Trisodium hydrogen ethylenediaminetetraacetate

Endpoint summary

Description of key information

skin irritation/corrosion (OECD 404): not irritating (read-across from disodium dihydrogen EDTA and tetrasodium EDTA)

eye irritation (equivalent or similar to OECD 405): irreversible eye damage (read-across from tetrasodium EDTA)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Refer to analogue justification provided in IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0.4

Max. score:

4

Remarks on result:

other:

Remarks:

source: CAS 64-02-8, BASF SE, 1982, rabbit

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days

Remarks on result:

other:

Remarks:

source: CAS 64-02-8, BASF SE, 1982, rabbit

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other:

Remarks:

source: CAS 64-02-8, BASF SE, 1982, rabbit

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other:

Remarks:

source: CAS 64-02-8, BASF SE, 1982, rabbit

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other:

Remarks:

source: CAS 64-02-8, BASF SE, 1982, rabbit

In a supporting skin irritation study performed with the source substance disodium dihydrogen EDTA (CAS 139-33-3), two rabbits were exposed occlusively for 1, 5 or 15 minutes on the back to a 50% aqueous solution of the test item (BASF SE 1973). Two additional rabbits were exposed to the same solution for 20 h on the back and at the ear. The skin of the animals was observed for 7 days. Both animals developed a mild erythema 24 h after application (grad 1 or grade 2 respectively) which was fully reversible within 48 or 72 h. No edema was observed. A well-defined erythema was observed on the ear of both animals, which was fully reversible within 48 h.

Interpretation of results:

other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

An in vivo skin irritation study in rabbits with the source substance tetrasodium EDTA (CAS 64-02-8) was selected as key result for reasons of structural similarity and data reliability. Supporting in vivo skin irritation data is given for the source substance disodium dihydrogen EDTA (CAS 139-33-3), where only mild erythema, not leading to classification, were found.
The read across approach is justified in the analogue justification. The target and source substances are considered unlikely to differ in their skin irritation potential. No in vivo skin irritation potential was found in rabbit studies for the two source substances with structural similarity. Therefore, no skin irritation potential is expected for target substance trisodium hydrogen EDTA (CAS 150-38-9).

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Refer to analogue justification provided in IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 8 days

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.5

Max. score:

4

Reversibility:

not fully reversible within: 8 days

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

3

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

not fully reversible within: 8 days

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

not fully reversible within: 8 days

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.8

Max. score:

4

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: source: CAS 64-02-8, BASF AG, 1978, rabbit

In a WoE eye irritation test, performed with source substance tetrasodium EDTA (CAS 64-02-8), instillation of 50 mg of of the test item to the eye of two rabbits resulted in redness (score 1.75), edema (score 1.25) and opacity (score 1.3) and pus formation as average 24/48/72 h after application (BASF, 1970). After 8 days a mild opacity was seen.

Tetrasodium dihydrogen EDTA has received the harmonized CLP classification Eye Dam. 1 (H318).

In a WoE eye irritation test performed with the source substance disodium dihydrogen EDTA (CAS 139-33-3), 50 mg of solid test item were instilled into the conjunctival sac of one eye of two animals. The other eye served as control and was treated with 50 mg talcum powder. The test item was not washed out and the animals were observed for 8 days. A slight redness of the eye (score 1) was observed in both animals, which was fully reversible within 8 days.

Interpretation of results:

other: CLP/GHS criteria met; classification as Eye Dam. 1, H318 required according to Regulation (EC) No. 1272/2008

Conclusions:

An in vivo eye irritation study in rabbits with the source substance tetrasodium EDTA (CAS 64-02-8) was selected for classification for reasons of structural similarity, data reliability and for reasons of a conservative (worst case) hazard assessment approach.
The read across approach is justified in the analogue justification. The target and source substances are considered unlikely to differ in their eye irritation potential. An in vivo eye damage potential was found in the rabbit study for the source substance tetrasodium dihydrogen EDTA (CAS 64-02-8) with structural similarity. For this substance there exists a CLP harmonized classification as Eye Dam. 1. Therefore, eye damaging potential is expected for the target substance trisodium hydrogen EDTA (CAS 150-38-9).

CLP: Eye Dam. 1, H318

In a WoE eye irritation test, performed with source substance tetrasodium EDTA (CAS 64-02-8), instillation of 50 mg of of the test item to the eye of two rabbits resulted in redness (score 1.75), edema (score 1.25) and opacity (score 1.3) and pus formation as average 24/48/72 h after application (BASF, 1970). After 8 days still a mild opacity was seen.

In a WoE eye irritation test performed with the source substance disodium dihydrogen EDTA (CAS 139-33-3), Only mild signs of eye irritation not requiring classification according to Regulation (EC) No. 1272/2008 were noted. The read across approach is justified in the analogue justification. However, eye damaging potential was found in in vivo studies with the surrogate substance tetrasodium dihydrogen EDTA (CAS 64-02-8). For reasons of a conservative (worst case) assessment approach data from tetrasodium EDTA were used for risk assessment and classification.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Dose not exactly specified (OECD: volume of 0.1 mL); 2 animals are used (OECD: 3); examination using fluorescein (OECD: no use of fluorescein)

Principles of method if other than guideline:

BASF-TEST: Before OECD guideline 405 was established, eye irritation was tested using an internal method. White Vienna rabbits were used. Usually approximately 50 mg of the test substance were applied to the conjunctival sac of one eye of 2 animals. In studies with solids, talcum powder was used as control. The animals were observed several times on the treatment day and up to 8 days afterwards, and findings were recorded on working days.
The eyes were not washed out after 24 h as specified in OECD Guideline 405, which could be critical for solids which thus remained on the cornea for several days, causing mechanical damage. Probably, this would have been less severe if it had been washed out after 1 day.

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 3.39; 3.15 kg

Vehicle:

unchanged (no vehicle)

Controls:

other: talcum treated eye of the same animal

Amount / concentration applied:

- Amount applied: a sharp spoon (probably approximately 50 mg)

Duration of treatment / exposure:

- substance was not washed out

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no washing was done

READING TIME POINTS
10 min, 1 h, 3 h, 24 h, 48 h, 72 h, 6 days and 8 days after application; however the data on the 48 h, 72 h and 6 days reading of animal 2 are not reported.

READING TIME POINTS USED FOR ASSESSMENT
24, 48 and 72 h for animal 1; 24 h for animal 2

SCORING SYSTEM:
For the evaluation the BASF scores given in the raw data were converted to Draize scores

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24 - 48 - 72 h for animal 1; 24 h for animal 2

Score:

0.5

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Both animals showed some redness of the conjunctivae (score 1), 24 h after application of the test susbtance. The same effect was observed in the talcum powder treated eye. This effect was fully reversible 48 h in animal 1 and at least within 8 days in animal 2.

Interpretation of results:

other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

Only mild signs of eye irritation not requiring classification according to Regulation (EC) No. 1272/2008 were noted in this WoE study with the source substance disodium dihydrogen EDTA (CAS 139-33-3). The read across approach is justified in the analogue justification.
CLP: not classified

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

yes

Remarks:

dose 50 mg (OECD: volume of 100 mg); 2 animals are used (OECD:3); no washing done (OECD: washing after 1 h), observation period 8 days (OECD: 21)

Principles of method if other than guideline:

BASF-TEST: Before OECD guideline 405 was established, eye irritation was tested using an internal method. White Vienna rabbits were used. Usually approximately 50 mg of the test substance were applied to the conjunctival sac of one eye of 2 animals. In studies with solids, talcum powder was used as control. The animals were observed several times on the treatment day and up to 8 days afterwards, and findings were recorded on working days.
The eyes were not washed out after 24 h as specified in OECD Guideline 405, which could be critical for solids which thus remained on the cornea for several days, causing mechanical damage. Probably, this would have been less severe if it had been washed out after 1 day

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Vehicle:

unchanged (no vehicle)

Controls:

other: talcum powder treated eye of the same animal

Amount / concentration applied:

50 mg

Duration of treatment / exposure:

- unspecified; substance was not washed out

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no washing was done

READING TIME POINTS
10 min, 1 h, 3 h, 24 h, 48 h, 4 d, 7d 8 days after application

READING TIME POINTS USED FOR ASSESSMENT
24 hand 48 h

SCORING SYSTEM:
For the evaluation the BASF scores given in the raw data were converted to Draize scores

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1.25

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 8 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.5

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

1.75

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.5

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

1.25

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritant / corrosive response data:

- Both animals had effects on the cornea, conjunctivae and showed chemiosis.
- Additionally there was a suppuration of the treated eye in animal 2. This effect was reversible within 8 days.
- The talcum treated eye of animal 2 showed conjunctivae score 1 after 24 h, which was fully reversible within 48 h. In animal 1 no effects on the talcum treated eye were observed 24 h after application.

Other effects:

- on day 7 animal 2 suffered from diarrhea did not eat. These symptoms disappeared on day 8.

Interpretation of results:

other: CLP/GHS criteria met; classification required according to Regulation (EC) No. 1272/2008

Conclusions:

In this eye irritation test, performed with source substance tetrasodium dihydrogen EDTA (CAS 64-02-8), instillation of 50 mg of of the test item to the eye of two rabbits resulted in redness (score 1.75), edema (score 1.25) and opacity (score 1.3) and pus formation as average 24/48/72 h after application. After 8 days still a mild opacity was seen.

For this substance there exists a CLP harmonized classification as Eye Dam. 1.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for read-across

There are no experimental data available regarding the skin and eye irritation potential of trisodium hydrogen EDTA (CAS 150-38-9). Thus, read-across from appropriate source substances tetrasodium EDTA (CAS 64-02-8) and disodium dihydrogen EDTA (CAS 139-33-3) is conducted in accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5 in order to fulfil the standard information requirements defined in Regulation (EC) No 1907/2006, Annex VII, 8.1. (Bio)transformation to common compounds of the source and target substance are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

Skin irritation

In a key study, which was performed with the source substance tetrasodium EDTA (CAS 64-02-8), according to OECD 404 the skin of 2 male and 1 female rabbits was exposed to 0.5 g of the test item for an occlusive exposure period of 4 hours. For erythema medium scores after 24/48/72 hours of 1/0/0.3 were documented, redness had resolved after 8 days. No edema was noted (BASF, 1982).

In a supporting skin irritation study performed with the source substance disodium dihydrogen EDTA (CAS 139-33-3), two rabbits were exposed occlusively for 1, 5 or 15 minutes on the back to a 50% aqueous solution of the test item (BASF 1973). Two additional rabbits were exposed to the same solution for 20 h on the back and at the ear. The skin of the animals was observed for 7 days. Both animals developed a mild erythema 24 h after application (grad 1 or grade 2 respectively) which was fully reversible within 48 or 72 h. No edema was observed. A well-defined erythema was observed on the ear of both animals, which was fully reversible within 48 h.

Eye irritation

In a WoE study which was performed with the source substance tetrasodium EDTA (CAS 64-02-8), approximately 50 mg of the test item were applied to the conjunctival sac of one eye of 2 animals. 24 to 72 h after application a mild redness (score 1), mild edema (score 0.8) and mild opacity (score 1.3) was present and after 8 days mild redness and mild opacity persisted (BASF, 1978). During all these observation times a grease-like layer was observed.

In a WoE eye irritation test, performed with source substance tetrasodium EDTA (CAS 64-02-8), instillation of 50 mg of the test item to the eye of two rabbits resulted in redness (score 1.75), edema (score 1.25) and opacity (score 1.3) and pus formation as average 24/48/72 h after application (BASF, 1970). After 8 days a mild opacity was seen.

Tetrasodium EDTA has received the harmonized CLP classification Eye Dam. 1 (H318).

In a WoE eye irritation test, performed with the source substance disodium dihydrogen EDTA (CAS 139-33-3), 50 mg of solid test item were instilled into the conjunctival sac of one eye of two animals. The other eye served as control and was treated with 50 mg talcum powder. The test item was not washed out and the animals were observed for 8 days. A slight redness of the eye (score 1) was observed in both animals, which was fully reversible within 8 days.

While both studies conducted with the source substance tetasodium EDTA (CAS 64-08-2) demonstrated eye irritating or even eye damaging effects, the study performed with the source substance disodium dihydrogen EDTA (CAS 139-33-3) resulted in only very mild eye irritation. An explanation for the inconsistent results lies in the different pH values of the surrogate substances: disodium dihydrogen EDTA has a pH ranging from 4.0 to 5.5, while tetrasodium dihydrogen EDTA has a pH value ranging from 10.7 to 11.7. The pH value of trisodium hydrogen EDTA is 7, i.e. located between the pHs of the surrogate substances. Following a conservative approach, the severe eye effects caused by tetrasodium EDTA are used for risk assessment and CLP classification purposes and the target substance trisodium hydrogen EDTA is classified as Eye Dam 1 (H318).

Justification for classification or non-classification

Based on read-across, the available data on skin irritation/corrosion do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 and are therefore conclusive but not sufficient for classification.

Based on read-across, the available data on eye irritation meet the criteria for classification according to Regulation (EC) No. 1272/2008 as Eye Dam. 1 (H318).