2-Pyridinecarboxamide, 4-(4-amino-3-fluorophenoxy)-N-methyl-

Endpoint summary

Description of key information

AFP-Picolinmethylamid revealed neither corrosive nor irritant properties in a human 3D-Skin model as well as no irritant properties in a human 3D-Cornea model.

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

other information

Study period:

Mar 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Principles of method if other than guideline:

The HCE model is currently involved in the eye irritation validation conducted by COLIPA following ECVAM guidelines. Furthermore, it is routinely used by the major Cosmetic and Pharmaceutical companies, and has already been prevalidated in 2004 (van Goethem et. al., Tox in Vitro 20, 2006, 1-17). This model is recognized as the model of choice and scientifically relevant as documented by several publications (Cotovio et. al., Tox in Vitro, 24, 2010, 523-537).

GLP compliance:

yes (incl. QA statement)

Species:

other: human corneal epithelium (HCE)

Strain:

other: not applicable

Vehicle:

other: test item moistened with phosphate buffered saline (PBS, 30 µl)

Controls:

other: negative control: phosphate buffered saline (PBS, 30 µl); positive control: 3-Mercapto-1,2,4-triazole (30 mg, plus 30 µl PBS for moistening)

Amount / concentration applied:

30 mg per insert

Duration of treatment / exposure:

60 minutes

Observation period (in vivo):

post-exposure incubation: 16 hours

Details on study design:

The irritation potential of the test item is assessed by determination of its cytotoxic effect on an reconstituted human ocular epithelia. The test principle is based on the MTT assay reflecting the cell viability after exposure of the cornea equivalent to topically applied test item.
Tests were performed in triplicates. The test item was applied at a 100 % concentration, i.e. 30 mg per insert (plus 30 µl PBS to moisten and ensure good contact to the tissue), for 60 min at room temperature. After the exposure period the inserts were washed carefully with PBS. After a post-exposure incubation of 16 hours in the incubator (37 +/- 2 °C, 5 % CO2, maximum humidity) MTT reduction was performed. Cell viability was measured by the amount of MTT reduction, i.e. an OD value following exposure to the negative or positive control substances or the test item.

Table 1: Summary of results from HCE test with AFP-Picolinmethylamid

Compound	Cell viability [%]	Evaluation
Negative control	100.00	non-irritant
Positive control	9.57	irritant
AFP-Picolinmethylamid	107.60	non-irritant

The test item was detected as non-irritant in this test model.

Interpretation of results:

other: no irritant property

Executive summary:

An in vitro study for assessing ocular irritation was conducted on a human corneal epithelial (HCE) cell model. This model is routinely used by the major Cosmetic and Pharmaceutical companies and has already been prevalidated in 2004 (van Goethem et. al. (2006), Tox in Vitro 20, 1-17). Undiluted AFP-Picolinmethylamid was applied topically to the reconstituted HCE tissue (30 mg per insert plus 30 µl PBS to moisten and ensure good contact to the tissue). After an exposure period of 60 minutes, followed by a 16 hours post-treatment incubation period, the cell viability was 107.60 % as measured by a MTT conversion assay. As the cut-off for a non-irritant to the eye is 50 % (a test substance is predicted to be an ocular irritant if the relative tissue viability (%) exposed to the test substance is < 50 %), AFP-Picolinmethylamid was considered to be non-irritant to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

In two in vitro tests using an artificial 3D-Skin model AFP-Picolinmethylamid revealed neither corrosive (OECD TG 431; Leidenfrost, 2014a) nor irritant properties (OECD TG 439; Leidenfrost, 2014b).

Eye irritation 

AFP-Picolinmethylamid was identified as non-irritant to human corneal epithelial cells (cell viability = 107.60 %) in the in vitro HCE test

(Wingenroth, 2014).

Justification for selection of skin irritation / corrosion endpoint:
No study was selected, since both in vitro studies with AFP-Picolinmethylamid revealed no adverse effects in a skin model.

Justification for selection of eye irritation endpoint:
Only one study available

Justification for classification or non-classification

Based on the study results a classification according to Directive 67/548/EEC or Regulation (EC) No. 1272/2008 (CLP) is not required.