dose of 0.5 mL of undiluted substance was applied to the closely-clipped skin of one flank. The registered substance was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the end of the observation period. The mean values of the scores for erythema and edema were calculated for each animal. After a 3-minute exposure (one animal), a very slight or well-defined erythema was noted from day 1 up to the end of the observation period (day 15). A very slight or slight edema was observed from day 3 up to day 9 and dryness of the skin was also recorded from day 5 up to the end of the observation period (day 15). After a 1-hour exposure (one animal), a very slight or well-defined erythema was noted from day 1 up to the end of the observation period (day 15). A very slight or slight edema was observed from day 4 up to day 9 and dryness of the skin was also recorded from day 6 up to the end of the observation period (day 15). After a 4-hour exposure (three animals), a very slight or well-defined erythema was noted in all animals from day 1 up to day 12, 13 or 15 (end of the observation period). A very slight or slight edema was observed in 2/3 animals from day 1 up to day 3 or 7 and dryness of the skin was also recorded in all animals between days 5 and 15 (end of the observation period). Mean scores over 24, 48 and 72 hours for each animal were 2.0, 2.0 and 2.0 for erythema and 0.0, 1.0 and 2.0 for edema.

In a supporting study, the cutaneous irritation that could be induced by the registered substance was evaluated in rabbits according to the OECD Guideline No. 404 and compliance to GLP (Clouzeau, 1992b). One male New Zealand White rabbit received on a 6 cm2 clipped area, a single dose of 0.5 ml of substance in its original form and prepared on a dry compress. The test substance was held in contact with the skin for 4 hours by means of a semi-occlusive dressing. The cutaneous reactions were observed one hour, 24, 48 and 72 hours after removal of the dressing and then daily in order to observe their reversibility or irreversibility. No residual test substance was observed after removal of the dressing. As the test substance was considered not very irritating, the first results were confirmed on 2 other animals. The mean value of the scores recorded after 24, 48 and 72 hours for each animal was calculated. Very slight and well-defined erythema were observed after application of the substance and reversed on day 7. No oedema was noted. The mean score for erythema was 0.3 in one animal and 2.0 in 2 animals. A dryness of the skin was observed between days 4 and 15.

Eye irritation

In a key study, the ocular irritation that could be induced by the registered substance was evaluated in rabbits according to the recommendations of the OECD Guideline No. 405 and compliance to GLP (Clouzeau, 1992c). Having confirmed that the registered substance was not very irritating or corrosive when administered by the cutaneous route, a single dose of 0.1 ml of undiluted substance was instilled into the conjunctival sac of the left eye of 3 male New Zealand White rabbits. The eyes were not rinsed after instillation of the test substance. The ocular reactions were observed one hour, 24, 48 and 72 hours after the instillation. The mean value of the scores recorded after 24, 48 and 72 hours for each animal was calculated. Some slight conjunctival reactions (mean chemosis score of 0.3 in 2 animals and mean redness score of 0.3 in 2 animals) were observed after instillation of the test substance. No iridic irritation or corneal opacity was noted. No ocular reactions were observed after 48 and 72 hours. The registered substance was considered as very slightly irritant when administered by ocular route in rabbits.

 

Respiratory irritation

 

No data are available to evaluate the respiratory irritation potential of  the registered substance. However, considering the low vapor pressure and the absence of significant irritation on the skin and eyes,  the registered substance is not considered as a potential irritant for the respiratory tract.

 

.

Justification for classification or non-classification

According to the criteria of EC regulation No 1272/2008, no classification is warranted for skin and eye irritation.