Glycine, N-acetyl-N-[3-(trifluoromethyl)phenyl]valyl-, ethyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 13 August 2002 to 09 December 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24th February 1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

31st July 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Glycine, N-acetyl-N-[3-(trifluoromethyl)phenyl]valyl-, ethyl ester

EC Number:

609-496-9

Cas Number:

379685-94-6

Molecular formula:

C18H23F3N2O4

IUPAC Name:

Glycine, N-acetyl-N-[3-(trifluoromethyl)phenyl]valyl-, ethyl ester

Test material form:

solid: particulate/powder

Details on test material:

- Other: ochre fine powder

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: room temperaturepH OF TEST ITEM: approximately 5 at the concentration of 10% in purified water, measured at thetesting laboratory.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS- Source: CEGAV, Saint Mars d'Egrenne, France.- Age at study initiation: 2 to 4 months old- Weight at study initiation: 3.1 ± 0.1 kg- Housing: animals were housed individually in polystyrene cages (48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.- Diet: ad libitum to 110 pelleted diet (UAR, Villemoisson, Epinay-sur-Orge, France). Diet was analysed regularly by the supplier for composition and contaminant levels (dietformula was available in the study report).- Water: ad libitum to drinking water filtered by a FG Millipore membrane (0.22 micron). Bacteriological and chemical analyses of water were performed regularly by extemal laboratories (include detection of possible contaminants (pesticides, heavy metals andnitrosamines)).- Acclimation period: at least five days.ENVIRONMENTAL CONDITIONS- Temperature (°C): 18±3°C- Humidity (%): 30 to 70%- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air.- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the untreated right eye served as control

Amount / concentration applied:

0.1 g of the test item in its original form

Observation period (in vivo):

1 hour, 24, 48 and 72 hours after administration of the test item. As no persistent ocular reaction was obserevd, the study was ended on day 4.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

PRIOR TO TREATMENT: both eyes of each animal were examined 21 hours before tratment in order to reject any animal showing signs of ocular lesions, ocular defects of pre-existing corneal injury.ADMINISTRATION OF TEST SUBSTANCE: As no irritant effects effects were anticipated, the study was directly performed on three animals. the test substance was introduced in its original form into the conjunctival sac of the left eye after gently pulling the lower lid away from the eyeball. After the application, the lower and upper eyelids were held together for approximately one second to avoid any loss of test item.REMOVAL OF TEST SUBSTANCE- Washing (if done): no washingEVALUATION OF OCULAR REACTIONS:Conjunctival reactions, iritis and comeal opacification were evaluated daily for each animal. TOOL USED TO ASSESS SCORE: For the evaluation of comeal opacification (presence or absence, affected area), the eyes were examined under a UV lamp after instillation of one or two drops of 0.5% sodium fluorescein solution (a clear fluorescence is visible in the areas of opacification). This evaluation was performed on day 2 and repeated thereafter whenever necessary.SCORING SYSTEM:Ocular reactions were scored according to the following numerical scale: Coniunctival lesions and dischargeChemosis (lids and/or nictitating membranes) . no swelling...............................................................................................................................0 . any swelling above normal (includes nictitating membranes)................................................. 1 . obvious swelling with partial eversion of lids..........................................................................2 . swelling with lids about half-closed.........................................................................................3 . swelling with lids more than half-closed..................................................................................4 Redness (refers to palpebral and bulbar conjunctivae, comea and iris) . bloodvessels normal................................................................................................................0 . a number ofblood vessels definitely hyperemic (injected)...................................................... 1 . diffuse, crimson colour, individual vessels not easily discemible ...........................................2 . diffuse, beefy red......................................................................................................................3 Discharge . absence of discharge.................................................................................................................0 . slight discharge (does not include small amounts normally found in inner canthus)........................................................................................................................... 1 . discharge with moistening of lids and hairs adjacent to lids....................................................2 . discharge with moistening of lids and hairs on wide area around the eye ...............................3 Iris lésions . normal ......................................................................................................................................0 . markediy deepened rugae, congestion, swelling, moderate circum-comeal hyperemia, or injection, any of thèse or combination ofany thereof, iris still reacting to light (sluggish reaction is positive) ........................................................................ 1 . no reaction to light, haemorrhage, gross destruction (any or ail of thèse) ...............................2

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 0 was obtained at the 1h observation time point

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 0 was obtained at the 1h observation time point

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 0 was obtained at the 1h observation time point

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 0 was obtained at the 1h observation time point

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 0 was obtained at the 1h observation time point

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 0 was obtained at the 1h observation time point

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48h

Remarks on result:

other: a score of 2 was obtained at the 1h observation time point

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48h

Remarks on result:

other: a score of 2 was obtained at the 1h observation time point

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48h

Remarks on result:

other: a score of 2 was obtained at the 1h observation time point

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48h

Remarks on result:

other: a score of 1 was obtained at the 1h observation time point

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: a score of 2 was obtained at the 1h observation time point

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48h

Remarks on result:

other: a score of 1 was obtained at the 1h observation time point

Irritant / corrosive response data:

Very slight or slight conjunctival reactions, including a very slight or slight chemosis (grade 1 or 2), a very slight or slight redness of the conjunctiva (grade 1 or 2) and a clear discharge (grade 1 or 2), were observed in all animals, at 1h and 24 hours only. No other ocular reactions were observed during the study. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.00, 0.00 and 0.00 for comeal opacity, 0.00, 0.00 and 0.00 for iris lesions, 0.33, 0.33 and 0.33 for redness of the conjunctiva and 0.33, 0.00 and 0.33 for chemosis.

Other effects:

None.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the test material is not classified as irritating to the eye according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) criteria.

Executive summary:

This GLP-compliant study was performed to assess the potential of the test material to induce eye irritation in vivo, according to OECD Guideline 405 (dated February 24th, 1987) and EU method B.5 of the E.E.C. directive n° 92/69 (dated July 31st, 1992).

Material and methods

0.1 g of the test material were introduced as supplied, into one eye of each of the 3 male rabbits, the other untreated seye erving as control. Ocular examinations were performed on both right and left eyes 1 hour

and then 24, 48 and 72 hours after instillation.

Results

Ocular reactions (redness of the conjunctivae, chemosis and discharge) were observed during the study but were all fully reversed 48 hours after the administration of the test material. The individual mean scores for each animal (24, 48 and 72 hours) were 0.00, 0.00 and 0.00 for comeal opacity, 0.00, 0.00 and 0.00 for iris lesions, 0.33, 0.33 and 0.33 for redness of the conjunctiva and 0.33, 0.00 and 0.33 for chemosis.

Conclusion

Under the experimental conditions of this study, the test material is not classified as irritating to the eye according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) criteria.