L-Glutamic acid, N-[4-(4-oxobutyl)benzoyl]-, 1,5-diethyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 3 to 4 December 2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Some deviation to Good Lab Practices. Study director was not supplied with test article characterisation information. Therefore the effect of the lack of test article characterization cannot be fully assessed.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,5-diethyl (2S)-2-{[4-(4-oxobutyl)phenyl]formamido}pentanedioate

EC Number:

700-603-5

Cas Number:

908145-87-9

Molecular formula:

C20 H27 N O6

IUPAC Name:

1,5-diethyl (2S)-2-{[4-(4-oxobutyl)phenyl]formamido}pentanedioate

Details on test material:

Analytical purity - assume 100% potency
Lot Number Z84-H70580-095

Test animals / tissue source

Species:

other: Bovine

Strain:

not specified

Details on test animals or tissues and environmental conditions:

This is an in vitro study. Bovine eyes were received from Spear Products on 4 December 2009.

Test system

Vehicle:

water

Amount / concentration applied:

Corneas were dosed with 0.75ml of a mixture of Intermediate 2945989 and MEM (Minimum Essential Media) solution. Constituents of the MEM solution are - MEM powder; sodium bicarbonate; L-Glutamine; Fetal Bovine serum & distilled water.

Duration of treatment / exposure:

Four hours + 90 minutes

Observation period (in vivo):

two hours

Number of animals or in vitro replicates:

n/a this is an in vitro study

Details on study design:

Two corneas were dosed only with the MEM solution to act as controls and placed in an incubator at 32oC. The Intermediate 2945989 MEM solution mixture was applied to five test corneas, which were also placed in an incubator. After four hours both the controls and the test articles were washed with pure MEM solution so that opacity measurements could be taken.

Sodium fluorescein dye in a phosphate buffered saline was then applied to both the controls and test corneas, which were placed in the incubator at 32oC for a further 90 minutes. Optical density of the fluid removed from the cornea holders of both the controls and test articles was measured using a spectrophotometric analysis.

The corrected mean opacity score was calculated using the control and treated cornea opacity values as determined from the OP-KIT. The corrected Mean Optical Density Score was calculated using the control and treated Opacity Density Values from the fluorescein permeability analysis. The in vitro score was calculated as follows:
In Vitro Score = Corrected Mean Opacity Score + 15 (Corrected Mean Optical Density Score)

Results and discussion

In vivo

Results

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: In vitro score

Score:

ca. 1.97

Reversibility:

not specified

Remarks on result:

other: General classification scheme of Gautheron et al. (1992)

Irritant / corrosive response data:

Corrected mean opacity score was 1.5.
Corrected mean optical density (permeability) score was 0.031.
Therefore the in vitro score was calculated as 1.97

Other effects:

No other additional effects were observed during the course of this study.

Any other information on results incl. tables

Results after treatment with the test substance:

Cornea #	O.D. Scores	Control Score	Corrected O.D. Score
1	0.025	0.018	0.007
2	0.125	0.018	0.107
3	0.028	0.018	0.010
4	0.034	0.018	0.016
5	0.031	0.018	0.013
Corrected Mean Optical Density Score=				0.031
Corrected Mean Opacity Score =				1.5
Calculated In Vitro Score 1.5 + 15 (0.031) = 1.97

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: General classification scheme of Gautheron et al. (1992)

Conclusions:

Under the experimental conditions of this study, Intermediate 2945989, is considered non-irritating to the eyes.

Executive summary:

Five corneas were dosed with 0.75 ml of a 20% suspension of Compound 2945989, Lot# Z84-H70580-095. Opacity measurements and sodium fluorescein permeability were determined.

The corrected mean opacity score was 1.5. The corrected mean optical density (permeability) score was 0.031.

The in vitro score was calculated as 1.97, therefore Compound 2945989, Lot# Z84-H70580-095 is considered to be non-irritating.