Oxidation products of D-Glucose with nitric acid, sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Oxidation products of D-Glucose with nitric acid, sodium salts

Molecular formula:

Reaction products consisting mainly of sodium salts of D-glucarate, mono, di and tri carboxylic acids and keto gluconates. See remarks

IUPAC Name:

Oxidation products of D-Glucose with nitric acid, sodium salts

Test material form:

liquid

Specific details on test material used for the study:

D-Glucose, reaction products with nitric acid and sodium nitrite (1:1), sodium salts

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Isolated bovine cornea obtained from freshly slaughtered animals, age 12-48 months. Whole eyes were transported to the test facility in HBSS on ice. The eyes were carefully examined for pre-existing defects and used within 12 hours of collection.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

0.75 mL, undiluted

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

2 hours

Number of animals or in vitro replicates:

5 (negative control), 3 (test material) or 4 (positive control) replicates

Details on study design:

SELECTION AND PREPARATION OF CORNEAS

Isolated bovine cornea obtained from freshly slaughtered animals, age 12-48 months. Whole eyes were transported to the test facility in HBSS on ice.

QUALITY CHECK OF THE ISOLATED CORNEAS

The eyes were carefully examined for pre-existing defects and used within 12 hours of collection.

NUMBER OF REPLICATES

5 (negative control), 3 (test material) or 4 (positve control) replicates

NEGATIVE CONTROL USED

Water for irrigation

POSITIVE CONTROL USED

Ethanol

APPLICATION DOSE AND EXPOSURE TIME

0.75 mL; 10 minutes

TREATMENT METHOD

Closed chamber

REMOVAL OF TEST SUBSTANCE

- Number of washing steps after exposure period: 3

- POST-EXPOSURE INCUBATION

2 hours

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: illuminance
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)
- Other: visual observation

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: decision criteria as indicated in the TG was used.

Results and discussion

In vitro

Results

Irritation parameter:

cornea opacity score

Run / experiment:

Mean IVIS from three replicates

Value:

2.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Mean IVIS 38.45

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: none

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

In vivo

Other effects:

None observed

Any other information on results incl. tables

Test material	Mean IVIS 2.3
Positive control	Mean IVIS 38.45

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on a mean IVIS score of 2.3, the test material [D-Glucose, reaction products with nitric acid and sodium nitrite (1:1), sodium salts] does not require classification for serious eye damage or eye irritation according to the CLP Regulation.

Executive summary:

The potential of the test material [D-Glucose, reaction products with nitric acid and sodium nitrite (1:1), sodium salts] to cause serious eye damage was assessed in a BCOP assay (closed chamber method) performed to OECD 437. 0.75 mL of the test material was applied to the cornea (three replicates) for 10 minutes, followed by a 2-hour post-exposure incubation period. Corneal opacity was measured; corneal permeability was assessed using sodium fluorescein solution and measurement of absorption at 490 nm and IVIS scores calculated. Isolated corneas were also exposed in parallel to negative control (water) and positive control (ethanol) substances. A mean IVIS score of 2.3 was calculated for the test material, indicating that it does not require classification for serious eye damage or eye irritation according to the CLP Regulation. Responses to the negative and positive controls confirmed the integrity and sensitivity of the assay.