2-Thiophenecarboxylic acid, 5-chloro-

Endpoint summary

Description of key information

Study conducted according to OECD test guideline 404; young adult New Zealand White rabbits (3/treatment) were dermally exposed to 0.5 g/patch of Clorthiphen-2-carbonsäure (100 % a.i.) for 4 hours; result: no irritation

Study conducted according to OECD test guideline 431; 5-Chlorthiophen-2-carbonsäure (100% a.i.) was applied to the three-dimensional human epidermis model tissue for an exposure period of 3 and 60 minutes in triplicates, result: Not corrosive

Study conducted according to OECD test guideline 405; 0.1 g 5-Chlorthiophen-2-carbonsäure (100 % a.i.) were instilled into the conjunctival sac of one eye of 3 young adult New Zealand White rabbits for 24 hours, result: severe irritation: Category 1

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-10-28 to 2005-12-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

22. April 2002

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

Esd:NZW

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories France, 69592 L’Arbresle, France
- Age at study initiation: young adult animals
- Weight at study initiation: 2.7 kg — 3.1 kg
- Housing: The animals were housed individually in cage units Metall/Noryl by EBECO. Excrement trays below the cages contained low dust wood granulate bedding (J. Rettenmaier & Söhne, 73494 Rosenberg, Germany).
- Diet (e.g. ad libitum): The animals received the standard diet “SsniffK-Z” 4mm (manufacturer: Ssniff Spezialdiäten GmbH, 59494 Soest, Germany), approximately 100 g per animal per day
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3
- Humidity (%): 50±25
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Remarks:

pulverizede test item moistened with water

Controls:

other:

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 g

Duration of treatment / exposure:

4 h

Observation period:

72 h

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: 0.5 g of the pulverized test substance moistened with Aqua p.i. (to ensure good contact with the skin) was applied to the skin of the animal under a gauze patch. The patch was placed on the dorso-lateral areas of the trunk of each animal and was held in place with non-irritating tape for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The exposed skin area was carefully washed with water without altering the existing response, or the integrity of the epidermis.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
(indicate if minutes, hours or days): 1, 24, 48, 72 h

SCORING SYSTEM: The degree of erythema/eschar formation and oedema formation was recorded as specified by DRAIZE

Irritation parameter:

edema score

Basis:

animal: #1 - #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

erythema score

Basis:

animal: #1 - #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Other effects:

- Other adverse local effects: not observed
- Other adverse systemic effects: not observed

Irritant Effects on the Skin (Exposure: 4 hours)

Animal 1, Body Weight 2.8 kg Observation (after patch removal)	1 h	24 h	48 h	72 h	day 7	day 14
Erythema (redness) and eschar formation	0	0	0	0
Oedema formation	0	0	0	0
Animal 2, Body Weight 3.1 kg
Observation (after patch removal)	1 h	24 h	48 h	72 h	day 7	day 14
Erythema (redness) and eschar formation	0	0	0	0
Oedema formation	0	0	0	0
Animal 3, Body Weight 2.7 kg
Observation (after patch removal)	1 h	24 h	48 h	72 h	day 7	day 14
Erythema (redness) and eschar formation	0	0	0	0
Oedema formation	0	0	0	0

Summary of Irritant Effects (Score)

Animal		24h	48 h	72 h	Mean scores	Response	Reversible (days)
1	Erythema (redness) and eschar formation	0	0	0	0.0	-	Na
	Oedema formation	0	0	0	0.0	-	Na
2	Erythema (redness) and eschar formation	0	0	0	0.0	-	Na
	Oedema formation	0	0	0	0.0	-	Na
3	Erythema (redness) and eschar formation	0	0	0	0.0	-	Na
	Oedema formation	0	0	0	0.0	-	na
no positive response : mean scores < 2 = -
positive response :mean scores ≥ 2 = +
na: not applicable

Interpretation of results:

GHS criteria not met

Conclusions:

The irritant / corrosive potential of 5-Chlorthiophen-2-carbonsäure was studied on the skin of rabbits. The method used complied with the OECD - Guideline for Testing of Chemicals No. 404 -" Acute Dermal Irritation/Corrosion"; adopted: 24th April 2002.
According to classification criteria 5-Chlorthiophen-2-carbonsäure is not an irritant to the skin. There were no systemic intolerance reactions.

Executive summary:

In a primary dermal irritation study according to OECD guideline 404 (2002), young adult New Zealand White rabbits (3/treatment) were dermally exposed to 0.5 g/patch of Clorthiphen-2-carbonsäure (100 % a.i.) for 4 hours to 5 cm² of shaved dorsal skin. Animals then were observed for 72 h according to the adopted guideline due to the absence of any irritant effect. Irritation was scored as required by the test guideline.

Under the present test conditions all three animals showed no erythema or oedema after patch removal.

All three animals also showed no oedema and erythema at the examination time-points 24 and 48 hours after patch removal.

Based on these results the test item does not need to be classified according to Regulation (EU) No. 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS) with respect to corrosivity.

Reference 2

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-10-10 to 2005-12-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

the study was conducted in accordance to the OECD 431 guideline draft

GLP compliance:

yes

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: not reported

Justification for test system used:

Corrosion of the skin is defined as an irreversible necrotic alteration of the tissue induced by a chemical. While the testing for corrosives usually involves the rabbit skin as a predictive in vivo test method, substances may be classified according to their corrosive potential by the determination of their cytotoxic effects on an in vitro reconstructed human epidermis. The test principle is based on the MTT assay reflecting the cell viability after short term exposure of the epidermal equivalent to topically applied 5-Chlorthiophen-2-carbonsäure.

Vehicle:

physiological saline

Remarks:

0.9% NaCl used to moisted the test item

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: The experiment was carried out on a reconstructed human epidermis EST-1000 (CellSystems, St. Katharinen, Germany).
- Tissue batch number(s): EST-50424-001

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: RT: 3 min; 37 ± 2°C: 60 min
- Temperature of post-treatment incubation (if applicable):

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 3 washing steps, volume not reported
- Observable damage in the tissue due to washing: No

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 2 h
- Spectrophotometer: (EL808, Bio-Tek; 96 well format, 200 µL)
- Wavelength: 570 nm


NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues: not reported
- Procedure used to prepare the killed tissues (if applicable): not reported
- N. of replicates : not reported
- Method of calculation used: not reported

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439: None

Control samples:

yes, concurrent negative control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg


NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL

Duration of treatment / exposure:

3 min and 60 min

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 (60 min exposure)

Value:

28.72

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

not examined

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 (3 min exposure)

Value:

84.93

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

not examined

Compound	Cell viability after 3 min [%]	Cell viability after 60 min [%]	Classification
5-Chlorthiophen-2-carbonsäure	84.93	28.72	Non-Corrosive
Negative control	100.00	100.00	Negative control

Interpretation of results:

GHS criteria not met

Conclusions:

5-Chlorthiophen-2-carbonsäure was characterised by no significant impact on cell viability after 3 min or after the 60 min period. Thus, 5-Chlorthiophen-2-carbonsäure should not be labelled as corrosive to skin.

Executive summary:

In a dermal irritation study performed in accordance with draft OECD Guideline 431 (In Vitro Skin Corrosion, 2005), 5-Chlorthiophen-2-carbonsäure (100% a.i.) was applied to the three-dimensional human epidermis model tissue for an exposure period of 3 and 60 minutes in triplicates. 50 μL of 0.9% NaCl were topically applied to the epidermal surface in order to improve further contact between the solid and the epidermis. Each approximately 25 mg of the test item were applied to the wetted tissues. The test item was spread to match the surface of the tissue.

After 3 minutes exposure at room temperature or 60 minutes exposure in the incubator, the tissues were washed with phosphate buffered saline to remove any residual test material. Cytotoxicity (irritancy) was expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

The negative (0.9% NaCl) control gave responses that were within the acceptance criteria and as such demonstrated the validity of the study.

The relative mean tissue viability obtained after 3 and 60 minutes treatment with 5-Chlorthiophen-2-carbonsäure compared to the negative control tissues was 84.93 and 28.72%, respectively. Since the mean relative tissue viability for the test substance was above 50%, 5-Chlorthiophen-2-carbonsäure is identified to be not corrosive.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-11-03 to 2005-12-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 April 2002

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

HsdIf:NZW

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan France, 03800 Gannat, France)
- Age at study initiation: Young adult animals
- Weight at study initiation: 2.3 kg — 2.5 kg
- Housing: The animals were housed individually in cage units Metall/Noryl by EBECO.
Excrement trays below the cages contained low dust wood granulate bedding (J. Rettenmaier & Söhne, 73494 Rosenberg, Germany).
- Diet (e.g. ad libitum): The animals received the standard diet “SsniffK-Z” 4mm (manufacturer: Ssniff Spezialdiieiten GmbH, 59494 Soest, Germany), approximately 100 g per animal per day
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3
- Humidity (%): 50±25
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other:

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0,1 g

Duration of treatment / exposure:

24 h

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 24h

SCORING SYSTEM: According to DRAIZE

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein: not reported

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 3 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

iris score

Basis:

animal: #1 - #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Irritant / corrosive response data:

There was corneal opacity in one animal with a score of 1.7 which was not reversible during the 21 experimental days, thus, the test item is considered corrosive to the eyes.

Animal		24 h	48 h	72 h	Mean scores	Response	Reversible (days)
1	Corneal opacity	0	2	3	1.7	-	Not revers.
	Iritis	0	0	0	0.0	-	na
	Redness conjunctivae	2	2	2	2.0	-	14
	Chemosis conjunctivae	2	2	1	1.7	-	7
2	Corneal opacity	0	2	2	1.3	-	7
	Iritis	0	0	0	0.0	-	na
	Redness conjunctivae	2	2	2	2.0	-	14
	Chemosis conjunctivae	1	1	0	0.7	-	3
3	Corneal opacity	0	2	2	1.3	-	14
	Iritis	0	0	0	0.0	-	na
	Redness conjunctivae	2	3	2	2.3	-	14
	Chemosis conjunctivae	1	1	1	1.0	-	7
response: corneal opacity : mean scores < 2 = - , ≥ 2 < 3 = +; ≥ 3 = ++ iritis : mean scores < 1 = - , ≥ 1 < 2 = +; = 2 = ++ conjunctival redness : mean scores < 2.5 = - , ≥2.5 = + conjunctival oedema : mean scores < 2 = - ; ≥ 2 = + na : not applicable

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

According to classification criteria 5-Chlorthiophen-2-carbonsäure causes serious damage to eyes without reversibility within 21 days in one animal. There were no systemic intolerance reactions.

Executive summary:

In a primary eye irritation study according to OECD test guideline 405 (2002), 0.1 g 5-Chlorthiophen-2-carbonsäure (100 % a.i.) were instilled into the conjunctival sac of one eye of 3 young adult New Zealand White rabbits for 24 hours. After 24 hours the eye was gently washed with physiological saline. Animals then were observed for 21 days. Irritation was scored by the method of Draize.

After application of 0.1 g of the test sample into the conjunctival sac, there were moderate reactions in the cornea that were not reversible in one animals within 21 days and initially stronger changes in the conjunctivae, which are completely reversible within 14 days. According to Regulation (EU) No. 1272/2008 (CLP) the test item is classified as Category 1 (irriversible effects to the to eye) based on the results obtained under the conditions described.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion:

In a primary dermal irritation study according to OECD guideline 404 (2002), young adult New Zealand White rabbits (3/treatment) were dermally exposed to 0.5 g/patch of Clorthiphen-2-carbonsäure (100 % a.i.) for 4 hours to 5 cm² of shaved dorsal skin. Animals then were observed for 72 h according to the adopted guideline due to the absence of any irritant effect. Irritation was scored as required by the test guideline.

Under the present test conditions all three animals showed no erythema or oedema after patch removal.

All three animals also showed no oedema and erythema at the examination time-points 24 and 48 hours after patch removal.

 

In a dermal irritation study performed in accordance with draft OECD Guideline 431 (In Vitro Skin Corrosion, 2005), 5-Chlorthiophen-2-carbonsäure (100% a.i.) was applied to the three-dimensional human epidermis model tissue for an exposure period of 3 and 60 minutes in triplicates. 50 μL of 0.9% NaCl were topically applied to the epidermal surface in order to improve further contact between the solid and the epidermis. Each approximately 25 mg of the test item were applied to the wetted tissues. The test item was spread to match the surface of the tissue.

After 3 minutes exposure at room temperature or 60 minutes exposure in the incubator, the tissues were washed with phosphate buffered saline to remove any residual test material. Cytotoxicity (irritancy) was expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

The negative (0.9% NaCl) control gave responses that were within the acceptance criteria and as such demonstrated the validity of the study.

The relative mean tissue viability obtained after 3 and 60 minutes treatment with 5-Chlorthiophen-2-carbonsäure compared to the negative control tissues was 84.93 and 28.72%, respectively. Since the mean relative tissue viability for the test substance was above 50%, 5-Chlorthiophen-2-carbonsäure is identified to be not corrosive.

 

Eye irritation/corrosion:

In a primary eye irritation study according to OECD test guideline 405 (2002), 0.1 g 5-Chlorthiophen-2-carbonsäure (100 % a.i.) were instilled into the conjunctival sac of one eye of 3 young adult New Zealand White rabbits for 24 hours. After 24 hours the eye was gently washed with physiological saline. Animals then were observed for 21 days. Irritation was scored by the method of Draize.

After application of 0.1 g of the test sample into the conjunctival sac, there were moderate reactions in the cornea that were not reversible in one animals within 21 days and initially stronger changes in the conjunctivae, which are completely reversible within 14 days.

Justification for classification or non-classification

Based on these results the test item does not need to be classified according to Regulation (EU) No. 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS) with respect to corrosivity to the skin.

According to Regulation (EU) No. 1272/2008 (CLP) the test item is classified as Category 1 (irriversible effects to the to eye) based on the results obtained under the conditions described.