3-amino-N,N-dimethylpropan-1-aminium 2-C10-13-alkyl benzenesulfonate

Endpoint summary

Description of key information

There are modern guideline studies for skin and eye irritation in rabbits. The studies were carried with full GLP compliance and are considered to be Klimisch 1.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study start 20th September 2006 - Report date 16th February 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 (incl. QA statement)

Specific details on test material used for the study:

Identification: Witconate P-1460
Description: Light brown paste
Batch number: 2140-59.01
Activity: 88.8 %
Stability of test item: Stable under storage conditions.
Expiry date: 07-APR-2007
Storage conditions: At room temperature (range of 20 +/-5 °C), light protected

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Source of the rabbits: Harlan Netherlands BV Kreuzelweg 53, NL-5961 NM Horst / The Netherlands, Postbus 6174, NL-5960 AD Horst / The Netherlands
Number of animals per test: 3 (Animals of both sexes were used)
Age at treatment: 13 weeks (male), 10—11 weeks (females)
Identification: By unique cage number and corresponding ear number.
Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
Allocation: Male No. 85, Female Nos. 86, 87

HUSBANDRY
Conditions: Air-conditioned with ranges for room temperature 17-23 °C, relative humidity 30-70 % and approximately 10-15 air changes per hour. Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC. The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.

Accomodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 77/05, Provimi Kliba AG) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance
diet ad Iibitum (batch no. 24/06) provided by Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad Iibitum. Results of analyses for contaminants are archived at RCC Ltd.
Water: Community tap water from Füllinsdorf ad Iibitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.5 g (per animal) of Witconate P-1460 was weighed as delivered by the Sponsor.
The pH of the test item was measured before the study initiation date. A formulation of a 1 % (w/w) solution was prepared. The pH was found to be 5-6.
According to Commission Directive 2004/73/EC, 8.4. and OECD Guidelines 404, a test item needs not to be tested if the pH-value is less than 2 or greater than 11.5, owing to its predictable corrosive properties.

Duration of treatment / exposure:

Four days before treatment, the left flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm x 10 cm). The skin of the animals was examined one day before treatment. and regrown fur of all animals was clipped again.
Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.
On the day of treatment, 0.5 g of Witconate P-1460 was placed on a surgical gauze patch (ca. 4 cm x 4 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.
The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.
As it was suspected that the test item might produce irritancy, a single animal (one female) was treated first. Because moderate signs of irritation were observed following the treatment. the two remaining animals were also treated in a stepwise manner.

Observation period:

Mortality/Viability: Daily from acclimatization of the animals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation.
Clinical signs: Daily from acclimatization of the animals to the termination of test.
Skin reactions: where recorded

Number of animals:

3 (Animals of both sexes were used), Male No. 85, Female Nos. 86, 87

Details on study design:

IRRITATION SCORES
The skin reaction was assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004.
If evident, corrosive or staining properties of the test item were described and recorded.
SKIN OBSERVATION TIME POINTS AND TERMINATION
The skin reaction was assessed at approximately 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after exposure (removal of the dressing, gauze patch and test item).
To allow further examination of the test site, animal No. 85 was re-clipped on completion of the 48 hour examination.

TREATMENT OF RESULTS
Data were summarized in tabular form, showing for each individual animal the irritation scores for erythema and oedema at all measurement intervals. Any lesions were described, including the degree and nature of irritation, corrosion or any other toxic effects observed, and their reversibility.
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. An animal is positive when the mean score is 2 or greater. The test is positive for irritation when at least 2 animals are positive for the same endpoint (erythema/eschar or oedema).

NECROPSY
No necropsy was performed on the animals sacrificed at termination of observation. All rabbits were sacrificed by an intravenous injection of pentobarbitone into the ear vein at a dose of at least 1 mL/kg body weight and discarded.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 10 days

Remarks on result:

probability of moderate irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of mild irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of moderate irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of moderate irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of mild irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of weak irritation

Irritant / corrosive response data:

IRRITATION
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score of the three animals was 2.00, 2.00 and 1.00, respectively and the mean oedema score was 1.00, 1.33 and 0.33, respectively.
A well-defined erythema was observed at the treatment site from the 1-hour reading up to the 24- or 72-hour reading in one and two animals, respectively. A very slight erythema was noted in one animal 24 hours after the treatment and in one animal still at the 7-day reading.
A slight swelling was noted for the three animals one hour after end of the exposure and persisted in one animal still at the 24-hour reading. A very slight swelling was observed 24 to 72 or 48 to 72 hours after the treatment, or, at the 24-hour reading only.
Scaling of the skin at the application site was noted in two of the animals 72 hours to 7 days or 7 to 14 days after the treatment, respectively.

COLORATION

No staining produced by the test item of the treated skin was observed.

CORROSION

Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

VIABILITYIMORTALITYICLINICAL SIGNS

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.


The body weights of all rabbits were considered to be within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), Witconate P-1460 was considered to be "Irritating” to rabbit skin and needed to be labelled with the risk phrase R38 “Irritating to skin”. The classification was based on the EU criteria as two out of the three rabbits had a mean erythema score for 24/48/72 hours of 2. The corresponding EU CLP/GHS criteria for skin irritation requires for the equivalent classification of category 2, a mean value in at least two of the three rabbits over 24/48/72 hours for erythema and or oedema of >2.3-< 4.0. Due to this change in the threshold from 2.0 used in the previous EU criteria for R38 to 2.3 for category 2, Witconate P-1460 does not meet the CLP/GHS criteria to be classified as irritating to skin. It also does not meet the criteria as the effects (erythema and or oedema) did not persist to 14 days with the exception of scaling in one rabbit. For scaling to result in a classification of category 2 for skin irritation it must persist for 14 days in at least 2 of the three rabbits.

Executive summary:

The primary skin irritation potential of Witconate P-1460 was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after removal of the dressing.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately.

The mean erythema/eschar score of the three animals was 2.00, 2.00 and 1.00, respectively and the mean oedema score was 1.00, 1.33 and 0.33, respectively.

The application of Witconate P-1460 to the skin resulted in moderate signs of irritation.

These included erythema, oedema and scaling. These effects were not fully reversible in one of the treated animals during the 14-day observation period. The test item caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed.

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), Witconate P-1460 is considered to be “Irritating" to rabbit skin and needs to be labelled with the risk phrase R38 “Irritating to skin".

The classification was based on the EU criteria as two out of the three rabbits had a mean erythema score for 24/48/72 hours of 2. The corresponding EU CLP/GHS criteria for skin irritation requires for the equivalent classification of category 2, a mean value in at least two of the three rabbits over 24/48/72 hours for erythema and or oedema of >2.3-<4.0. Due to this change in the threshold from 2.0 used in the previous EU criteria for R38 to 2.3 for category 2, Witconate P-1460 does not meet the CLP/GHS criteria to be classified as irritating to skin. It also does not meet the criteria as the effects (erythema and or oedema) did not persist to 14 days with the exception of scaling in one rabbit. For scaling to result in a classification of category 2 for skin irritation it must persist for 14 days in at least 2 of the three rabbits.

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:

Study start 11th October 2006 - Report date 15th February 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 (incl. QA statement)

Specific details on test material used for the study:

Identification: Witconate P-1460
Description: Light brown paste
Batch number: 2140-59.01
Activity: 88.8 %
Stability of test item: Stable under storage conditions.
Expiry date: 07-APR-2007
Storage conditions: At room temperature (range of 20 +/-5 °C), light protected

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test system Young Adult New Zealand White Rabbit, SPF
Rationale Recognized by the international guidelines as the recommended test system.
Source of the rabbits: Harlan Netherlands BV Kreuzelweg 53, NL-5961 NM Horst / The Netherlands, Postbus 6174, NL-5960 AD Horst / The Netherlands
Number of animals per test: 3 (Animals of both sexes were used)
Age at treatment: 12 weeks (male), 13 weeks (females)
Identification: By unique cage number and corresponding ear number.
Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
Allocation: Male No. 88, Female Nos. 89, 90

Conditions: Air-conditioned with ranges for room temperature 17-23 °C, relative humidity 30-70 % and approximately 10-15 air changes per hour. Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC. The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.
Accomodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 77/05, Provimi Kliba AG) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance
diet ad Iibitum (batch no. 24/06) provided by Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad Iibitum. Results of analyses for contaminants are archived at RCC Ltd.
Water: Community tap water from Füllinsdorf ad Iibitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL (per animal) of Witconate P-1460 was measured with a syringe. and applied undiluted as it was delivered by the Sponsor.

Duration of treatment / exposure:

The eye were observed for 14 days after instillation of the test substance.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48, 72 hours , as well as 7, 10 and 14 days after administration.
Eye examinations were made with a Varta Cliptrix diagnostic-lamp (Roth AG, CH-4153 Reinach/Switzerland).

Number of animals or in vitro replicates:

3 rabbits were tested, Male No. 88, Female Nos. 89, 90.

Details on study design:

0.1 mL (per animal) of Witconate P-1460 was measured with a syringe. and applied undiluted as it was delivered by the Sponsor.
The eyes of the animals were examined one day prior to test item administration.
Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of the results were not used in the test.
On the day of treatment, 0.1 mL of Witconate P-1460 was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The test item was applied as a volume instead of weight due to its thick paste-like consistency. The lids were then gently held together for about one second to prevent loss of test item. The right eye remained untreated and served as the reference control. The treated eyes were not rinsed after instillation.

As it was suspected that the test item might produce irritancy, a single animal (one female) was treated first. As neither a corrosive effect nor a severe irritant effect was observed after the 1- and 24-hour examinations, the test was completed using the two remaining animals.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Remarks on result:

probability of mild irritation

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 3 days

Remarks on result:

probability of weak irritation

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of mild irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of mild irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 1 day

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 1 day

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and Chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 2.00, 0.67 and 2.00 for reddening and 1.00, 0.00 and 0.00 for chemosis, respectively.

No abnormal findings were observed in the cornea or iris of any animal at any of the measurement intervals.
A slight and moderate reddening of the conjunctivae was noted in the treated eyes of one and two animals, respectively, one hour after test item instillation. The slight reddening persisted in one animal up to the 48-hour reading whemas a moderate reddening was noted in two animals up to the 72-hour reading and, thereafter, a slight reddening in one animal still at the _7- and 10-day evaluations.

One hour after treatment slight swelling (chemosis) of the conjunctivae was observed in two animals and obvious swelling with partial eversion of lids in one animal. A slight swelling persisted in one animal up to the 72-hour reading.
Moderate reddening of the sclerae was present in the treated eyes of two animals one hour after instillation. The reddening persisted as slight in one animal at the 24-hour heading whereas one animal expressed moderate reddening up to the 48-hour reading and slight redness still up to day 10. Sclera of one animal remained unaffected.

One hour after treatment all three animals were observed with ocular discharge. This was graded slight in one animal, moderate in one animal and marked in one animal, respectively. At the 24-hour reading one animal was still noted with slight discharge.
No abnormal findings were observed in the treated eye of any animal 14 days after treatment, the end of the observation period for all animals.

VIABILITY/MORTALITY/CLINICAL SIGNS

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

COLORATION

No staining produced by the test item of the treated skin was observed.

CORROSION

Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

BODY WEIGHTS

The body weight of the animals was within the range commonly recorded for this strain and age.

Interpretation of results:

GHS criteria not met

Conclusions:

Based upon the referred classification (Commission Directive 200159/EC of August 06, 2001), Witconate P-1460 is considered to be “not irritating” to the rabbit eye. The criteria for classification for eye irritation have since changed with the adoption of CLP (GHS) criteria. For Category 2 eye irritation classification the following criteria apply:
if, when applied to the eye of an animal, a substance produces:
— at least in 2 of 3 tested animals, a positive response of:
— corneal opacity ≥ 1 and/or
— iritis ≥ 1, and/or
— conjunctival redness ≥ 2 and/or
— conjunctival oedema (chemosis) ≥ 2
Calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

Therefore while a score of 2.00 was seen in 2 out of the 3 rabbits the chemosis scores were 1.00 or less and all rabbits recovered fully by day 14. This confirms that Witconate P-1460 should not be classified as a category 2 eye irritant under EU (CLP/GHS)

Executive summary:

The primary eye irritation potential of Witconate P-1460 was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 mL into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after test item instillation.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00  for all three animals. The individual mean scores for the conjunctivae were 2.00, 0.67 and 2.00 for reddening and 1.00, 0.00 and 0.00 for chemosis, respectively.

The instillation of Witconate P-1460 into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible and were no longer evident 14 days after treatment, the end of the observation period for all animals. No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No staining of the treated eyes by the test item was observed and no clinical signs were observed.

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 06, 2001), Witconate P-1460 is considered to be “not irritating” to the rabbit eye.

The criteria for classification for eye irritation have since changed with the adoption of CLP (GHS) criteria. 

Despite this change Witconate P-1460 scores for eye irritation still do not require classification for eye irritation of EU CLP(GHS) category 2.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

The application of Witconate P-1460 to the skin resulted in moderate signs of irritation.

These included erythema, oedema and scaling. These effects were not fully reversible in one of the treated animals during the 14-day observation period. The test item caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed.

Eye Irritation

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 2.00, 0.67 and 2.00 for reddening and 1.00, 0.00 and 0.00 for chemosis, respectively.

Justification for classification or non-classification

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), Witconate P-1460 is considered to be “Irritating" to rabbit skin and needs to be labelled with the risk phrase R38 “Irritating to skin".

The classification was based on the EU criteria as two out of the three rabbits had a mean erythema score for 24/48/72 hours of 2. The corresponding EU CLP/GHS criteria for skin irritation requires for the equivalent classification of category 2, a mean value in at least two of the three rabbits over 24/48/72 hours for erythema and or oedema of >2.3-<4.0. Due to this change in the threshold from 2.0 used in the previous EU criteria for R38 to 2.3 for category 2, Witconate P-1460 does not meet the CLP/GHS criteria to be classified as irritating to skin. It also does not meet the criteria as the effects (erythema and or oedema) did not persist to 14 days with the exception of scaling in one rabbit. For scaling to result in a classification of category 2 for skin irritation it must persist for 14 days in at least 2 of the three rabbits.

The instillation of Witconate P-1460 into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible and were no longer evident 14 days after treatment, the end of the observation period for all animals. No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No staining of the treated eyes by the test item was observed and no clinical signs were observed.

 

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 06, 2001), Witconate P-1460 is considered to be “not irritating” to the rabbit eye.

 

The criteria for classification for eye irritation have since changed with the adoption of CLP (GHS) criteria. Despite this change Witconate P-1460 scores for eye irritation still do not require classification for eye irritation of EU CLP(GHS) category 2.