Tall oil, potassium salt

Endpoint summary

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:

5 September 2005 to 16 September 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.9 to 2.1 kg
- Housing: Individually caged in metal wire cages, 79 x 59 cm bottom area, 38 cm high
- Diet (e.g. ad libitum): maintenance diet for rabbits, rich in crude fibre, ad libitum
- Water (e.g. ad libitum): Tap water, from an automatic watering system, ad libitum
- Acclimation period: 5 days for one animal; 12 days for the two remaining animals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Average of 19.7 °C (continuous control and recording)
- Humidity (%): Average of 71.4 % (continuous control and recording)
- Photoperiod (hrs dark / hrs light): Artificial light from 6 am to 6 pm

IN-LIFE DATES: From: 5 September 2005 To: 16 September 2005

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

First the test material was administered to one animal. As no corrosive effect was observed in the initial test, the test material was administered to two additional animals one week later.

TEST SITE
Hair was clipped on the dorsal areas of the trunks one day before the application of the test material. The test sites were median on the dorsal thoracal region.
The test material was spread on gauze patches in a size of 2.5 cm2 and applied. The gauze was held in place by marginally fixing with non irritating tape and the application sites were covered semi-occlusively by a dressing (self adhesive non woven fabric, hypoallergenic). Access by the animals to the application sites was prevented by a plastic collar.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The tapes with the patches and the collars were removed and the residual test material wiped off using wet cellulose tissue.
- Time after start of exposure: Following the 4 hour exposure period.

OBSERVATIONS
The animals were examined once daily for other than local changes of the skin.
The treated areas were examined for erythema/eschar and oedema as well as for other local alterations (such as hyperplasia, scaling, discolouration, fissures, scabs and alopecia) approximately 1, 24, 48 and 72 hours after patch removal. No further examinations were performed thereafter.
The surrounding of the administration area, i.e. the untreated skin, served as a negative control.

SCORING SYSTEM:
Erythema/Eschar formation
0 No erythema
1 Very slight erythema (barely perceptible)
2 Well-defined erythema
3 Moderate to severe erythema
4 Severe erythema (beet redness) or eschar formation (injuries in depth)

Oedema formation
0 No oedema
1 Very slight oedema (barely perceptible)
2 Slight oedema (edges of area well defined by definite raising)
3 Moderate oedema (raised approximately 1 mm)
4 Severe oedema (raised more than 1 mm and extending beyond area of exposure)

The skin was examined using a cold light source KL 1500 electronic.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

No erythema/eschar or oedema was seen at any test site at any point throughout the observation period.
The control areas were normal at all observations

Other effects:

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.
Residual test material was noted on the exposed skin areas, resp. on the growing hair, until 72 hours post administration.

Table 1 Individual Scores

Time After Exposure (hours)	Erythema/Eschar			Oedema
	Animal Numbers			Animal Numbers
	11	12	13	11	12	13
1	0	0	0	0	0	0
24	0	0	0	0	0	0
48	0	0	0	0	0	0
72	0	0	0	0	0	0
Mean (24, 48 and 72 hours)	0.0	0.0	0.0	0.0	0.0	0.0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the test substance was not irritating to rabbit skin.

Executive summary:

The acute dermal irritation potential of the test substance was investigated according to OECD Guideline 404 and EU Method B.4 under GLP conditions. 0.5 mL of the test substance was applied via a patch to a site about 2.5 cm2 on the intact skin of three New Zealand White rabbits and covered by a semi-occlusive dressing. After an exposure period of 4 hours, any residual test material was removed and the skin examined 1, 24, 48 and 72 hours following patch removal. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. No signs of erythema/eschar or oedema were noted. Under the conditions of this study, the test substance was not irritating to rabbit skin (Wolf, 2005).

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7 December 2010 to 9 December 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes

Species:

other: in vitro reconstructed human epidermis model

Details on test animals or test system and environmental conditions:

The experiment was carried out using reconstructed human epidermis model EST-1000 (CellSystems, St. Katharinen, Germany). The model used for this study has a functional stratum corneum with an underlying layer of living cells.
The tissue equivalents were shipped in 24 well cell culture plates on Agarose supplemented with maintenance medium. Inserts were of 0.6 cm² size.

ENVIRONMENTAL CONDITIONS
The environmental conditions in the incubator were standardised as follows:
- Temperature: 37 ± 2 °C
- CO₂ gas concentration: 5 %. All Incubation steps were performed in a CO₂ atmosphere incubator (Heraeus, Osterode-Germany).
- Humidity: maximum

Type of coverage:

other: not applicable

Preparation of test site:

other: not applicable

Vehicle:

other: The test material was used undiluted, however 30 µL of 0.9 % NaCl was used to moisten the test material and ensure good contact with the cells.

Controls:

other: concurrent in vitro positive and negative controls were used.

Amount / concentration applied:

30 mg

Duration of treatment / exposure:

The cells were exposed to the test material for a period of 20 minutes followed by an incubation period of 42 hours.

Observation period:

Not applicable

Number of animals:

The test was carried out on triplicate tissues.

Details on study design:

POSITIVE AND NEGATIVE CONTROL
-Physiological saline solution (0.9 % NaCl), 30 µL, was used as negative control.
-An SDS (sodium dodecyl sulfate) solution (5 %), 30 µL, was used as positive control.

ADAPTATION TO CELL CULTURE CONDITIONS
Inserts with EST-1000 reconstructed human epidermis (0.6 cm²) were packed under sterile conditions and were shipped refrigerated on supplemented Agarose. Upon arrival, 6 well culture plates were pre-filled with 1 mL of fresh and cool maintenance medium. The reconstructed tissues were placed into the prepared cell culture plates (1 insert/well) and were adapted to the recommended tissue culture conditions (5 % CO₂, 37 °C, max humidity) afterwards for at least 6 hours before use. In case of cultivation for more than 24 hours, a daily medium change is required by aspirating the medium and replacing it with 1 mL of new maintenance medium (37 °C) for each well.

DETERMINATION OF CELL VIABILITY (MTT)
The MTT reduction assay is the most frequently used assay for the determination of cell viability. The assay depends on the intracellular capacity of living cells to chemically reduce the yellow 3-[4,5-Dimethythiazol-2-yl]-2,5-diphenyl-tetrazolium bromide to blue formazan crystals.
The viability of the living cells in the model must be sufficiently high to be able to accurately discriminate between the positive and negative control substances. Cell viability is measured by the amount of MTT reduction, i.e. an OD value following exposure to the negative control or the test material.
A test material is predicted to be a skin irritant if the mean tissue viability is ≤50 %.

After the exposure period of 20 minutes, the inserts were washed carefully in PBS. After a post - exposure incubation of 42 hours, MTT reduction was performed. For viability testing, the inserts were placed in new 24 well plates containing 300 µL of MTT solution (37 °C, 1 mg/mL in MTT-assay medium, delivered by Cell Systems). The tissues were incubated for about 3 hours under cell culture conditions (5 % CO₂, 37 °C, max humidity). The extraction of blue formazan was performed in isopropanol (24 well plates, 2 mL per insert) on a vertical shaker (2 hours). For determination of cell viability, the absorption of the isopropanol-extracts was measured in duplicate at 570 nm in an automatic reader (EL808, Bio-Tek; 96 well format, 200 µL).
Data acquisition and evaluation were done with "Gen5" (software by Bio-Tek).

Irritation / corrosion parameter:

% tissue viability

Value:

84.24

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Table 1 Summary of Results

Sample Number	Material	OD Mean*	Standard Deviation	% Viability
1 to 3	Negative Control	2.32	0.15	100.00
4 to 6	Positive Control	0.02	0.01	1.01
7 to 9	Test Material	1.95	0.13	84.24

*6 values

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the test substance was determined to be a non-irritant.

Executive summary:

The skin irritation potential of the test substance was investigated in an in vitro assay according to OECD Guideline 439 and EU Method B.46 under GLP conditions. The study was conducted using the commercially available reconstructed human epidermis (RHS) model EST-1000. The tissues were exposed to the test material for 20 minutes, followed by a 42 hour incubation period. The cell viability was then assessed in the MTT (3-[4,5-Dimethythiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) conversion assay. Physiological saline was used as negative control, whilst a 5 % sodium dodecyl sulfate solution was used as positive control. The results from these exposures demonstrated the viability and sensitivity of the test model. The viability of the cells exposed to the test material was 84.24 %. Under the conditions of this study, the test substance was determined to be non-irritant to skin (Wingenroth, 2011).

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:

experimental study

Adequacy of study:

key study

Study period:

26 September 2005 to 7 October 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.9 to 2.1 kg
- Housing: Individually caged in metal wire cages, 79 x 59 cm bottom area, 38 cm high
- Diet (e.g. ad libitum): maintenance diet for rabbits, rich in crude fibre, ad libitum
- Water (e.g. ad libitum): Tap water, from an automatic watering system, ad libitum
- Acclimation period: 5 days for one animal; 12 days for the two remaining animals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Average of 19.6 °C (continuous control and recording)
- Humidity (%): Average of 64.9 % (continuous control and recording)
- Photoperiod (hrs dark / hrs light): Artificial light from 6 am to 6 pm

IN-LIFE DATES: From: 26 September 2005 To: 7 October 2005

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and served as control

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

Following administration, the eyes were held closed for about one second to prevent loss of the test material.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

Firstly, the test material was administered to one animal. As no evidence for serious damage to the eye was found during the initial 72 hour observation period (no corrosive effect), the test material was subsequently administered to two further animals.

EYE EXAMINATIONS
Both eyes of the animals were examined within 24 hours before the instillation and approximately 1, 24, 48 and 72 hours post administration.
In addition to the eye examinations, the animals were observed for general changes at all observation times.

SCORING SYSTEM:
CORNEA
Opacity: degree of density (area most dense is taken for reading).
0 No ulceration or opacity
1 Scattered or diffuse areas of opacity (except for slight dulling of normal lustre), details of iris clearly visible
2 Easily discernible translucent area, details of iris slightly obscured
3 Nacreous area, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity

IRIS
0 Normal
1 Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection;
any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive)
2 No reaction to light, haemorrhage, gross destruction (any of all these or all together)

CONJUNCTIVAE
Redness: (refers to the most severe effect of palpebral and bulbar conjunctivae in comparison to the control eye)
0 Blood vessels normal
1 Some blood vessels definitely hyperaemic (injected)
2 Diffuse crimson colour, individual vessels not easily discernible
3 Diffuse beefy red
Chemosis: lids and/or nictating membranes.
0 No swelling
1 Any swelling above normal (includes nictating membranes)
2 Obvious swelling with partial eversion of lids.
3 Swelling with lids about half closed
4 Swelling with lids more than half closed

TOOL USED TO ASSESS SCORE: The entirety of each eye was examined using an otoscope lamp

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

No effects on the cornea or iris were observed in any of the treated eyes. Some conjunctival redness and chemosis were observed at the 1 hour examination only.
The untreated eyes were normal at each observation time.

Other effects:

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1 Individual Scores

Time After Installation (hours)	Cornea			Iris			Conjunctival Redness			Conjunctival Chemosis
	Animal Number			Animal Number			Animal Number			Animal Number
	101	102	103	101	102	103	101	102	103	101	102	103
1	0	0	0	0	0	0	1*	1*	1*	0	1	1
24	0	0	0	0	0	0	0	0	0	0	0	0
48	0	0	0	0	0	0	0	0	0	0	0	0
72	0	0	0	0	0	0	0	0	0	0	0	0
Mean (24, 48 and 72 hours)	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0

*Discharge with moistening of the lids and hairs just adjacent to the lids.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the test substance was not irritating to rabbit eyes.

Executive summary:

The eye irritation potential of the test substance was investigated in vivo according to OECD Guideline 405 and EU Method B.5 under GLP conditions.

0.1 mL of the test material was instilled into the conjunctival sac of the right eye of 3 New Zealand White rabbits. The eyes were examined 1, 24, 48 and 72 hours post administration and the animals observed for general signs of toxicity. The overall mean scores for corneal effects, iris effects, conjunctival redness and conjunctival chemosis were all 0.0. No symptoms of toxicity were observed in the animals. Under the conditions of this study the test substance was not irritating to rabbit eyes (Wolf, 2005).

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

26 January 2011 to 27 January 2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

no guideline available

Principles of method if other than guideline:

The study was carried out using the HCE (Human Corneal Epithelial) model, which is currently undergoing validation conducted by the COLIPA following ECVAM guidelines. The HCE is the only model made from human corneal cells.
Undiluted test material was applied topically to the HCE tissue. After an exposure period of 60 minutes, followed by a post-treatment incubation period of 16 hours, the cell viability was measured by an MTT conversion assay.

GLP compliance:

yes

Species:

other: in vitro human epithelial corneal cell model

Details on test animals or tissues and environmental conditions:

The experiment was carried out on reconstituted human ocular epithelia (SkinEthic Human Corneal Epithelial Model (HCE); SkinEthic, France). The tissue equivalents were shipped in 24 well cell culture plates on semi solid agar's medium. The scope of supply contains Maintenance Medium for incubation (SkinEthic). Inserts were of 0.5 cm² size.

When cultivated at the air-liquid interface in a chemically defined medium, the immortalised human cornea epithelial cells from the cell line HCE reconstruct a corneal epithelial tissue (mucosa), without a stratum corneum, ultra-structurally (tissue morphology and thickness) similar to the corneal mucosa of the human eye.

ENVIRONMENTAL CONDITIONS
The environmental conditions in the incubator were standardised as follows:
- Temperature: 37 ± 2 °C
- CO₂ gas concentration: 5 %. All Incubation steps were performed in a CO₂ atmosphere incubator (Heraeus, Osterode-Germany).
- Humidity: maximum

Vehicle:

other: The test material was used undiluted, however 30 µL of phosphate buffered saline was used to moisten the test material and ensure good contact with the cells.

Controls:

other: concurrent in vitro positive and negative controls were used.

Amount / concentration applied:

30 mg

Duration of treatment / exposure:

The cells were exposed to the test material for a period of 60 minutes, followed by an incubation period of 16 hours.

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

The test was carried out on triplicate tissues.

Details on study design:

POSITIVE AND NEGATIVE CONTROL
-Phosphate buffered saline (PBS), 30 µL, was used as negative control.
-1H-1,2,4-Triazole-3-thiol, 30 mg moistened with 30 µL PBS to ensure good contact with the skin, was used as positive control.

ADAPTATION TO CELL CULTURE CONDITIONS
HCE inserts (0.5 cm²) were packed under sterile conditions and were shipped on semi solid agar's medium. Upon receipt each insert was transferred from the packaging plate to a 6 well culture plate containing 1 mL of fresh maintenance medium per well. The HCE inserts were incubated for at least 2 hours (5 % CO₂, 37 °C, max humidity). Afterwards a media change was performed and the adaptation of the HCE inserts was continued overnight to the recommended tissue culture conditions (5 % CO₂, 37 °C, max humidity). In case of cultivation for more than 24 hours, a daily medium change is required by aspirating the medium and replacing it with 1 mL of new maintenance medium (37 °C) for each well.

DETERMINATION OF CELL VIABILITY (MTT)
The MTT reduction assay is the most frequently used assay for the determination of cell viability. The assay depends on the intracellular capacity of living cells to chemically reduce the yellow 3-[4,5-Dimethythiazol-2-yl]-2,5-diphenyl-tetrazolium bromide to blue formazan crystals.
The viability of the cells in the model must be sufficiently high to be able to accurately discriminate between the positive and negative control substances. Cell viability is measured by the amount of MTT reduction, i.e. an OD value following exposure to the negative control or the test material.
A test material is predicted to be an ocular irritant if the mean relative tissue viability (%) of the exposed cells is ≤50 %.

After the exposure period of 60 minutes, the inserts were washed carefully with PBS. After a post-exposure incubation of 16 hours, the MTT reduction assay was performed. For viability testing the inserts were placed in new 24 well plates containing 300 µL of MTT solution (37 °C, 0.5 mg/mL in Maintenance medium). The tissues were incubated for about 3 hours under cell culture conditions (5 % CO₂, 37 °C, max humidity). The extraction of blue formazan was performed in isopropanol (24 well plates, 1.5 mL per insert) on a vertical shaker (for at least 2 hours). For determination of cell viability per insert, the absorption of the isopropanol-extracts was measured in duplicate at 570 nm in an automatic reader (EL808, Bio-Tek; 96 well format, 200 µL).
Data acquisition and evaluation were done with "Gen5" (software by Bio-Tek).

Irritation parameter:

other: Viability of Cells

Value:

70.54

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritant / corrosive response data:

The viability of the cells exposed to the test material was 70.54 %. The test material was therefore determined to be a non irritant.

The viability of cells exposed to the positive control was 7.54 % (irritant).
The viability of cells exposed to the negative control was 100.00 % (non irritant).

Table 1 Summary of Results

Sample Number	Material	OD Mean*	Standard Deviation	% Viability
1 to 3	Negative Control	1.16	0.21	100.00
4 to 6	Positive Control	0.09	0.02	7.54
7 to 9	Test Material	0.82	0.05	70.54

*6 values

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, the test material was determined to be a non irritant in accordance with EU criteria.

Executive summary:

The potential ocular irritation of the test material was evaluated in an in vitro assay currently undergoing validation under GLP conditions.

The study was conducted by measuring cell viability in the human corneal epithelial cell (HCE) construct, available from SkinEthic Laboratories, following topical exposure to the test material via MTT [3-[4,5-d imethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide] dye conversion.

The model was exposed to the test material for 60 minutes, followed by an incubation period of 16 hours. Cell viability was assessed by an MTT conversion assay.

Phosphate buffered saline was used as negative control, whilst 1H-1,2,4-Triazole-3-thiol was used as positive control. The results from these exposures demonstrated the viability and sensitivity of the test model.

The viability of the cells exposed to the test material was 70.54 %.

Under the conditions of this study, the test material was determined to be a non irritant in accordance with EU criteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation:

Crude tall oil

The acute dermal irritation potential of the test substance was investigated according to OECD Guideline 404 and EU Method B.4 under GLP conditions. 0.5 mL of the test substance was applied via a patch to a site about 2.5 cm2 on the intact skin of three New Zealand White rabbits and covered by a semi-occlusive dressing. After an exposure period of 4 hours, any residual test material was removed and the skin examined 1, 24, 48 and 72 hours following patch removal. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. No signs of erythema/eschar or oedema were noted. Under the conditions of this study, the test substance was not irritating to rabbit skin (Wolf, 2005).

 

Potassium chloride

The skin irritation potential of the test substance was investigated in anin vitroassay according to OECD Guideline 439 and EU Method B.46 under GLP conditions. The study was conducted using the commercially available reconstructed human epidermis (RHS) model EST-1000. The tissues were exposed to the test material for 20 minutes, followed by a 42 hour incubation period. The cell viability was then assessed in the MTT (3-[4,5-Dimethythiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) conversion assay. Physiological saline was used as negative control, whilst a 5 % sodium dodecyl sulfate solution was used as positive control. The results from these exposures demonstrated the viability and sensitivity of the test model. The viability of the cells exposed to the test material was 84.24 %. Under the conditions of this study, the test substance was determined to be non-irritant to skin (Wingenroth, 2011).

 

Eye Irritation

Crude tall oil

The eye irritation potential of the test substance was investigatedin vivoaccording to OECD Guideline 405 and EU Method B.5 under GLP conditions.

0.1 mL of the test material was instilled into the conjunctival sac of the right eye of 3 New Zealand White rabbits. The eyes were examined 1, 24, 48 and 72 hours post administration and the animals observed for general signs of toxicity. The overall mean scores for corneal effects, iris effects, conjunctival redness and conjunctival chemosis were all 0.0. No symptoms of toxicity were observed in the animals. Under the conditions of this study the test substance was not irritating to rabbit eyes (Wolf, 2005).

Justification for classification or non-classification

Based on the in vivo skin and eye irritation studies in New Zealand white rabbits, the test substance does not warrant classification according to EU CLP Regulation (EC) no. 1272/2008.