Benzoic acid, 4-(methylamino)-3-nitro-

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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

The experimental design of the study consists of a test for Direct Reduction of MTT by the test material, followed by the main test.
For the main test, triplicate SkinEthic tissues were treated with 30 mg oftest material and exposed for 10 minutes and 60 minutes. The tissues were
incubated at 37°C in a humidified atmosphere of 5% C02 in air for the appropriate exposure period.
Duplicate tissues served as the negative control or positive control for each exposure period.
At the end of the 10 minute exposure period each SkinEthic tissue was rinsed using Dulbecco' s Phosphate Buffered Saline (DPBS) and placed into
a 'holding plate' until all the tissues had been rinsed. The rinsed tissues (two per group) were then transferred to an MTT 'loading plate', and
incubated at 37°C for approximately 3 hours in a humidified atmosphere of 5% C02 in air. The remaining test material treated tissue was retained for possible histology. Following MTT loading, each SkinEthic tissue was blotted dry and placed into an MTT 'extraction plate' in order to extract aIl of the reduced MTT from the tissues. The same rinsing, retention, loading and extraction procedures were repeated for the 60 minute tissues once
the 60 minute exposure period was complete.
At the end of the extraction period, the extracted MTT solution was mixed for each SkinEthic tissue and 3 x 200 111 sampies, representing each
tissue, were transferred to the appropriate weIls of a 96 weIl plate. The optical density at 540nm (ODS40) of each weIl was measured.
Data arepresented in the form of % viability (MTT conversion relative to negative controls) for each of the two exposure periods.
The results were used to make a prediction of the eye irritation potential of the test material.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-(methylamino)-3-nitrobenzoic acid

EC Number:

609-904-5

Cas Number:

41263-74-5

Molecular formula:

C8 H8 N2 O4

IUPAC Name:

4-(methylamino)-3-nitrobenzoic acid

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

other: relative mean viability [%]

Value:

92.7

Remarks on result:

other: exposure time: 10 minutes

Results 2

Irritation parameter:

other: relative mean viability [%]

Value:

92

Remarks on result:

other: exposure time: 60 minutes

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Negative result, not likely to be a severe ocular irritant.