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

Eye irritation

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

824-801-7

Cas Number:

1093628-27-3

Molecular formula:

not applicable

IUPAC Name:

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

Test material form:

solid

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

Test animals / tissue source

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.

Test system

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.

Results and discussion

In vivo

Results

Results 1

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

Results 2

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

Results 3

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

Results 4

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

Results 5

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

Results 6

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

Results 7

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

Results 8

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

Results 9

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

Results 10

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

Results 11

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

Results 12

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.

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.