Iron(II) fumarate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Weight of evidence approach based on various test chemicals

Justification for type of information:

Weight of evidence approach based on various test chemicals

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data is from peer reviewed journals

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

(OECD, 1992), EC guideline B.6 (EC, 1996) and the US EPA Guideline OPPTS 870.2600 (US EPA, 2003).

Principles of method if other than guideline:

The guinea pig maximization test was performed according to OECD Test Guideline No. 406

GLP compliance:

not specified

Remarks:

data is from publication

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

not specified

Species:

guinea pig

Strain:

other: SPF-Hsd Poc: DH

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Female guinea pigs were purchased from Harlan Winkelmann (Borchen, Germany).
- Females (if applicable) nulliparous and non-pregnant: [yes/no/not specified] : yes
- Microbiological status of animals, when known:
- Age at study initiation:
- Weight at study initiation: 300–500 g at the commencement of the study
- Housing: The guinea pigs were housed in groups in Terluran cages on Altromin saw fiber bedding or Lignocel bedding
- Diet (e.g. ad libitum): Altromin 3122 maintenance diet for guinea pigs or ssniff Ms-H, 4mm V2233-000 complete diet for guinea pigs, rich in crude fiber, totally-pathogen-free), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: The acclimatization period was at least five days.
- Indication of any skin lesions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 degrees C,
- Humidity (%): 55 ± 10%
- Air changes (per hr): air change rate of at least 10 times/hour
- Photoperiod (hrs dark / hrs light): 12-h light/ dark cycle
- IN-LIFE DATES: From: To:

Route:

intradermal

Vehicle:

cotton seed oil

Concentration / amount:

Intradermal = 5% test chemical in cottonseed oil

Adequacy of induction:

not specified

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

Epicuteanous = 0.5 g of 25% test chemical in vaseline

Day(s)/duration:

48 h

Adequacy of induction:

non-irritant substance, but skin pre-treated with 10% SDS

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

0.5 g of 10% test chemical in vaseline

Day(s)/duration:

24 h

Adequacy of challenge:

not specified

No. of animals per dose:

Each test was performed with 10 test and five control animals

Details on study design:

RANGE FINDING TESTS: A range finding for the test concentrations was performed in a preliminary experiment assessing the irritant effects of the test compounds after intradermal and
topical application.

MAIN STUDY
A. INDUCTION EXPOSURE - Intradermal exposure
- No. of exposures: single
- Exposure period:
- Test groups: 10
- Control group: 5
- Site:
- Frequency of applications: three pairs of injections of 0.1 ml each of the test compound i.e 5% test chemical in cottonseed oil ; the test compound in a 1:1 (v/v) mixture of FCA/physiological saline and a 1:1 (v/v) mixture of FCA/physiological saline without test compound.
- Duration: 24 hours
- Concentrations: three pairs of injections of 0.1 ml each of the test compound i.e 5% test chemical in cottonseed oil ; the test compound in a 1:1 (v/v) mixture of FCA/physiological saline and a 1:1 (v/v) mixture of FCA/physiological saline without test compound.

A. INDUCTION EXPOSURE - Intradermal exposure
- No. of exposures: single
- Exposure period: 48 hours
- Test groups: 10
- Control group: 5
- Site:
- Frequency of applications: single
- Duration: 48 hours
- Concentrations: 0.5 g of the 25% test chemical in vaseline


B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day(s) of challenge:
- Exposure period:
- Test groups:
- Control group:
- Site:
- Concentrations:
- Evaluation (hr after challenge): 48 hours and 72 hours

OTHER:

Positive control substance(s):

not specified

Reading:

1st reading

Hours after challenge:

72

Group:

test chemical

Dose level:

10% test chemical in vaseline

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No signs of systemic toxicity were observed for the test substances in any of the animals

Remarks on result:

no indication of skin sensitisation

Interpretation of results:

other: not sensitizing

Conclusions:

No signs of systemic toxicity were observed for the test chemical in any of the animals. The test chemical caused a grade 1 skin reaction in one animal of the treatment group. The skin reaction was observed 24 and 48 h after patch removal. As from the visible inspection of the treated skin site, a sensitizing reaction cannot be differentiated from a skin irritation reaction, a re-challenge was performed in order to clarify this question.After re-challenge with the test chemical, the animals that had reacted after the fist challenge failed to react. Since the skin reaction observed after the fist challenge was not reproducible upon re-challenge the animals were not considered sensitized against the test chemical. Hence, the test chemical can be considered to be not sensitzing to skin.

Executive summary:

The guinea pig maximization test was performed according to OECD Test Guideline No. 406 (OECD, 1992), EC guideline B.6 (EC, 1996) and the US EPA Guideline OPPTS 870.2600 (US EPA, 2003). A range finding for the test concentrations was performed in a preliminary experiment assessing the irritant effects of the test compounds after intradermal and topical application. Each test was performed with 10 test and five control animals because this number of animals has been reported as sufficient for the GPMT. The animals received two induction treatments, an intradermal injection (day 0) and a topical application (day 7), and a topical challenge treatment on day 20. In addition to the requirements of the OECD guideline, a second topical challenge was performed on day 28 in all experiments. The intradermal induction comprised three pairs of injections of 0.1 ml each of 5% test chemical in cottonseed oil, the test compound in a 1:1 (v/v) mixture of FCA/physiological saline and a 1:1 (v/v) mixture of FCA/physiological saline without test compound. The control animals received applications without test substance. Approximately 24 h before the topical induction treatment (day 6) the test animals received a topical application of 0.5 ml of 10% sodium lauryl sulphate in Vaseline in order to create a mild local skin irritation. For test compounds which themselves caused a clear skin irritation at the topical induction concentration used no pretreatment with sodium lauryl sulphate was done. For the topical induction, a gauze patch with 0.5 g of the test chemical was applied to the clipped skin and held under occlusion for 48 h. Control animals received the vehicle without test compound. After a resting period, topical challenge treatments were performed on day 20 and on day 28 by applying 0.5 g of 5% test chemical in vaseline for 24 h under occlusion onto the clipped skin of both test and control animals. Approximately 21 h after removal of the challenge patch, the skin area was cleaned and cleared of hair. About 24 and 48 h after removal of the patch, the skin reaction was observed and recorded according to the Magnusson and Kligman grading scale: grade 0, no reaction, grade 1, slight edema and/or erythema; grade 2, well defined edema and/or erythema; grade 3, moderate edema and/or erythema; grade 4, severe edema and/or erythema. No signs of systemic toxicity were observed for the test chemical in any of the animals. The test chemical caused a grade 1 skin reaction in one animal of the treatment group. The skin reaction was observed 24 and 48 h after patch removal. As from the visible inspection of the treated skin site, a sensitizing reaction cannot be differentiated from a skin irritation reaction, a re-challenge was performed in order to clarify this question.After re-challenge with the test chemical, the animals that had reacted after the fist challenge failed to react. Since the skin reaction observed after the fist challenge was not reproducible upon re-challenge the animals were not considered sensitized against the test chemical. Hence, the test chemical can be considered to be not sensitzing to skin.

Cross-reference 2

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

data is from safety assessment reports

Qualifier:

according to guideline

Guideline:

other: Patch test

Principles of method if other than guideline:

Human patch test data for the test chemical

GLP compliance:

not specified

Type of study:

patch test

Justification for non-LLNA method:

not specified

Species:

other: humans

Strain:

not specified

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source:
- Females (if applicable) nulliparous and non-pregnant: [yes/no/not specified]
- Microbiological status of animals, when known:
- Age at study initiation: 44 year old male
- Weight at study initiation:
- Housing:
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period:
- Indication of any skin lesions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):
- IN-LIFE DATES: From: To:

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

saturated solution of the test chemical

Adequacy of induction:

not specified

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

saturated solution of the test chemical

Adequacy of challenge:

not specified

No. of animals per dose:

1

Details on study design:

RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period:
- Test groups:
- Control group:
- Site:
- Frequency of applications:
- Duration:
- Concentrations: saturated solution of the test chemical

B. CHALLENGE EXPOSURE
- No. of exposures: 3
- Day(s) of challenge:
- Exposure period:
- Test groups:
- Control group:
- Site:
- Concentrations: saturated solution of the test chemical
- Evaluation (hr after challenge): 48 and 72 hours

OTHER: A 44 year old male presented with eczema-type reactions on his skin in areas that were exposed to steel particles during his job using high pressure cleaning equipment to remove steel particles from construction elements. An initial patch test using steel particles resulted in erythema after 48 hours. The reaction had increased at 72 hours. Further tests were carried out. Various metals salts were tested in the patch test. These tests were repeated three times with identical results. About 30 people were used for the control test

Challenge controls:

30 people served as control

Positive control substance(s):

not specified

Reading:

1st reading

Hours after challenge:

72

Group:

test chemical

Dose level:

saturated solution of the test chemical

No. with + reactions:

0

Total no. in group:

1

Clinical observations:

no dermal reactions were observed

Remarks on result:

no indication of skin sensitisation

Interpretation of results:

other: not sensitizing

Conclusions:

The test chemical failed to produce any sensitization reaction at 48 and 72 hours, hence it can be considered to be not sensitizing to skin.

Executive summary:

A 44 year old male presented with eczema-type reactions on his skin in areas that were exposed to steel particles during his job using high pressure cleaning equipment to remove steel particles from construction elements. An initial patch test using steel particles resulted in erythema after 48 hours. The reaction had increased at 72 hours. Further tests were carried out. Various metals salts were tested in the patch test. These tests were repeated three times with identical results. About 30 people were used for the control test. A patch containing saturated solution of the test chemical was applied on the skin of the male volunteer and observed for reactions till 72 hours. The test chemical failed to produce any sensitization reaction at 48 and 72 hours, hence it can be considered to be not sensitizing to skin.

Data source

Reference

Reference Type:

review article or handbook

Title:

WoE report for the skin sensitization potential of the test chemical

Author:

Sustainability Support Services (Europe) AB

Year:

2019

Bibliographic source:

Sustainability Support Services (Europe) AB, 2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Weight of evidence approach based on various test chemicals

Principles of method if other than guideline:

Weight of evidence approach based on various test chemicals

GLP compliance:

not specified

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Iron(II) fumarate

EC Number:

205-447-7

EC Name:

Iron(II) fumarate

Cas Number:

141-01-5

Molecular formula:

C4H4O4.Fe

IUPAC Name:

iron(II) fumarate

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): Ferrous fumarate
- Molecular formula: C4H4O4.Fe
- Molecular weight : 169.901 g/mol
- Smiles notation : [Fe+2].[O-]C(\C=C\C(=O)[O-])=O
- InChl : 1S/C4H4O4.Fe/c5-3(6)1-2-4(7)8;/h1-2H,(H,5,6)(H,7,8);/q;+2/p-2/b2-1+;
- Substance type:Organic
- Physical state:Solid

In vivo test system

Test animals

Species:

other: 2. guinea pigs; 3. humans

Strain:

other: 2. SPF-Hsd Poc: DH

Sex:

female

Details on test animals and environmental conditions:

2. TEST ANIMALS
- Source: Female guinea pigs were purchased from Harlan Winkelmann (Borchen, Germany).
- Females (if applicable) nulliparous and non-pregnant: [yes/no/not specified] : yes
- Microbiological status of animals, when known:
- Age at study initiation:
- Weight at study initiation: 300–500 g at the commencement of the study
- Housing: The guinea pigs were housed in groups in Terluran cages on Altromin saw fiber bedding or Lignocel bedding
- Diet (e.g. ad libitum): Altromin 3122 maintenance diet for guinea pigs or ssniff Ms-H, 4mm V2233-000 complete diet for guinea pigs, rich in crude fiber, totally-pathogen-free), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: The acclimatization period was at least five days.
- Indication of any skin lesions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 degrees C,
- Humidity (%): 55 ± 10%
- Air changes (per hr): air change rate of at least 10 times/hour
- Photoperiod (hrs dark / hrs light): 12-h light/ dark cycle
- IN-LIFE DATES: From: To:
3.TEST ANIMALS
- Source:
- Females (if applicable) nulliparous and non-pregnant: [yes/no/not specified]
- Microbiological status of animals, when known:
- Age at study initiation: 44 year old male
- Weight at study initiation:
- Housing:
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period:
- Indication of any skin lesions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):
- IN-LIFE DATES: From: To:

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

cotton seed oil

Remarks:

Study 2

Concentration / amount:

Intradermal = 5% test chemical in cottonseed oil

Adequacy of induction:

not specified

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Remarks:

Study 2

Concentration / amount:

Epicuteanous = 0.5 g of 25% test chemical in vaseline

Day(s)/duration:

48 h

Adequacy of induction:

non-irritant substance, but skin pre-treated with 10% SDS

Induction 3

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Remarks:

Study 3

Concentration / amount:

saturated solution of the test chemical

Adequacy of induction:

not specified

Challenge

Challenge 1

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Remarks:

Study 2

Concentration / amount:

0.5 g of 10% test chemical in vaseline

Day(s)/duration:

24 h

Adequacy of challenge:

not specified

Challenge 2

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Remarks:

Study 3

Concentration / amount:

saturated solution of the test chemical

Adequacy of challenge:

not specified

No. of animals per dose:

2. Each test was performed with 10 test and five control animals
3. 1

Details on study design:

2. RANGE FINDING TESTS: A range finding for the test concentrations was performed in a preliminary experiment assessing the irritant effects of the test compounds after intradermal and
topical application.

MAIN STUDY
A. INDUCTION EXPOSURE - Intradermal exposure
- No. of exposures: single
- Exposure period:
- Test groups: 10
- Control group: 5
- Site:
- Frequency of applications: three pairs of injections of 0.1 ml each of the test compound i.e 5% test chemical in cottonseed oil ; the test compound in a 1:1 (v/v) mixture of FCA/physiological saline and a 1:1 (v/v) mixture of FCA/physiological saline without test compound.
- Duration: 24 hours
- Concentrations: three pairs of injections of 0.1 ml each of the test compound i.e 5% test chemical in cottonseed oil ; the test compound in a 1:1 (v/v) mixture of FCA/physiological saline and a 1:1 (v/v) mixture of FCA/physiological saline without test compound.

A. INDUCTION EXPOSURE - Intradermal exposure
- No. of exposures: single
- Exposure period: 48 hours
- Test groups: 10
- Control group: 5
- Site:
- Frequency of applications: single
- Duration: 48 hours
- Concentrations: 0.5 g of the 25% test chemical in vaseline


B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day(s) of challenge:
- Exposure period:
- Test groups:
- Control group:
- Site:
- Concentrations:
- Evaluation (hr after challenge): 48 hours and 72 hours

OTHER:

3. RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period:
- Test groups:
- Control group:
- Site:
- Frequency of applications:
- Duration:
- Concentrations: saturated solution of the test chemical

B. CHALLENGE EXPOSURE
- No. of exposures: 3
- Day(s) of challenge:
- Exposure period:
- Test groups:
- Control group:
- Site:
- Concentrations: saturated solution of the test chemical
- Evaluation (hr after challenge): 48 and 72 hours

OTHER: A 44 year old male presented with eczema-type reactions on his skin in areas that were exposed to steel particles during his job using high pressure cleaning equipment to remove steel particles from construction elements. An initial patch test using steel particles resulted in erythema after 48 hours. The reaction had increased at 72 hours. Further tests were carried out. Various metals salts were tested in the patch test. These tests were repeated three times with identical results. About 30 people were used for the control test

Challenge controls:

3. 30 control volunteers

Positive control substance(s):

not specified

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

72

Group:

test chemical

Dose level:

10% test chemical in vaseline

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No signs of systemic toxicity were observed for the test substances in any of the animals

Remarks on result:

no indication of skin sensitisation

Remarks:

Study 2

Results 2

Reading:

1st reading

Hours after challenge:

72

Group:

test chemical

Dose level:

saturated solution of the test chemical

No. with + reactions:

0

Total no. in group:

1

Clinical observations:

no dermal reactions were observed

Remarks on result:

no indication of skin sensitisation

Remarks:

Study 3

Applicant's summary and conclusion

Interpretation of results:

other: not sensitizing

Conclusions:

Based on the available studies and applying the weight of evidence approach, the test chemical can be considered to be not sensitizing to skin.

Executive summary:

Various studies have been reviewed to evaluate the dermal sensitization potential of the test chemical. The results are mentioned below:

The guinea pig maximization test was performed according to OECD Test Guideline No. 406 (OECD, 1992), EC guideline B.6 (EC, 1996) and the US EPA Guideline OPPTS 870.2600 (US EPA, 2003). A range finding for the test concentrations was performed in a preliminary experiment assessing the irritant effects of the test compounds after intradermal and topical application. Each test was performed with 10 test and five control animals because this number of animals has been reported as sufficient for the GPMT. The animals received two induction treatments, an intradermal injection (day 0) and a topical application (day 7), and a topical challenge treatment on day 20. In addition to the requirements of the OECD guideline, a second topical challenge was performed on day 28 in all experiments. The intradermal induction comprised three pairs of injections of 0.1 ml each of 5% test chemical in cottonseed oil, the test compound in a 1:1 (v/v) mixture of FCA/physiological saline and a 1:1 (v/v) mixture of FCA/physiological saline without test compound. The control animals received applications without test substance. Approximately 24 h before the topical induction treatment (day 6) the test animals received a topical application of 0.5 ml of 10% sodium lauryl sulphate in Vaseline in order to create a mild local skin irritation. For test compounds which themselves caused a clear skin irritation at the topical induction concentration used no pretreatment with sodium lauryl sulphate was done. For the topical induction, a gauze patch with 0.5 g of the test chemical was applied to the clipped skin and held under occlusion for 48 h. Control animals received the vehicle without test compound. After a resting period, topical challenge treatments were performed on day 20 and on day 28 by applying 0.5 g of 5% test chemical in vaseline for 24 h under occlusion onto the clipped skin of both test and control animals. Approximately 21 h after removal of the challenge patch, the skin area was cleaned and cleared of hair. About 24 and 48 h after removal of the patch, the skin reaction was observed and recorded according to the Magnusson and Kligman grading scale: grade 0, no reaction, grade 1, slight edema and/or erythema; grade 2, well defined edema and/or erythema; grade 3, moderate edema and/or erythema; grade 4, severe edema and/or erythema. No signs of systemic toxicity were observed for the test chemical in any of the animals. The test chemical caused a grade 1 skin reaction in one animal of the treatment group. The skin reaction was observed 24 and 48 h after patch removal. As from the visible inspection of the treated skin site, a sensitizing reaction cannot be differentiated from a skin irritation reaction, a re-challenge was performed in order to clarify this question.After re-challenge with the test chemical, the animals that had reacted after the fist challenge failed to react. Since the skin reaction observed after the fist challenge was not reproducible upon re-challenge the animals were not considered sensitized against the test chemical. Hence, the test chemical can be considered to be not sensitzing to skin.

This is supported a case study in which a 44 year old male presented with eczema-type reactions on his skin in areas that were exposed to steel particles during his job using high pressure cleaning equipment to remove steel particles from construction elements. An initial patch test using steel particles resulted in erythema after 48 hours. The reaction had increased at 72 hours. Further tests were carried out. Various metals salts were tested in the patch test. These tests were repeated three times with identical results. About 30 people were used for the control test. A patch containing saturated solution of the test chemical was applied on the skin of the male volunteer and observed for reactions till 72 hours. The test chemical failed to produce any sensitization reaction at 48 and 72 hours, hence it can be considered to be not sensitizing to skin.

Based on the available studies and applying the weight of evidence approach, the test chemical can be considered to be not sensitizing to skin.