By-product from Guanidinoacetic acid manufacturing

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28.01.2021 - 2021-03-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

By-product from Guanidinoacetic acid manufacturing

EC Number:

951-773-4

Molecular formula:

- Hydantoic Acid Sodium salt: C3H5N2O3Na - Cyanoguanidine: C2H4N4 - Sodium acetate: C3H3O2Na - Glycine: C2H5NO2 - N,N'-Guanidinodiacetic acid: C5H9N3O4 - N,N-Guanidinodiacetic acid: C5H9N3O4 - Guanidinoacetic acid:C3H7N3O2 - Urea: CH4N2O - Water: H2O - Biguanidinoacetic acid: C4H10N6O2 - Sodium formate: CHO2Na - Glycylglycine: C4H8N2O3

IUPAC Name:

By-product from Guanidinoacetic acid manufacturing

Test material form:

solid

Specific details on test material used for the study:

201009MA2 solid

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered at the abattoir A. Moksel AG, Buchloe, Germany.
On the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories. Immediately after arrival of the eyes, cornea preparation was initiated.

Test system

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

The test item was suspended with physiological saline 0.9% NaCl to give a 20% concentration

Duration of treatment / exposure:

4 h ± 5 min

Number of animals or in vitro replicates:

3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive control treated with imidazole 20% in physiological saline 0.9% NaCl

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

ca. 4.1

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

With an IVIS > 3

Conclusions:

No stand-alone prediction can be made regarding the classification of the test substance By-product from Guanidinoacetic acid manufacturing according to the evaluation criteria. Further testing in another suitable method is required.

Executive summary:

The eye irritancy potential of By-product from Guanidinoacetic acid manufacturing was investigated in the bovine corneal opacity and permeability assay.

Preparation of the test item: The test item was suspended with physiological saline 0.9% NaCl (see 10.3) to give a 20% concentration.
Visual observation after treatment: None of the corneas treated with By-product from Guanidinoacetic acid manufacturing showed any opacity of the tissue.

Mean in vitro irritation score: 4.10
Classification:

☐ UN GHS No Category
☒ no stand-alone prediction can be made
☐ UN GHS Category 1

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.
The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.