Acid D,L-aspart

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented study according to international accepted guidelines and GLP compliant.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not applicable

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Acid D,L-aspart

EC Number:

210-513-3

EC Name:

Acid D,L-aspart

Cas Number:

617-45-8

Molecular formula:

C4H7NO4

IUPAC Name:

aspartic acid

Test material form:

solid: crystalline

Details on test material:

Test item: D,L-aspartic acid
Alternative name: D,L-aszparaginsav cfn. 100% Hat.
Batch No.: S27170N
Physical state: Solid
Loss on drying: 13.5%
Active ingredient in dry substance: 98.3%
Chloride ion: 200 µg / g
Manufacturing date: 20 July 2012
Expiry date: 20 January 2014
Storage: 15-30°C

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species and strain: New Zealand white rabbit
Source: S & K-LAP Kft. 2173 Kartal, Császár út 135, HUNGARY
Hygienic level during the study: good conventional
Justification of strain: The New Zealand white rabbit is one of the standard animals in acute eye irritation study.
Number of animal: 3 males
Age of animals: adult rabbits
Body weight range at the
beginning of the study: 3170 - 3430 g
Body weight range at the at the end of the study: 3331 - 3720 g
Date of receipt: November 15, 2012
Acclimatisation time: 27 days
Animal health: Only animal in acceptable health condition was used for the test.
Room: 1 (F building)
Housing: Animal was housed individually in metal cage.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20 ± 3 °C
Relative humidity: 30-70 %
The environmental parameters were recorded daily during the study. Before housing the animals the microbiological status of the room was checked. The animals received Rabbit fattening mixed diet produced by YAQ-TÁP Kft., Nyíregyháza (Tokaji út 22), Hungary, ad libitum. Copies of the Certificates of food are maintained in Toxi-Coop Zrt.’s archive. Animals received tap water from watering bottles ad libitum. The drinking water is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive. The individual identification was performed by numbered ear tag. The cages were marked with individual identity cards with information about study number, dose group, sex, cage number and individual animal number.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The contralateral eye served as control.

Amount / concentration applied:

0.1 g of the test item D,L-aspartic acid was used for the study in undiluted state, in a single dose. The test item was powered by mortar before treatment. The absorption of the test item was not determined.

Duration of treatment / exposure:

The eyes of the test animals were not washed out after the application of test item.

Observation period (in vivo):

The eyes were examined at 1, 24, 48 and 72 hours after the application. The duration of the observation period was sufficient for the statement of reversibility or irreversibility of changes.

Number of animals or in vitro replicates:

3

Details on study design:

Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition, any with ocular lesions were rejected. In the first step an initial test was performed using one animal. The test item was poured into the conjunctival sac of the left eye. The eyelids were held closed gently for several seconds to prevent the loss of the test item. The contralateral eye served as control. Immediately after the administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix V. After consideration of the ocular responses produced in the first animal, two additional animals were treated. Before the administration the treated eyes of animals were not anaesthetised, because the score of initial pain reaction was 2 in the first animal.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24h;48h;72h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24h;48h;72h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24h;48h;72h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

other: discharge score

Basis:

mean

Time point:

other: 24h;48h;72h

Score:

0

Max. score:

0

Results 5

Irritation parameter:

other: redness

Basis:

mean

Time point:

other: 24h;48h;72h

Score:

0

Max. score:

0

Irritant / corrosive response data:

The eyes were examined at 1, 24, 48 and 72 hours after the application. One hour after the treatment some hyperemic blood vessels (score 1) occurred in two animals (animal No.: 01316, 01346). The discharge with moistening of the lids and hairs just adjacent to lids (score 2) was observed in all animals. 24 hours after the treatment all animals were symptom-free. 48 hours after the treatment no primary irritation symptoms occurred. 72 hours after the treatment the study was terminated, since no primary irritation symptoms occurred. During the study the control eyes of animals were symptom-free. General state and the behaviour of animals were normal throughout the study period. There were no notable body weight changes during the contact and observation period.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, test item D,L-aspartic acid applied to the rabbits' eye mucosa, caused slight to moderate conjunctival irritant effects, fully reversible within 24 hours. According to the EC criteria for classification and labelling requirements for dangerous substances and preparations, the test item does not have to be classified and has no obligatory labelling requirement for eye irritation. According to Regulation (EC) No. 1272/2008, the test item has not been classified into any category.