2-phosphonobutane-1,2,4-tricarboxylic acid

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Materials and methods

Principles of method if other than guideline:

In vitro study for assessing ocular irritation of compounds using a human epithelial corneal cell model. The model used is standardized and commercially available (SkinEthic™ Human Corneal Epithelial Model (HCE); SkinEthic, France).

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-phosphonobutane-1,2,4-tricarboxylic acid

EC Number:

253-733-5

EC Name:

2-phosphonobutane-1,2,4-tricarboxylic acid

Cas Number:

37971-36-1

Molecular formula:

C7H11O9P

IUPAC Name:

2-phosphonobutane-1,2,4-tricarboxylic acid

Test material form:

solid

Details on test material:

- Purity: ca. 86 % (study report LNX31929, dated 2011-09-05, Spectral Services , D-50996 Köln, Germany)
- The substance is anhydrous.

Test animals / tissue source

Species:

other: not applicable - human corneal epithelial model (HCE)

Strain:

other: not applicable - human corneal epithelial model (HCE)

Test system

Vehicle:

other: not applicable - human corneal epithelial model (HCE)

Controls:

other: not applicable - human corneal epithelial model (HCE)

Duration of treatment / exposure:

After an exposure period of 60 minutes, followed by a 16 hours post-treatment incubation period, the cell viability was measured

Observation period (in vivo):

not applicable - human corneal epithelial model (HCE)

Number of animals or in vitro replicates:

not applicable - human corneal epithelial model (HCE)

Results and discussion

In vitro

Results

Irritation parameter:

other: cell viability

Run / experiment:

60 minutes followed by a 16 h post-treatment period

Value:

1.98

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: irritant - scores in [%] viability

Any other information on results incl. tables

A test substance is predicted to be an ocular irritant if the mean relative tissue viability (%) exposed to the test substance is<50%.

As can be seen from the information in the Table below the test substance

Bayhibit AM wasserfrei was detected as irritant to the cornea epithelium in this test model.

Compound              Cell viability [%] Evaluation

Bayhibit AM wasserfrei      1.98        irritant

Positive control               28.91      irritant

Negative control              100.00      non-irritant

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Executive summary:

This in vitro study for assessing ocular irritation of compounds using a human epithelial corneal cell model was performed in 2011. The model used is standardized and commercially available (SkinEthicTM Human Corneal Epithelial Model (HCE); SkinEthic, France).

Undiluted Bayhibit AM wasserfrei was applied topically to the HCE tissue, i.e. 30 mg per insert. (plus 30 µl PBS to moisten and ensure good contact with the skin; three replicates)

After an exposure period of 60 minutes, followed by a 16 hours post-treatment incubation period, the cell viability was 2% (rounded) as measured by a MTT conversion assay.

The results of the concurrent negative (NC, PBS) and positive control

(PC, 1H-1,2,4-Triazole-3-thiol) demonstrated the viability (NC) and sensitivity (PC) of the test model.

The results show that Bayhibit AM wasserfrei is predicted as ocular irritant under the conditions of this test method.