Pentapotassium bis(peroxymonosulphate) bis(sulphate)

Endpoint summary

Description of key information

KMPS triple salt was tested for skin sensitisation in a Local Lymph Node Assay (BrdU ELISA method) according to OECD guideline 442B. Topical application of test article (containing 2.86% of the impurity dipotassium peroxodisulphate, CAS 7727-21-1) at 0.1%, 0.25%, and 0.5% (w/v) in DMSO resulted in SI values less than 1.6 (SI < 1.6), and thus, the test article is not considered to be a dermal sensitiser.

 

Furthermore, KMPS triple salt (containing 2% of the impurity dipotassium peroxodisulphate, CAS 7727-21-1) was tested for skin sensitisation in a Guinea Pig Maximisation Test according to OECD guideline 406 and GLP. No skin sensitisation was observed after intradermal injection with 0.25%, topical induction with 20% and topical challenge with 5 and 2.5% of the test substance. Thus, the test substance was considered to be not skin sensitising.

 

Four other studies (two guinea pig maximisation tests and two Buehler tests similar to OECD guideline 406) containing unknown amounts of the impurity dipotassium peroxodisulphate (CAS 7727-21-1) were conducted. No sensitising activity was detected in any of these tests.

In addition epidemiological data from exposed workers showed no skin or respiratory sensitisation.

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:

weight of evidence

Study period:

1985-02-13 - 1985-05-29

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. During the rechallenge a high incidence of skin reactions was observed in both control and test group animals after application of distilled water only.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

no

Remarks:

, several quality assurance inspections were carried out.

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study as conducted before the LLNA became the standard method for testing skin sensitisation.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

not indicated

Route:

intradermal and epicutaneous

Vehicle:

other: distilled water

Concentration / amount:

Primary induction (intradermal): 1% w/v in distilled water and in Freunds Complete adjuvant, respectively
Secondary Induction (epicoutaneous): 0.6 ml 15% w/v KMPS triple salt in distilled water
First Challenge: 0.2 ml 6% w/v KMPS triple salt in distilled water
Second Challenge: 0.2 ml 3% w/v KMPS triple salt in distilled water

Route:

epicutaneous, occlusive

Vehicle:

other: distilled water

Concentration / amount:

Primary induction (intradermal): 1% w/v in distilled water and in Freunds Complete adjuvant, respectively
Secondary Induction (epicoutaneous): 0.6 ml 15% w/v KMPS triple salt in distilled water
First Challenge: 0.2 ml 6% w/v KMPS triple salt in distilled water
Second Challenge: 0.2 ml 3% w/v KMPS triple salt in distilled water

No. of animals per dose:

ten males, ten females,
controls: ten males, ten females

Details on study design:

not indicated

Challenge controls:

not indicated

Positive control substance(s):

not specified

Positive control results:

not indicated

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

other: first challenge, 1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

6% w/v KMPS triple salt

No. with + reactions:

9

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: first challenge, 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 6% w/v KMPS triple salt. No with. + reactions: 9.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: first challenge, 2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

6% w/v KMPS triple salt

No. with + reactions:

1

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: first challenge, 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 6% w/v KMPS triple salt. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: first challenge, 1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

6% w/v KMPS triple salt

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: first challenge, 1st reading. . Hours after challenge: 24.0. Group: other: concurrent control. Dose level: 6% w/v KMPS triple salt. No with. + reactions: 5.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: first challenge, 2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

6% w/v KMPS triple salt

No. with + reactions:

1

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: first challenge, 2nd reading. . Hours after challenge: 48.0. Group: other: concurrent control. Dose level: 6% w/v KMPS triple salt. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: first challenge, 1st and 2nd reading

Group:

test chemical

Dose level:

distilled water

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: other: first challenge, 1st and 2nd reading. Group: other: test group. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Key result

Reading:

other: first challenge, 1st and 2nd reading

Group:

negative control

Dose level:

distilled water

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: other: first challenge, 1st and 2nd reading. Group: other: concurrent control group. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Key result

Reading:

other: rechallenge, 1st and 2nd reading

Group:

test chemical

Dose level:

3% w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: other: rechallenge, 1st and 2nd reading. Group: test group. Dose level: 3% w/v KMPS triple salt. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Key result

Reading:

other: rechallenge, 1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

3% w/v KMPS triple salt

No. with + reactions:

1

Total no. in group:

20

Clinical observations:

only slight, patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: rechallenge, 1st reading. . Hours after challenge: 24.0. Group: other: concurrent control. Dose level: 3% w/v KMPS triple salt. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: only slight, patchy erythema.

Key result

Reading:

other: rechallenge, 2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

3% w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: other: rechallenge, 2nd reading. . Hours after challenge: 48.0. Group: other: concurrent control. Dose level: 3% w/v KMPS triple salt. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Key result

Reading:

other: rechallenge: 1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

distilled water

No. with + reactions:

9

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: Reading: other: rechallenge: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: distilled water. No with. + reactions: 9.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: rechallenge: 2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

distilled water

No. with + reactions:

1

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: Reading: other: rechallenge: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: distilled water. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: rechallenge, 1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

distilled water

No. with + reactions:

3

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: rechallenge, 1st reading. . Hours after challenge: 24.0. Group: other: concurrent control. Dose level: distilled water. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Key result

Reading:

other: rechallenge, 2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

distilled water

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

only slight or moderate patchy erythema

Remarks on result:

other: see Remark

Remarks:

Reading: other: rechallenge, 2nd reading. . Hours after challenge: 48.0. Group: other: concurrent control. Dose level: distilled water. No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: only slight or moderate patchy erythema.

Primary irritation screen

Intradermal injection

Administration of 100% w/v KMPS triple salt in Freunds Complete Adjuvant proved to be impossible by injection into the dermis. Sites of intradermal injection of formulations incorporating 3% w/V KMPS triple salt in distilled water or Freunds Complete Adjuvant showed dermal changes including loss of elasticity or eschar formation which were considered to reflect substantial damage of the treated tissue. Reactions to intradermal injection of 1% w/v KMPS triple salt in either vehicle were blanching, slight or moderate erythema and or discolouration.

Occluded topical application (first phase)

Occluded topical application of 20%, 30% and 40% w/v KMPS triple salt in distilled water caused discolouration and reduced dermal pliability or eschar formation and yet treatment with 10% w/v KMPS triple salt in distilled water failed to elicit any dermal change other than pitting of the surface on one of the test sites.

Occluded topical application (second phase)

There were no irritation responses or dermal changes at the sites of occluded topical application of 3% or 6% w/v KMPS triple salt in distilled water. Formulations incorporating 12% or 20% w/v KMPS triple salt in distilled water elicit slight or moderate erythema and, in some cases, discolouration of the treated skin.

Induction

Intradermal injection

Administration of 1% w/v KMPS triple salt in distilled water or the adjuvant or of the vehicles alone or in combination, caused dermal changes such as discolouration and irritation reactions not exceeding slight erythema.

Occluded topical application

Three control males showed slight, confluent or moderate, patchy erythema 4 hours after removal of occlusive dressings which had applied distilled water to their dorsum. Contemporaenous treatment of test group animals with 15% w/v KMPS triple salt in distiled water elicited similar erythematous responses from eight guinea-pigs, but coagulated blood from injection sites disturbed during the occlusion procedure had partially obscured the dermal test sites of more than one-half of he test group animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The incidence of significant erythematous reations to challenge with 3% w/v KMPS triple salt in distilled water was not greater among animals previously subjected to induction procedures involving repeated administration of the test material than among animals of a comtemporaneous control group.
Under the conditions of this study, it was concluded that repeated administration of KMPS triple salt has failed to cause delayed contact hypersensitivity in guinea-pigs.

Executive summary:

Materials and methods

The potential of KMPS triple salt to cause delayed contact hypersinsitivity in guinea-pigs was assessed by the Magnusson-Kligman Maximisation Test.

 

The shaven dorsum of ten male and ten female Dunkin-Hartley guinea-pigs were subjected to intradermal injections of Freunds Complete Adjuvant, 1% w/v KMPS triple salt in distilled water and 1% w/v KMPS triple salt in the adjuvant on Day 1. Seven days later the same area of skin was treated by topical application of 15% w/v KMPS triple salt in distilled water and the test site was covered by an occlusive dressing for48 hours. the same induction procedures were carried out on a contemporaneous control group, except that the test material was replaced by vehicle in all doses.

On day 22 all animals were challenged by occluded application of distilled water to the left flank and 6% w/v KMPS triple salt in distilled water to the right flank. The occlusive dressings were removed on the following day and the condition of the test sites was assessed approximately 24 and 48 hours later.

The result obtained at first challenge was tested by repeating the procedure on Day 29. On this occasion the vehicle was applied to a naive site on the right flank and 3% w/v KMPS triple salt in distilled water was applied to the left flank.

 

Results and discussion

First challenge application of distilled water failed to elicit any erythematous responses.

Contemporaneous treatment of the opposite flank with 6% w/v KMPS triple salt in distilled water caused slight or moderate patchy erythema in five controls and nine test group animals examined 24hours after challenge and in single test and control group animals examined 48 hours after challenge.

The test sites subjected to the second challenge with the vehicle showed slight or moderate patchy erythema in three control and nine test group animals at the first examination and in two control and one test animal at the second examination.

Slight, patchy erythema affected a single control male 24 hours after second challenge application of 3% w/v KMPS triple salt in distilled water in the first reading and disappeared in the 2nd reading.

Reference 2

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1985-02-13 - 1985-04-28

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. The modified Buehler test is not considered to be sensitive enough when compared to the GPMT.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. The modified Buehler test is not considered to be sensitive enough when compared to the GPMT.

Principles of method if other than guideline:

The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. The modified Buehler test is not considered to be sensitive enough when compared to the GPMT.

GLP compliance:

no

Remarks:

several quality assurance inspections were carried out.

Type of study:

Buehler test

Justification for non-LLNA method:

The study as conducted before the LLNA became the standard method for testing skin sensitisation.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

not indicated

Route:

epicutaneous, occlusive

Vehicle:

other: distilled water

Concentration / amount:

Primary irritation screen: 25%, 50%, 100%, 200% w/v
Induction: 25% w/v
Challenge: 6%

Route:

epicutaneous, occlusive

Vehicle:

other: distilled water

Concentration / amount:

Primary irritation screen: 25%, 50%, 100%, 200% w/v
Induction: 25% w/v
Challenge: 6%

No. of animals per dose:

ten males, ten females,
controls: five males, five females

Details on study design:

not indicated

Challenge controls:

not indicated

Positive control substance(s):

not specified

Positive control results:

not indicated

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.4 ml 6% w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

2 animals with two white raised marks across site

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.4 ml 6% w/v KMPS triple salt. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: 2 animals with two white raised marks across site.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.4 ml 6% w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

one animal with very faint erythema, ususally non-confluent

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.4 ml 6% w/v KMPS triple salt. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: one animal with very faint erythema, ususally non-confluent.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

one animal scratched

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: concurrent control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: one animal scratched.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

one animal scratched

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: concurrent control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: one animal scratched.

Primary irritation screen:

Single occluded application of 50%, 100% or 200% w/v KMPS triple salt in distilled water caused loss of elasiticity and darkening of all dermal test sites treated with these formulations. This was considered to indicate in-depth damage of the treated skin. Areas of the dorsum subject to treatment with 25% w/v KMPS triple salt in distilled water showed slight discolouration or faint erythema and discolouration or, in one case, less of an area of epidermis leading to eschar formation.

Bodyweight and general condition:

All animals achieved anticipated bodyweight gains and remained in overt good health throughout the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The incidence of significant erythematous responses to challenge with 6% w/v KMPS triple salt in distilled water was not higher among animals previously subjected to repeated occluded topical applications of 25% w/v KMPS triple salt in distilled water than among animals of the contemporaneous control group. Accordingly, it was concluded that, under the conditions of this study, repeated occluded dermal applications of 25% w/v KMPS triple salt in distilled water failed to cause delayed contact hypersensitivity in guinea-pigs.

Executive summary:

Materials and methods:

The potential of KMPS triple salt to cause delayed contact hypersinsitivity in guinea-pigs was assessed by a modified version of the method of Buehler (1965).

The shaven dorsum of ten male and ten female albino guinea-pigs was subject to a three phase induction procedure by occluded application of 25% w/v KMPS triple salt in distilled water for periods of appproximately six hours on Days 1, 8 and 15. A control group of five male and five female naive guinea-pigs was selected from the same stock used to supply the test group but remained untreated during the induction phase of the study.

On Day 29 all test and control animals were challenged by six hours occluded application of 6% w/v KMPS triple salt in distilled water to an area of the dorsum not treated during the induction phase of the study. Dermal responses to the challene procedure were assessed 24 and 48 hours after application of the occlusive dressings.

 

Results and discussion:

Induction:

First induction elicited faint erythema from six guinea-pigs and moderate erythema from a seventh animal. All dermal test sites showed slight discolouration on the day after treatment.

Approximately three-quarters of the test group had developed moderate erythema and very minor areas of superficial eschar on the day after the second induction procedure. Other animals generally showed a lesser erythematous reaction associated with minor superficial eschar formation. In addition, the treatment had a depilatory effect on all dermal test sites and caused discolouration or the formation of larger areas of eschar (appproximately 5 x 5 mm) in several animals.

The effects of third induction were similar to those following second induction except for reduced incidence of minor superficial eschar formation and increased incidence of the formation of larger areas of eschar.

 

Challenge:

Challenge applications of 6% w/v KMPS triple salt in distilled water to naive dermal test sites elicited no erythematous responses from test or control group guinea-pigs excepting very faint erythema apparent on one test male 48 hours after challenge.

Reference 3

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1985-02-13 - 1985-04-28

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. The modified Buehler test is not considered to be sensitive enough when compared to the GPMT.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. The modified Buehler test is not considered to be sensitive enough when compared to the GPMT.

Principles of method if other than guideline:

The study was not performed according to GLP. The identity and purity of the test material was not specified in the study report. The modified Buehler test is not considered to be sensitive enough when compared to the GPMT.

GLP compliance:

no

Remarks:

Not required in the period, when the study was performed

Type of study:

Buehler test

Justification for non-LLNA method:

The study as conducted before the LLNA became the standard method for testing skin sensitisation.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

not indicated

Route:

epicutaneous, occlusive

Vehicle:

other: distilled water

Concentration / amount:

Primary irritation screen: 25%, 50%, 100%, 200% w/v
Induction: 25% w/v
Challenge: 6% w/v

Route:

epicutaneous, occlusive

Vehicle:

other: distilled water

Concentration / amount:

Primary irritation screen: 25%, 50%, 100%, 200% w/v
Induction: 25% w/v
Challenge: 6% w/v

No. of animals per dose:

ten males, ten females,
controls: five males, five females

Details on study design:

not indicated

Challenge controls:

not indicated

Positive control substance(s):

not specified

Positive control results:

not indicated

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.4 ml of 6% w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

1 male scratched

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.4 ml of 6% w/v KMPS triple salt . No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: 1 male scratched.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.4 ml of 6% w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

1 male scratched, 2 females very faint erythema, usually non-confluent

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.4 ml of 6% w/v KMPS triple salt . No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: 1 male scratched, 2 females very faint erythema, usually non-confluent.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0.4 ml of 6 % w/v KMPS triple salt

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: concurrent control. Dose level: 0.4 ml of 6 % w/v KMPS triple salt. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0.4 ml of 6 % KMPS triple salt

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: concurrent control. Dose level: 0.4 ml of 6 % KMPS triple salt. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Primary irritation screen:

Single occluded application of 50%, 100% or 200% w/v KMPS triple salt in distilled water caused eschar formation or loss of elasiticity and darkening of all dermal test sites treated with these formulations. This was considered to reflect in-depth injury of the treated skin.

Areas of the dorsum subject to treatment with 25% w/v KMPS triple salt in distilled water showed slight discolouration or faint erythema and discolouration.

 

Body weight and general condition:

All animals achieved anticipated bodyweight gains and remained in overt good health throughout the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The incidence of significant erythemaous responses to challenge with 6% w/v KMPS triple salt in distilled water was not higher among animals previously subjected to repeated occluded topical applications of 25% w/v KMPS triple salt in distilled water than among animals of the contemporaneous control group. Accordingly, it was concluded that, under the conditions of this study, repeated occluded dermal applications of 25% w/v KMPS triple salt in distilled water failed to cause delayed contact hypersensitivity in guinea-pigs.

Executive summary:

Materials and methods

The potential of KMPS triple salt to cause delayed contact hypersenitivity in guinea-pigs was assessed by a modified version of the method of Buehler (1965).

The shaven dorsum of ten male and ten female albino guinea-pigs was subject to a three phase induction procedure by occluded application of 25% w/v KMPS triple salt in distilled water for periods of approximately six hours on Days 1, 8 and 15. A control group fo five male and five female naive guinea-pigs was selected from the same stock used to supply the test group but remained untreated during the induction phase of the study.

On day 29 all test control animals were challenged by six hours occluded application of 6% w/v KMPS triple salt in distilled water to an area of the dorsum not treated during the induction phase of the study. Dermal responses to the challenge procedure were asessed 24 and 48 hours after application of the occlusive dressing.

Results and discussion

Induction:

The majority of dermal test sites examined on the day after first induction were slightly discoloured. One male and two female guinea-pigs showed faint erythema as well as, or in place of discolouration.

Second induction caused slight, moderate or, in one case, severe erythema of all treated areas of dorsum. Treatment usually had a depilatory effect upon the treated skin and the majority of test sites developed areas of discolouration or large areas (approximately 5 x 5 mm) of eschar formation. Similar dermal changes followed third induction although eschar was normally apparent only as a superficial layer affecting very minor areas of the test sites.

Challenge:

Challenge application of 6% w/v KMPS triple salt in distilled water to naive dermal test sites elicited no erythematous responses from test and control group animals, excepting very faint erythema apparent on two test group guinea-pigs 48 hours after challenge.

Reference 4

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1991-04-03 - 1992-01-13

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The identity and purity of the test material was not specified in the study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study as conducted before the LLNA became the standard method for testing skin sensitisation.

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

not indicated

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Concentration / amount:

Induction
- Intradermal: 0.05%
- Epicutaneous: 10%

Challenge
- Epicutaneous: 3%

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

Induction
- Intradermal: 0.05%
- Epicutaneous: 10%

Challenge
- Epicutaneous: 3%

No. of animals per dose:

Control group 1: 6 animals (identical treatment of animals of the test substance group)
Control group 2: 6 animals (as the result of the first challenge was unequivocal, a second challenge was not performed. The animals of the control group 2 were left untreated)
Treatment group: 10 animals

Details on study design:

not indicated

Challenge controls:

not indicated

Positive control substance(s):

not specified

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

3%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

no systemic toxic effects

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 3%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no systemic toxic effects.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

3%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

no systemic toxic effects

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no systemic toxic effects.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

6

Clinical observations:

none observed

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: none observed.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

6

Clinical observations:

none oberved

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: none oberved.

Interpretation of results:

GHS criteria not met

Conclusions:

The study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability). KMPS triple salt has no sensitizing properties on the skin of the guinea pig. No classification as sensitiser is necessary according to Regulation 1272/2008/EC (CLP).

Executive summary:

Materials and methods

KMPS triple salt was applied to the skin of guinea pigs to determine its sensitizing properties by the maximization test method. The test substance was dissolved in physiological saline solution 0.9%. The concentrations were 0.05% (intradermal) or 10% (epidermal) during induction and 3 % (epidermal) during challenge.

 

Results and discussion

Following epidermal challenge, none of the animals of the test substance group showed any findings at the treated skin sites.

None of the animals of the control group showed changes at the exposed skin areas. Systemic toxic effects could not be detected. The general condition and body development of the test animals were not affected.

Reference 5

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2001-03-23 - 2001-10-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

July 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

September 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Version / remarks:

November 1982, revised August 1998

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study as conducted before the LLNA became the standard method for testing skin sensitisation.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire; UK
- Age at study initiation: 4 - 7 weeks
- Weight at study initiation: 534 – 625 g

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

a) intradermal induction:
FCA (Freund’s Complete Adjuvant) in water (1:1), test substance diluted to 0.25 % (w/v) with water, test substance diluted to 0.25 % (w/v) by a 1:1 mixture (v/v) of FCA and water
b) topical induction: application of 20 % (w/v) test material in water
c) challenge: application of 5 and 2.5 % (w/v) test material in water

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

a) intradermal induction:
FCA (Freund’s Complete Adjuvant) in water (1:1), test substance diluted to 0.25 % (w/v) with water, test substance diluted to 0.25 % (w/v) by a 1:1 mixture (v/v) of FCA and water
b) topical induction: application of 20 % (w/v) test material in water
c) challenge: application of 5 and 2.5 % (w/v) test material in water

No. of animals per dose:

Pretest: 10 animals (2 for intradermal and 8 for topical application)
Main test: 15 animals (5 control and 10 test group animals)

Details on study design:

ADMINISTRATION/EXPOSURE

Induction schedule:
day 1: intradermal induction of a 0.25 % test item dilution in water, 1:1 mixture of FCA/water and at 0.25 % test item in a 1:1 mixture of FCA/water
day 7: clipping and shaving
day 8: topical induction with 20 % Oxone Monopersulfate Compound in water
day 22: topical challenge using 5 and 2.5 % Oxone Monopersulfate Compound in water

Way of Induction:
Intradermal injection followed by topical application one week later

Concentrations used for induction:
a) intradermal:
0.1 mL of 0.25 % Oxone Monopersulfate Compound (in water), 0.1 mL 1:1 mixture of FCA/water and 0.1 mL 0.25 % Oxone Monopersulfate Compound in a 1:1 mixture of FCA/water
b) topical:
20 % (w/v) Oxone Monopersulfate Compound in water

Concentration Freunds Complete Adjuvant (FCA):
intradermal induction: 0.1 mL injected
injection site 1: FCA diluted 1:1 with water
injection site 2: 0.25 % (w/v) Oxone Monopersulfate Compound in water
injection site 3: 0.25 % (w/v) Oxone Monoperulfate Compound in a 1:1 mixture of FCA and water

Challenge schedule: day 22

Concentrations used for challenge: 5 and 2.5 % Oxone Monopersulfate Compound in water

Rechallenge: none

Scoring schedule:
Dermal reactions for test and control animals to the intradermal injections were recorded 24 hours following the injections to the induction. Topical application were recorded on removal of the bandages.
Challenge sites were evaluated app. 24 and 48 hours after removal of the patches.

Removal of the test substance:
The test substance was removed 48 hours after topical induction application and 24 hours after challenge application.


EXAMINATIONS

Pilot study:
The intradermal and topical irritancy of a range of dilutions of the test substance was investigated to identify where possible
(a) the minimum irritant test substance concentrations suitable for the induction phase of the main study and
(b) a maximum non-irritant concentration by the topical route and a dilution of this for the challenge phase.

The animals of the topical irritancy investigations were pre-treated with an intradermal injection of Freund’s Complete Adjuvant, 50:50 with sterile water, approximately one week prior to the start of the preliminary investigations.
Intradermal injections: Intradermal injections (0.1 mL/site) were made into the clipped and shaved flank of two guinea pigs, using a range of concentration (0.1 to 10 % w/v) of Oxone Monopersulfate® Compound in sterile water. The resulting dermal responses were assed approximately 24 and 72 h later.
Topical applications: Patches were satured (volume approximately 0.2 mL/patch) with a range of concentrations (0.01 to 100 % w/v) of Oxone Monopersulfate® Compound in sterile water and applied to eight guinea pigs. The dressings were removed after approximately 24 h and reaction sites were assessed for erythema and oedema. Further examination of the sites was carried out approximately 24 and 48 h after removal of the dressings.

Erythema and oedema were assessed by using the numerical grading system according to Draize.

Challenge controls:

Control were treated topically in the same manner than the test group animals by applying 2.5 and 5% (w/v) solution of the compound in water to the clipped flanks.

Positive control substance(s):

yes

Remarks:

The sensitivity of the guinea-pig strain used is checked periodically with hexyl cinnamic aldehyde (HCA). The results of the most recent test were used as positive control.

Positive control results:

not indicated

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

2.5 and 5 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none observed

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 2.5 and 5 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none observed.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

2.5 and 5 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none observed

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 2.5 and 5 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none observed.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

2.5 and 5 %

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none observed

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 2.5 and 5 %. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none observed.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

2.5 and 5 %

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none observed

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 2.5 and 5 %. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none observed.

Key result

Reading:

1st reading

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

2nd reading

Group:

positive control

Remarks on result:

not measured/tested

Results of pilot studies

Intradermal Induction:

The intradermal application of 1.0 to 10.0 % (w/v) of KMPS triple salt in water produced necrosis in both animals tested. In case of the induction by 0.5 % (w/v) solution necrosis was observed in one guinea pig only. The highest concentration applied which did not cause necrosis but well defined irritations was 0.25 % KMPS triple salt in water.

 

Topical Induction:

20 % (w/v) of the test substance in water were chosen for topical induction. This was the highest concentration that produced some irritation but did not adversely affect the animals. 5 and 2.5 % (w/v) of KMPS triple salt in water were selected for the topical challenge. This was the highest concentration which gave no rise to irritating effects.

 

 

Results of main test

24 and 48h after first challenge

Intradermal injections: Necrosis was recorded at sites receiving FCA in test and control animals. No irritation was seen in the test animals at sites receiving KMPS triple salt, 0.25 % (w/v) in water for irrigation and no irritation was observed in control animals receiving water for irrigation (please refer to table A6.1.5/01-1).

Topical application: Well defined to moderate erythema, accompanied by blanching of the dose site, was observed in all test animals following topical application with KMPS triple salt, 20 % (w/v) in water for irrigation. No erythema was seen in the

control group (please refer to table 1).

Challenge: There were no dermal reactions seen in any of the test or control animals. Consequently all ten test animals gave negative responses (please refer to table 2).

 

Other findings

None of the animals of the control or test group showed clinical signs of toxicity and no mortalities were observed. There were no effects on body weight of animals of the main study.

 

Overall result

None of the animals demonstrated any evidence for an allergic reaction. KMPS triple salt is therefore not considered to be a potential skin sensitiser.

 

Table 1: Dermal reactions observed after each induction

Group	Animal number	Intradermal injections			Topical application
		Site number
		1	2	3
Control	365	N	0	N	0
"	366	N	0	N	0
"	367	N	0	N	0
"	368	N	0	N	0
"	369	N	0	N	0
-	-	-	-	-	-
Test	370	N	0	N	2#
"	371	N	0	N	2#
"	372	N	0	N	2#
"	373	N	0	N	2#
"	374	N	0	N	2#
"	375	N	0	N	2#
"	376	N	0	N	2#
"	377	N	0	N	2#
"	378	N	0	N	2#
"	379	N	0	N	2#

 

 

Intrademal injections:

 

Site No. 1: 0.1 mL FCA/water 1:1

Site No. 2: 0.1 mL water (control)

0.1 mL KMPS triple salt (0.25 % w/v in water) (test)

Site No. 3: 0.1 mL FCA/water 1:1 (control)

0.1 mL KMPS triple salt (0.25 % w/v in FCA/water 1:1) (test)

 

N Necrosis

0 No irritation

1 Slight irritation

2 Well-defined irritation

3 Moderate irritation

4 Severe irritation

 

 

Topical application:

 

Control: water for irrigation

Test: KMPS triple salt, 20 % w/v in water for irrigation

 

0 No erythema

1 Slight erythema

2 Well defined erythema

3 Moderate erythema

4 Severe erythema

# Blanching of the dose site

 

Table 2a: Dermal reactions observed after the challenge with KMPS triple salt
Freund’s treated controls

Guinea pig Number	E = Erythema O = Oedema	Score
		24 hours		48 hours
-	-	Aa	Pb	Aa	Pb
365	E O	0 0	0 0	0 0	0 0
366	E O	0 0	0 0	0 0	0 0
367	E O	0 0	0 0	0 0	0 0
368	E O	0 0	0 0	0 0	0 0
369	E O	0 0	0 0	0 0	0 0

a    A: anterior site, exposed to KMPS triple salt, 5 % w/v in sterile water

b    P: posterior site, exposed to KMPS triple salt, 2.5 % w/v in sterile water

 

 

Table 2b: Dermal reactions observed after the challenge with KMPS triple salt

Guinea pig Number	E = Erythema O = Oedema	Score				Resultsa
		24 hours		48 hours
-	-	Ab	Pc	Ab	Pc	-
370	E O	0 0	0 0	0 0	0 0	-
371	E O	0 0	0 0	0 0	0 0	-
372	E O	0 0	0 0	0 0	0 0	-
373	E O	0 0	0 0	0 0	0 0	-
374	E O	0 0	0 0	0 0	0 0	-
375	E O	0 0	0 0	0 0	0 0	-
376	E O	0 0	0 0	0 0	0 0	-
377	E O	0 0	0 0	0 0	0 0	-
378	E O	0 0	0 0	0 0	0 0	-
379	E O	0 0	0 0	0 0	0 0	-

a      (+): positive; (-): negative; (±): inconclusive

b    A: anterior site, exposed to KMPS triple salt, 5 % w/v in sterile water

c    P: posterior site, exposed to KMPS triple salt, 2.5 % w/v in sterile water

Interpretation of results:

GHS criteria not met

Conclusions:

The study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability). KMPS triple salt (containing 2% of the impurity dipotassium peroxodisulphate, CAS 7727-21-1) was tested for skin sensitisation in the guinea pig maximisation test. No sensitisation was detected in this test. KMPS triple salt has not to be classified as a skin sensitiser according Regulation 1272/2008/EC (CLP).

Executive summary:

Materials and methods

The skin sensitisation potential of KMPS triple salt (containing 2% of the impurity dipotassium peroxodisulphate, CAS 7727-21-1) was investigated in the guinea pig maximisation test according to EU Guideline B.6 (Skin Sensitisation). A control and test group consisting of 5 and 10 male albino guinea pigs each were used for the test. Suitable concentrations for intradermal injections as well as topical induction and challenge application were selected in a pretest. Animals were intradermally induced on day 1 and topical induced on day 8. Animals were challenged on day 22. All animals were observed for clinical signs and mortality. No necropsies on control or test group animals were performed on termination of the study.

 

Results and discussion

Following intradermal injection necrosis was recorded at sites receiving FCA in test and control animals. No irritation was seen in the test animals at sites receiving KMPS triple salt, 0.25 % (w/v) in water for irrigation and no irritation was observed in control animals receiving water for irrigation.

Following topical application with KMPS triple salt, 20 % (w/v) in water for irrigation, well defined to moderate erythema, accompanied by blanching of the dose site, was observed in all test animals. No erythema was seen in the control group.

Following challenge application no dermal reactions were seen in any of the test or control animals. Consequently all ten test animals gave negative responses.

Reference 6

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2020-04-21 to 2020-08-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 442B (Skin sensitisation: Local Lymph Node Assay: BrdU-ELISA or –FCM)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA): BrdU-ELISA

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Jackson Laboratories (Bar Harbor, ME)
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: Approx. 8 weeks
- Weight at study initiation: 21.4-22.7 (preliminary dermal irritation screen), 19.6-23.0 (Quantitative Irritation Test) , 22.1-27.1 (main study)
- Housing: One per cage in suspended wire-bottom cages
- Diet: Ad libitum (PMI Rodent Chow)
- Water: Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not specified, but room was temperature-controlled using automatic recording devices.
- Humidity (%): Not specified, but room was humidity-controlled using automatic recording devices.
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): 12/12

- IN-LIFE DATES: From: Day 1 to day 6

Vehicle:

dimethyl sulphoxide

Concentration:

Range finding test: 5%, 10%, 25% (w/v) in DMSO
Quantitative Irritation Test: 0.05%, 0.1%, 0.25%, 0.5%, 1.0%, and 2.5% (w/v) in DMSO
Main test: 0.1%, 0.25% and 0.5% (w/v) in DMSO

The concentrations of the main tests were chosen based on the preliminary tests, such that the
maximum concentration tested avoided both overt systemic toxicity and more-than-moderate local dermal irritation.

No. of animals per dose:

Range finding test: Two female animals per group
Quantitative Irritation Test: Two female animals per group
Main test: Five female animals per group

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility: The test article was tested for solubility at 50% (w/v) in acetone:olive oil (4:1, AOO), dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), 2-butanone, and propylene glycol (PG). The test substance was only soluble in DMSO (opaque, off-white liquid)
- Irritation: Flaking skin was observed in all animals in the range finding test (5-25% (w/v) KMPS) and in animals treated with 0.25-2.5% (w/v) KMPS in the Quantitative Irritation Test.
- Systemic toxicity: No clinical signs were observed.
- Ear thickness measurements: Ear thickness was measured on day 1, 3 and 6. The ear thickness was >25% at all three doses in the range finding test. The ear thickness was >25% at 1.0% and 2.5% (w/v) Quantitative Irritation Test.
- Erythema scores: Mean erythema scores were 0.3-0.8 in the range finding test (5-25% w/v KMPS) and 0.3 (0.5% w/v KMPS), 0.5 (1.0% w/v KMPS) and 0.3 (2.5% w/v KMPS) in the Quantitative Irritation Test.

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: A test article is considered to have a positive response if treatment results in a 1.6-fold or greater increase in the mean LNC proliferation (BrdU ELISA OD values) relative to that obtained for the vehicle control. Therefore, a test article that yields an SI value of 1.6 or more is characterized as a sensitising material.

TREATMENT PREPARATION AND ADMINISTRATION: 0.015 g of the test article was brought to a total volume of 3000 μL with DMSO and mixed to yield a 0.5% (w/v) formulation. Additional
formulations to 0.25% and 0.1% (w/v) were made in DMSO. Five groups of CBA/J mice (five animals per group) were treated by topical application to the dorsum of each ear once daily for three consecutive days. The test article was spread over the entire dorsal surface of the ear using a positive displacement pipette to deliver 25 μL/ear.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Statistical evaluation of the calculated SI values was made by ANOVA followed by the Student-Newman- Keuls test, using InStat™ (version 3.06, 32 bit for Windows; GraphPad Software, San Diego, CA). However, determination of test article-induced sensitisation was based on the criterion that a test article that yields an SI value of 1.6 or more is characterized as a sensitiser.

Positive control results:

No clinical signs were observed after application of the positive control (25% HCA). Erythema scores were 0.9 (day 2), 1.3 (day 3), 1.6 (day 4), 1.4 (day 5) and 0.8 (day 6). An increase in ear thickness of >25% was observed on day 6. Furthermore, enlargement of the lymph nodes was observed at necropsy. Based on experience at the testing facility, this occurs in the majority of studies when using 25% HCA in the vehicle DMSO.

Key result

Parameter:

SI

Value:

1

Variability:

SD: 0.4

Test group / Remarks:

0.5% (w/v) KMPS

Key result

Parameter:

SI

Value:

1.3

Variability:

SD: 0.2

Test group / Remarks:

0.25% (w/v) KMPS

Key result

Parameter:

SI

Value:

1

Variability:

SD: 0.6

Test group / Remarks:

0.1% (w/v) KMPS

Key result

Parameter:

SI

Value:

3.1

Variability:

SD: 0.4

Test group / Remarks:

Positive control group (25% HCA)

Key result

Parameter:

SI

Value:

1

Variability:

SD: 0.1

Test group / Remarks:

Vehicle control group (DMSO)

Cellular proliferation data / Observations:

CELLULAR PROLIFERATION DATA
Visually larger lymph nodes compared to the vehicle control were observed in the positive control group. Appearance of the lymph nodes was normal in the negative (vehicle) control group and in all test item treated groups. Significant lymphoproliferative response (SI ≥ 1.6) was only observed for the positive control (SI = 3.1).

DETAILS ON STIMULATION INDEX CALCULATION: See "Any other information on materials and
methods incl. tables"

EC1.6 CALCULATION: The EC1.6 could not be calculated in this study because no test article treatment produced a mean SI value greater than 1.6.

CLINICAL OBSERVATIONS: No clinical signs were observed during treatment. Partially chewed food was observed in 1/5 animals in the 0.5% (w/v) KMPS group on day 5 and 6.

BODY WEIGHTS: Body weight changes were normal.

SIGNS OF TOXICITY: No to very slight erythema in the 0.25% and 0.5% (w/v) test article-treated groups were observed (score range: 0.1 - 0.6). None of the test article-treated groups had ear swelling that resulted in more-than-moderate dermal irritation (range: 5.0-10.0% increase)

Interpretation of results:

GHS criteria not met

Conclusions:

In a Local Lymph Node Assay (BrdU ELISA method) according to OECD guideline 442B, topical application of test article (containing 2.86% of the impurity dipotassium peroxodisulphate, CAS 7727-21-1) at 0.1%, 0.25%, and 0.5% (w/v) in DMSO resulted in SI values less than
1.6 (SI < 1.6), and thus, the test article is not considered to be a dermal sensitiser.

Executive summary:

In a Local Lymph Node Assay according to OECD guideline 442B and GLP, the sensitising potential of the topically applied test item (containing 2.86% of the impurity dipotassium peroxodisulphate, CAS 7727-21-1) was determined utilizing the BrdU ELISA method.

 

The test article was tested for solubility at 50% (w/v) in acetone:olive oil (4:1, AOO), dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), 2-butanone, and propylene glycol (PG). Test article solutions could not be made in AOO, DMF, 2-butanone, or PG. A soluble opaque off-white liquid was achieved with DMSO. Based on the solubility results, DMSO was chosen as the vehicle for the study. A preliminary dermal irritation screen was conducted with three groups (two animals per group) of healthy female CBA/J mice to determine the concentrations of the test article to be used in the main Local Lymph Node study. Three concentrations of the test article were tested: 5%, 10%, and 25% (w/v) in DMSO. The results of this preliminary screen indicated that all test article treatments resulted in dermal irritation and increased ear thickness (over 25%). Therefore, a quantitative irritation test (QIT) was performed, with six dose groups of two animals each, at test article concentrations of 0.05%, 0.1%, 0.25%, 0.5%, 1.0%, and 2.5% (w/v). The 1.0% and 2.5% (w/v) test article treatments resulted in dermal irritation and an increase in ear thickness. Test article residue at the dose site was observed in the 10% and 25% (w/v) test article-treated groups; however, this did not interfere with ear measurements. Based on these results, test article concentrations of 0.1%, 0.25%, and 0.5% (w/v) in DMSO were chosen for the main Local Lymph Node assay.

 

The main test was conducted with five separate groups of healthy female CBA/J mice (five animals per group). Three groups were treated with increasing concentrations of the test article. A vehicle control group was treated with DMSO and another group was treated with the positive control, alphahexylcinnamaldehyde in DMSO (25% HCA), in the exact same manner. The test article, vehicle control, and positive control solutions were administered by topical application to the dorsum of each ear, once daily for three consecutive days. All animals in the study were observed once daily throughout the study for clinical signs, either of local irritation at the application site or systemic toxicity, and for mortality. Body weights were measured on Day 1 prior to first dose and on Day 6 prior to euthanasia. The mice in the main test were given an intraperitoneal injection of the thymidine analog 5-bromo-2’-deoxy-uridine (BrdU) approximately 24 hours prior to euthanasia. After euthanasia, the auricular lymph nodes were isolated; single-cell suspensions of lymph node cells (LNC) were generated, then aliquots were permeabilized and denatured in triplicate in a 96-well microplate for analysis by ELISA for BrdU incorporation. The Stimulation Index (SI) was calculated by dividing the mean optical density of the LNC suspension for each animal by the mean optical density of the vehicle control group LNC suspension. The mean SI and standard deviation were calculated for each group from the individual animal data. If any test article treatment group yielded an SI value of 1.6 or more, then the test article was considered to be a sensitiser.

 

 

All animals survived the in-life phase of the study. All animals in the Screen and the QIT were found to be normal. There were no difficulties noted in administration of the test article or with its adherence to the dosed ears. Body weight changes were normal. Ear thickness measurements and individual animal observations indicated that all test article-treated groups in the screen had increased ear thickness (over 25%). In the Quantitative Irritation Test the 1.0% and 2.5% (w/v) test article-treated groups had increased ear thickness (over 25%). In the main test, none of the test article treated groups had ear swelling or resulted in more-than-moderate local dermal irritation. In the preliminary screen, all test article treatments resulted in no to very slight erythema. In the Quantitative Irritation Test, there was no to very slight erythema in 0.5%, 1.0%, and 2.5% (w/v) test article-treated groups, and no to very slight erythema in the 0.25% and 0.5% (w/v) test article-treated groups. The SI of the positive control group, 25% HCA, was 3.1. The group SI values for the test article were 1 (0.1% w/v test article), 1.3 (0.25% w/v test article) and 1 (0.5% w/v test article).

 

 

Thus, topical application of test article at 0.1%, 0.25%, and 0.5% (w/v) in DMSO resulted in SI values less than 1.6 (SI < 1.6), and therefore the test article is not a dermal sensitiser in the Local Lymph Node Assay in Mice.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Commercial KMPS products contain the impurity dipotassium peroxodisulphate (CAS No. 7727-21-1, EC No. 231-781-8), which has a harmonised classification according to Annex VI of the CLP Regulation as a skin sensitiser category 1 and a respiratory sensitiser category 1 (H317 / H334). Dipotassium peroxodisulphate, also known as potassium perdisulphate or potassium persulphate, is an unavoidable impurity formed during the manufacture of KMPS. The presence of such a classified impurity in a product at a concentration at or above 1.0% in the absence of evidence to the contrary causes the product itself to be classified as a skin sensitiser category 1 and a respiratory sensitiser category 1 (H317 / H334) according to the mixture rule as given in the CLP Regulation EC No 1272/2008.

 

The skin sensitisation potential of KMPS (triple salt) was originally investigated in the guinea pig maximisation test in five studies according to EU Method B.6 and/or OECD Guideline 406 and was shown to be non-sensitising. As all guinea pig maximisation tests were conducted with test materials containing 2% or unknown amounts of the potassium perdisulphate impurity, an additional sensitisation test was conducted with 2.86% impurity to ensure that also variable levels of dipotassium peroxodisulphate (concentration range: ≥0% and ≤3%, see IUCLID section 1.2) do not lead to a sensitising effects.

 

In this additional test, the skin sensitisation potential of KMPS (triple salt) was investigated in a Local Lymph Node Assay according to OECD guideline 442B and GLP utilizing the BrdU ELISA method:

 

WoE, Local Lymph Node Assay, mice, RL1, 2020

The test article was tested for solubility at 50% (w/v) in acetone:olive oil (4:1, AOO), dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), 2-butanone, and propylene glycol (PG). Test article solutions could not be made in AOO, DMF, 2-butanone, or PG. A soluble opaque off-white liquid was achieved with DMSO. Based on the solubility results, DMSO was chosen as the vehicle for the study. A preliminary dermal irritation screen was conducted with three groups (two animals per group) of healthy female CBA/J mice to determine the concentrations of the test article to be used in the main Local Lymph Node study. Three concentrations of the test article were tested: 5%, 10%, and 25% (w/v) in DMSO. The results of this preliminary screen indicated that all test article treatments resulted in dermal irritation and increased ear thickness (over 25%). Therefore, a quantitative irritation test (QIT) was performed, with six dose groups of two animals each, at test article concentrations of 0.05%, 0.1%, 0.25%, 0.5%, 1.0%, and 2.5% (w/v). The 1.0% and 2.5% (w/v) test article treatments resulted in dermal irritation and an increase in ear thickness. Test article residue at the dose site was observed in the 10% and 25% (w/v) test article-treated groups; however, this did not interfere with ear measurements. Based on these results, test article concentrations of 0.1%, 0.25%, and 0.5% (w/v) in DMSO were chosen for the main Local Lymph Node assay.

The main test was conducted with five separate groups of healthy female CBA/J mice (five animals per group). Three groups were treated with increasing concentrations of the test article. A vehicle control group was treated with DMSO and another group was treated with the positive control, alphahexylcinnamaldehyde in DMSO (25% HCA), in the exact same manner. The test article, vehicle control, and positive control solutions were administered by topical application to the dorsum of each ear, once daily for three consecutive days. All animals in the study were observed once daily throughout the study for clinical signs, either of local irritation at the application site or systemic toxicity, and for mortality. Body weights were measured on Day 1 prior to first dose and on Day 6 prior to euthanasia. The mice in the main test were given an intraperitoneal injection of the thymidine analog 5-bromo-2’-deoxy-uridine (BrdU) approximately 24 hours prior to euthanasia. After euthanasia, the auricular lymph nodes were isolated; single-cell suspensions of lymph node cells (LNC) were generated, then aliquots were permeabilized and denatured in triplicate in a 96-well microplate for analysis by ELISA for BrdU incorporation. The Stimulation Index (SI) was calculated by dividing the mean optical density of the LNC suspension for each animal by the mean optical density of the vehicle control group LNC suspension. The mean SI and standard deviation were calculated for each group from the individual animal data. If any test article treatment group yielded an SI value of 1.6 or more, then the test article was considered to be a sensitiser.

All animals survived the in-life phase of the study. All animals in the Screen and the QIT were found to be normal. There were no difficulties noted in administration of the test article or with its adherence to the dosed ears. Body weight changes were normal. Ear thickness measurements and individual animal observations indicated that all test article-treated groups in the screen had increased ear thickness (over 25%). In the Quantitative Irritation Test the 1.0% and 2.5% (w/v) test article-treated groups had increased ear thickness (over 25%). In the main test, none of the test article treated groups had ear swelling or resulted in more-than-moderate local dermal irritation. In the preliminary screen, all test article treatments resulted in no to very slight erythema. In the Quantitative Irritation Test, there was no to very slight erythema in 0.5%, 1.0%, and 2.5% (w/v) test article-treated groups, and no to very slight erythema in the 0.25% and 0.5% (w/v) test article-treated groups. The SI of the positive control group, 25% HCA, was 3.1. The group SI values for the test article were 1 (0.1% w/v test article), 1.3 (0.25% w/v test article) and 1 (0.5% w/v test article).

Thus, topical application of test article at 0.1%, 0.25%, and 0.5% (w/v) in DMSO resulted in SI values less than 1.6 (SI < 1.6), and therefore the test article is not a dermal sensitiser in the Local Lymph Node Assay in Mice.

 

WoE, Guinea Pig Maximisation Test, guinea pigs, RL1, 2001

In a (Klimisch score 1) study according to OECD guideline 406, KMPS (triple salt), containing 2% of the impurity dipotassium peroxodisulphate was applied to 10 male albino guinea pigs in the following concentrations during the study:

1. a) for the intradermal induction: FCA (Freund’s Complete Adjuvant) in water (1:1), test substance diluted to 0.25 % (w/v) with water, test substance diluted to 0.25 % (w/v) by a 1:1 mixture (v/v) of FCA and water


2. b) for topical induction: application of 20 % (w/v) test material in water


3. c) for the challenge: application of 5 and 2.5 % (w/v) test material in water each were used for the test.

5 control animals were treated Control were treated topically in the same manner than the test group animals by applying 2.5 and 5% (w/v) solution of the compound in water to the clipped flanks. Following intradermal injection necrosis was recorded at sites receiving FCA (Freund’s complete adjuvants) in test and control animals. No irritation was seen in the test animals at sites receiving KMPS (triple salt) at 0.25 % (w/v) in water for irrigation and no irritation was observed in control animals receiving water for irrigation. Following topical application of KMPS (triple salt) at 20 % (w/v) in water for irrigation, topical well defined to moderate erythema, accompanied by blanching of the dose site was observed in all test animals. No erythema was seen in the control group. Following challenge application no dermal reactions were seen in any of the test or control animals. Consequently, the test item was considered non-sensitising and no indication for skin sensitising properties observed.

 

WoE, Guinea Pig Maximisation Test, guinea pigs, RL2, 1992

A Guinea Pig Maximisation Test according to OECD guideline 406 and GLP on another KMPS product was assigned a Klimisch reliability score of 2 because the identity and purity of the test material was not fully specified in the study report. No skin sensitising properties were observed on epidermal challenge (test substance concentration 3%) after intradermal (test substance concentration 0.05%) and epidermal (test substance concentration 10%) induction exposures.

 

WoE, Guinea Pig Maximisation Test, guinea pigs, RL3, 1992

Another Guinea Pig Maximisation Test similar to OECD guideline 406 (non-GLP) was considered not reliable as the identity and purity of the test material was not specified in the study report and a high incidence of skin reactions was observed in both control and test groups. revealed an equivocal result, as positive reactions were observed in both the control group and the test substance group animals after application of distilled water only.

 

WoE, Buehler Test, guinea pigs, RL3, 1985

Two other supporting studies following the modified Buehler method, are included in the REACH dossier. These studies are not considered fully reliable because they were not performed according to GLP, which was not required when they was performed, and the identity and purity of the test materials were not specified in the study report. All results show no sensitising properties as a result of exposure to KMPS (in both studies the induction test substance concentration was 25% and the challenge test substance concentration was 6%). Hence, despite the recognised deficiencies in the reporting of these studies, the animal studies performed using recognised and robust methods show that KMPS does not have skin sensitisation properties.

 

Overall conclusion

The available animal studies for skin sensitisation show that KMPS does not have skin sensitising properties, also if maximum levels of up to 3% of the sensitising impurity peroxodisulphate are contained.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

KMPS was shown not to be a skin sensitiser in different animal studies, also if maximum levels up to 3% of the impurity dipotassium peroxodisulphate are contained.

 

In different medical health surveillance records and experience on workers no respiratory sensitisation effects have been noted (see IUCLID section 7.10).

 

Overall conclusion

KMPS is considered not to be a respiratory sensitiser in humans.

Justification for classification or non-classification

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

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. Evidence from animal studies, human testing and practical experience show that classification criteria for dermal or respiratory sensitisation are not fulfilled.

 

Based on this data, the substance is not considered to be classified for skin sensitisation under Regulation (EC) No 1272/2008, as amended for the seventeenth time in Regulation (EU) 2021/849.

 

Based on medical health surveillance records and experience on workers no respiratory sensitisation effects have been noted. The test substance is not classified regarding respiratory sensitisation according to Regulation 1272/2008/EC (CLP), as amended for the seventeenth time in Regulation (EU) 2021/849.