Alkaline earth metal and zinc hydroxy(substituted)alkanoate and oxoalkanoate

Endpoint summary

Description of key information

Skin corrosion

Read-across: WoE, TP 1646, reconstructed human epidermis (RhE) in vitro model EPISKIN™, OECD 431, GLP, negative

Skin irritation

Read-across: WoE, TP 1646, reconstructed human epidermis (RhE) in vitro model EPISKIN™, OECD 439, GLP, negative

Read-across: WoE, FF6, rabbit in vivo, OECD 405, GLP, negative

Eye irritation:

Read-across: WoE, TP 1646, bovine cornea ex vivo, OECD 437, GLP, negative

Read-across: WoE, FF6, rabbit in vivo, OECD 405, GLP, negative

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reason / purpose for cross-reference:

read-across source

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min

Value:

40

Negative controls validity:

valid

Positive controls validity:

not examined

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min

Value:

79

Negative controls validity:

valid

Positive controls validity:

not examined

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

240min

Value:

75

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

In the Main Assay, the negative controls gave the expected baseline value (Optical Density values ≥ 0.6 and ≤ 1.5) and variability (difference of viability between the two replicates lower than 30%), at each treatment time, in agreement with the guideline indications. For each treatment time, the concurrent negative control mean value is considered the baseline value of the treatment series and thus represents 100% of cell viability. The positive control caused the expected cell death (1% of cell viability, when compared to the negative control). Based on the stated criteria, the assay was regarded as valid.

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

The mean cell viability of the test item treated tissues, after the blank subtraction, was higher than 35 % at all treatment times. Based on the results obtained, the test item is identified as non-corrosive to the skin.

Executive summary:

The potential of the test item to be corrosive to the skin was investigated through an in vitro skin corrosion study, using a commercial reconstructed human epidermis (RhE) model named EPISKIN™. The experimental procedures are based on the OECD Guideline for testing of chemicals no. 431 and was performed under GLP conditions. The test item, as well as controls, were tested for their ability to impair cell viability after an exposure period of 3, 60 and 240 minutes. The final endpoint of the assay is the colorimetric measurement of MTT reduction (blue formazan salt) in the test system, being this reaction an index of cell viability. The test item was tested as supplied by the Sponsor.

A preliminary test was carried out to evaluate the compatibility of the test item with the test system. In a first step, the test item was assayed for the ability of reducing MTT per se. After addition of the test item to the MTT solution, a blue colour was noted indicating that the test item could direct interact with MTT. In a second step, the test item was assayed for the ability of colouring water per se. A colourless solution was observed, indicating that the test item has no potential interfering ability. Based on these results, an additional control for non specific MTT reduction (NSMTT) was added in the Main Assay.

In the Main Assay, for each treatment time, the test item (physical state: solid) was applied as supplied in two replicates, at the treatment level of 20 ± 2 mg/epidermis unit, each measuring 0.38 cm² (treatment level: 52.6 mg/cm²). Positive and negative controls (glacial acetic acid and physiological saline, respectively) were concurrently tested, in the same number of replicates and test conditions at the treatment level of 50 μL/epidermis unit. Positive control was included only at the longest treatment time of 240 minutes, while a negative control was included for each treatment time.

In the Main Assay, the negative controls gave the expected baseline value (Optical Density values ≥ 0.6 and ≤ 1.5) and variability (difference of viability between the two replicates lower than 30%), at each treatment time, in agreement with the guideline indications. For each treatment time, the concurrent negative control mean value is considered the baseline value of the treatment series and thus represents 100% of cell viability.

The positive control caused the expected cell death (1% of cell viability, when compared to the negative control).

Based on the stated criteria, the assay was regarded as valid.

The NSMTT values were higher than 5%, but lower than 50% at all treatment times, thus these values were subtracted from the concurrent mean OD value of the test item treated tissues, to evaluate the actual viability.

The mean cell viability of the test item treated tissues, after the blank subtraction, was higher than 35% at all treatment times. Based on the results obtained, the test item is identified as non-corrosive to the skin.

Due to the similar composition this result is also valid for the target substance TP 1740.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reason / purpose for cross-reference:

read-across source

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

89

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The mean Optical Density of Blank Controls was 0.031, lower than the maximum acceptable value (0.1). The negative control gave the expected baseline value (Optical Density values of the three replicates higher than 0.6) and variability (Standard Deviation of the % viability lower or equal to 18), in agreement with guideline indications. According to the method, the mean value is considered the baseline value of the experiment and thus represents 100% of cell viability. Positive control results indicated an appropriate cell death with an acceptable relative cell viability (5% of the negative control value). Variability between replicates gave also the expected value (SD of %viability = 0.8). Based on the stated criteria, mean viability, expressed as percentage of the negative control, lower or equal to 40% and standard deviation of % viability equal or lower than 18, the study was accepted as valid. The NSMTT value was -1%. Based on this result, only the OD-blank background subtraction was performed and the mean cell viability was 89%, when compared to the negative control. Acceptable intra-replicate variability was obtained (SD of % viability = 2.6 lower than 18).

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

The mean cell viability of the test item treated tissues, after the blank subtraction, was 89%.
Based on the results obtained, the test item is classified as non-irritant to the skin (UN GHS No Category).

Executive summary:

The potential of the test item to be irritant to the skin was investigated through an in vitro skin irritation study using a commercial reconstructed human epidermis (RhE) model named EPISKIN™. The experimental procedures are based on the OECD Guideline for testing of chemicals no. 439 and was performed under GLP conditions. The test item, as well as controls, were tested for their ability to impair cell viability after an exposure period of 15 minutes followed by a 42 ± 1 hour recovery period. The final endpoint of the assay is the colorimetric measurement of MTT reduction (blue formazan salt) in the test system being this reaction an index of cell viability. The test item was tested as supplied by the Sponsor.

Before the Main Assay, a preliminary test was carried out to evaluate the compatibility of the test item with the test system. In a first step, the test item was assayed for the ability of reducing MTT per se. A blue solution was noted at the end of the incubation period, indicating that the test item could direct interact with MTT. In a second step, the test item was assayed for the ability of colouring water per se. A colourless solution was observed, indicating that the test item has no potential colour interfering ability. Based on these results, an additional control for non specific MTT reduction (NSMTT) was added in the Main Assay.

In the Main Assay, the test item was applied as supplied in three replicates at the treatment level of 20±2 mg/epidermis unit, each measuring 0.38 cm² (treatment level: 53 μL/cm²). Positive and negative controls [a 5% (w/v) sodium dodecyl sulphate solution in water and Dulbecco’s phosphate buffered saline (D-PBS), respectively] were concurrently tested, in the same number of replicates and test conditions at the treatment level of 20 μL/epidermis unit. Non specific MTT reduction (NSMTT) was evaluated using two killed tissues and compared with negative control performed with alive tissues.

In the Main Assay, the negative control gave the expected baseline value (Optical Density values of the three replicates higher than 0.6) and variability [Standard Deviation (SD) of % viability lower or equal to 18], in agreement with the guideline indications. According to the method, the negative control mean value is considered the baseline value of the experiment and thus represents 100% of cell viability.

The positive control caused the expected cell death (5% of cell viability when compared to the negative control) and variability (SD of % viability equal to 0.8). Based on the stated criteria (mean viability ≤ 40% and SD of % viability ≤ 18), the assay was regarded as valid.

The NSMTT value was −1%, thus only the OD-blank background subtraction was performed. The test item did not induce cell death in any replicate, the mean cell viability after the blank subtraction was 89% when compared to the negative control. Intra-replicate variability was acceptable with a SD of % viability value equal to 2.6 (lower than 18, as stated in the Study Protocol).

Based on the results obtained, the source substance is classified as non-irritant to the skin (UN GHS No Category). Due to the similar composition this result is also valid for the target substance TP 1740.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

1

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

1

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

1

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

1

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

1

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

1

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The treated skin areas showed only a slight erythema one hour after patch removal in two animals. Other signs of irritation were not observed. All effects were fully reversible within 24 hours.

Other effects:

No animal died in the course of the investigation, clinical signs were not observed. A necropsy was not made, because no clinical signs were observed.
The control skin areas of the animals did not show any alterations at any observation time.
No systemic toxic effects were observed.

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

The test item is not irritant to the skin

Executive summary:

The acute dermal irritation/corrosion of the test item was conducted following OECD 405 and under GLP compliance. On three albino rabbits the test article was applied in a single dose of 0.5 g to a shaved dorsal area of trunk and covered with agauze patch and aluminium foil which was held in contact with the skin by an occlusive dressing. Exposure duration was 4 hours. Thereafter residual substance was removed using water.

Animals were examined for mortality, clinical signs and signs of irritation response 60 minutes, 24, 48, 72 hours after patch removal.

Only a slight erythema was observed one hour after patch removal in two animals. No animal died or showed clinical signs in the course of testing. According to the results of this testing the test item is not classified.

The source substance contains the major organic moiety (hydroxysulfonatoactetate) that is identical to the target substance. Therefore, the target substance TP 1740 is also considered to be not irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

2.78

Vehicle controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

2

Value:

0.56

Vehicle controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

3

Value:

0.97

Vehicle controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

The test item showed no effects on the cornea of the bovine eye. The calculated IVIS is 1.44.

Executive summary:

This in vitro study according to OECD guideline 437 and EU Method B.47 under GLP-conditions, was performed to assess corneal damage potential of the test item by quantitative measurements of changes in opacity and permeability in a bovine cornea.

Two experiments were performed. The first experiment was considered invalid, because all of the three replicates caused different classifications. The first experiment is not reported but the raw data are kept in the GLP-archive of the test facility. The second experiment was considered as valid. The test item was brought onto the cornea of a bovine eye, which previously had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been determined. The test item was incubated on the cornea for 4 hours at 32 ± 1°C. After removal of the test item, opacity and permeability values were measured. The test item was tested as 20% solution in HBSS. Under the conditions of this test, the test item showed no effects on the cornea of the bovine eye. The calculated IVIS (In Vitro Irritancy Score) is 1.44. According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage. The negative control (HBSS) and the positive control (20% imidazole solution) have met the validity criteria. No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid. Therefore, the test item TP 1646 is not classified for eye irritation. Due to the similar composition this result is also valid for the target substance TP 1740.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Remarks:

up to day 7

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Remarks:

up to day 7

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Remarks:

up to day 7

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Remarks:

up to day 7

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Remarks:

up to day 7

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Remarks:

up to day 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 1 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

48 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

48 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

48 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 1 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 1 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

Any observed effect was fully reversible within 7 days after exposure.

Other effects:

No animal died during the course of the investigation. Vocalisation was noticed briefly after instillation of the test article in animal No. 2. Necropsy was not carried out because no clinical signs were observed at the end of the observation period.
In the treated eye a briefly serous lacrimation was observed in all animals one hour after instillation. The control eyes of the animals showed no alterations at any observation time.
The Cornea of the treated eye showed a slight opacity in one animal only one hour after instillation. The iris was not affected.

The conjunctivae of the treated eyes were slightly red in one animal and clearly red in two animals one hour after instillation and clearly red 24 hours after instillation in all three animals. This clear redness continued in all animals until 48 hours after instillation. Then the redness faded in all animals and a redness was not more observed on day 7. One hour after instillation a clear swelling of conjunctivae was observed in two animals and a swelling with lids about half closed was observed in one animal. This swelling faded quickly and 24 hours (in two animals) or 48 hours (in one animal) after instillation a swelling was not more observed. 7 days after instillation no signs of irritation were observed.

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

The test item is non-irritant to the eye.

Executive summary:

Determination of the acute eye irritation/corrosion of the test item was conducted following OECD guideline 405 under GLP compliance. On three albino rabbits the test article was applied at a single dose of 0.1 g to one of the eyes in each animal. The eyes were washed out 24 hours after instillation of the test article using distilled water. The untreated eye was used for control. The animals were examined for clinical signs and the eyes were examined for lesions of the conjunctivae, cornea and iris 60 minutes, 24, 48 and 72 hours and from day 4 until day 7 after instillation of the test article. The instillation of the test article caused a clear redness for 48 hours and a briefly clear swelling of the conjunctivae in all 3 animals. The cornea was only slightly affected and the iris was not affected. All observed effects were fully reversible within 7 days after exposure. No signs of systemic effects of the test article were observed in the course of testing. Thus, the test item is not classified for eye irritation.

The source substance contains a major organic moiety (hydroxysulfonatoacetates) that is identical to the target substance. Therefore, this study shows that no eye irritating effects can be attributed to the hydroxysulfonatoacetate moiety of the target substance TP 1740.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin corrosion

The potential of the test item to be corrosive to the skin was investigated through anin vitroskin corrosion study, using a commercial reconstructed human epidermis (RhE) model named EPISKIN™. The experimental procedures are based on the OECD Guideline for testing of chemicals no. 431 and was performed under GLP conditions. The test item, as well as controls, were tested for their ability to impair cell viability after an exposure period of 3, 60 and 240 minutes. The final endpoint of the assay is the colorimetric measurement of MTT reduction (blue formazan salt) in the test system, being this reaction an index of cell viability. The test item was tested as supplied by the Sponsor.

A preliminary test was carried out to evaluate the compatibility of the test item with the test system. In a first step, the test item was assayed for the ability of reducing MTT per se. After addition of the test item to the MTT solution, a blue colour was noted indicating that the test item could direct interact with MTT. In a second step, the test item was assayed for the ability of colouring water per se. A colourless solution was observed, indicating that the test item has no potential interfering ability. Based on these results, an additional control for non specific MTT reduction (NSMTT) was added in the Main Assay.

In the Main Assay, for each treatment time, the test item (physical state: solid) was applied as supplied in two replicates, at the treatment level of 20 ± 2 mg/epidermis unit, each measuring 0.38 cm²(treatment level: 52.6 mg/cm²). Positive and negative controls (glacial acetic acid and physiological saline, respectively) were concurrently tested, in the same number of replicates and test conditions at the treatment level of 50 μL/epidermis unit. Positive control was included only at the longest treatment time of 240 minutes, while a negative control was included for each treatment time.

In the Main Assay, the negative controls gave the expected baseline value (Optical Density values ≥ 0.6 and ≤ 1.5) and variability (difference of viability between the two replicates lower than 30%), at each treatment time, in agreement with the guideline indications. For each treatment time, the concurrent negative control mean value is considered the baseline value of the treatment series and thus represents 100% of cell viability.

The positive control caused the expected cell death (1% of cell viability, when compared to the negative control).

Based on the stated criteria, the assay was regarded as valid.

The NSMTT values were higher than 5%, but lower than 50% at all treatment times, thus these values were subtracted from the concurrent mean OD value of the test item treated tissues, to evaluate the actual viability.

The mean cell viability of the test item treated tissues, after the blank subtraction, was higher than 35% at all treatment times. Based on the results obtained, the test item is identified as non-corrosive to the skin.

Due to the similar composition this result is also valid for the target substance TP 1740.

Skin irritation

Bisini, 2018b

The potential of the test item to be irritant to the skin was investigated through anin vitroskin irritation study using a commercial reconstructed human epidermis (RhE) model named EPISKIN™. The experimental procedures are based on the OECD Guideline for testing of chemicals no. 439 and was performed under GLP conditions. The test item, as well as controls, were tested for their ability to impair cell viability after an exposure period of 15 minutes followed by a 42 ± 1 hour recovery period. The final endpoint of the assay is the colorimetric measurement of MTT reduction (blue formazan salt) in the test system being this reaction an index of cell viability. The test item was tested as supplied by the Sponsor.

Before the Main Assay, a preliminary test was carried out to evaluate the compatibility of the test item with the test system. In a first step, the test item was assayed for the ability of reducing MTT per se. A blue solution was noted at the end of the incubation period, indicating that the test item could direct interact with MTT. In a second step, the test item was assayed for the ability of colouring water per se. A colourless solution was observed, indicating that the test item has no potential colour interfering ability. Based on these results, an additional control for non specific MTT reduction (NSMTT) was added in the Main Assay.

In the Main Assay, the test item was applied as supplied in three replicates at the treatment level of 20±2 mg/epidermis unit, each measuring 0.38 cm²(treatment level: 53 μL/cm²). Positive and negative controls [a 5% (w/v) sodium dodecyl sulphate solution in water and Dulbecco’s phosphate buffered saline (D-PBS), respectively] were concurrently tested, in the same number of replicates and test conditions at the treatment level of 20 μL/epidermis unit. Non specific MTT reduction (NSMTT) was evaluated using two killed tissues and compared with negative control performed with alive tissues.

In the Main Assay, the negative control gave the expected baseline value (Optical Density values of the three replicates higher than 0.6) and variability [Standard Deviation (SD) of % viability lower or equal to 18], in agreement with the guideline indications. According to the method, the negative control mean value is considered the baseline value of the experiment and thus represents 100% of cell viability.

The positive control caused the expected cell death (5% of cell viability when compared to the negative control) and variability (SD of % viability equal to 0.8). Based on the stated criteria (mean viability ≤ 40% and SD of % viability ≤ 18), the assay was regarded as valid.

The NSMTT value was −1%, thus only the OD-blank background subtraction was performed. The test item did not induce cell death in any replicate, the mean cell viability after the blank subtraction was 89% when compared to the negative control. Intra-replicate variability was acceptable with a SD of % viability value equal to 2.6 (lower than 18, as stated in the Study Protocol).

Due to the similar composition this result is also valid for the target substance TP 1740.

Müller, 1998

The acute dermal irritation/corrosion of the test item was conducted following OECD 405 and under GLP compliance. On three albino rabbits the test article was applied in a single dose of 0.5 g to a shaved dorsal area of trunk and covered with agauze patch and aluminium foil which was held in contact with the skin by an occlusive dressing. Exposure duration was 4 hours. Thereafter residual substance was removed using water.

Animals were examined for mortality, clinical signs and signs of irritation response 60 minutes, 24, 48, 72 hours after patch removal.

Only a slight erythema was observed one hour after patch removal in two animals. No animal died or showed clinical signs in the course of testing. According to the results of this testing the test item is to classify as non-irritant to the skin.

The source substance contains the major organic moiety (hydroxysulfonatoactetate) that is identical to the target substance. Based on the results obtained, the source substance is classified as non-irritant to the skin (UN GHS No Category), which can be attributed to the hydroxysulfonatoactetate moiety of the target substance TP 1740.

Eye irritation

Andres, 2018

This in vitro study, according to OECD guideline 437 and EU Method B.47, under GLP-conditions, was performed to assess corneal damage potential of the test item by quantitative measurements of changes in opacity and permeability in a bovine cornea.

Two experiments were performed. The first experiment was considered invalid because all of the three replicates caused different classifications. The first experiment is not reported but the raw data are kept in the GLP-archive of the test facility. The second experiment was considered as valid. Only the second experiment is reported here. The test item was brought onto the cornea of a bovine eye which previously had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been determined. The test item was incubated on the cornea for 4 hours at 32 ± 1°C. After removal of the test item, opacity and permeability values were measured. The test item was tested as 20% solution in HBSS. Under the conditions of this test, the test item showed no effects on the cornea of the bovine eye. The calculated IVIS (In Vitro Irritancy Score) is 1.44. According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage. The negative control (HBSS) and the positive control (20 % imidazole solution) have met the validity criteria. No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.

Due to the similar composition this result is also valid for the target substance TP 1740.

Müller, 1998

Determination of the acute eye irritation/corrosion of the test item was conducted following OECD guideline 405 under GLP compliance. On three albino rabbits the test article was applied at a single dose of 0.1 g to one of the eyes in each animal. The eyes were washed out 24 hours after instillation of the test article using distilled water. The untreated eye was used for control. The animals were examined for clinical signs and the eyes were examined for lesions of the conjunctivae, cornea and iris 60 minutes, 24, 48 and 72 hoursand from day 4 until day 7 after instillation of the test article.

The instillation of the test article caused a clear redness for 48 hours and a briefly clear swelling of the conjunctivae in all 3 animals.

The cornea was only slight affected and the iris was not affected. All observed effects were fully reversible within 7 days after exposure.

No signs of systemic effects of the test article were observed in the course of testing. Thus, the test item is classified as non-irritant to the eye.

Due to the similar composition this result is also valid for the target substance TP 1740.

Justification for classification or non-classification

Based on the reliable test results of the source substances the target substance is not classified according to Regulation (EC) No 1272/2008.