Xanthan lyase

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 September 2020 - 15 October 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

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)

Version / remarks:

26 June 2020

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of Xanthan Lyase (CAS 113573-69-6, EC name: Xanthan lyase, Enzyme class no 4.2.2.12)

Molecular formula:

n.a.

IUPAC Name:

Active enzyme protein of Xanthan Lyase (CAS 113573-69-6, EC name: Xanthan lyase, Enzyme class no 4.2.2.12)

Constituent 2

Reference substance name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 3

Reference substance name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 4

Reference substance name:

Carbohydrates as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Carbohydrates as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 5

Reference substance name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Test material form:

liquid

Details on test material:

- Lot/batch No.: PPE55581
- Expiration date of the lot/batch: 18 June 2028

Test animals / tissue source

Species:

cattle

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): As supplied

Duration of treatment / exposure:

The exposure period was 10 min.

Duration of post- treatment incubation (in vitro):

120 minutes

Number of animals or in vitro replicates:

Three tissue replicates.

Details on study design:

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: sodium chloride 0.9% w/v

POSITIVE CONTROL USED: ethanol

APPLICATION DOSE AND EXPOSURE TIME: 0.75 mL applied, and corneas were exposed for 10 minutes.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 4 times
- POST-EXPOSURE INCUBATION: 120 minutes

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: light transmission through the cornea
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: As indicated in the TG was used.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

fluorescein leakage

Value:

ca. 0.015

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Value:

ca. 0.6

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

cornea opacity score

Value:

ca. 0.3

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to UN GHS Classification, the test item Xanthan lyase, batch PPE55581 was given No Category classification under the conditions of the test.

Executive summary:

The Bovine Corneal Opacity and Permeability (BCOP) test method was used to evaluate, if Xanthan lyase, batch PPE55581 could induce serious eye damage.

In this test method, damage by the test item is assessed by quantitative measurements of changes in corneal opacity and permeability.

The test was performed according to GLP and in accordance with OECD TG 437.

The undiluted test item was applied for 10 minutes followed by an incubation period of 120 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS).

The In Vitro Irritancy Score of Xanthan lyase, batch PPE55581 was 0.6. 

It was concluded that according to UN GHS Classification, Xanthan lyase, batch PPE55581 was given 'No Category' classification under the conditions of the test.