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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
review article or handbook
Title:
RTECS Number: BY1575000
Year:
2012
Bibliographic source:
RTECS (Registry of Toxic Effects of Chemical Substances):- Journal of Toxicologic Pathology. (Nihon Dokusei Byori Gakkai, editor, 3-25-8 Nishi- shinbashi, Minato-ku, Tokyo 105, Japan) V.1- 1988 Volume(issue)/page/year: 29,187,2001

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other:
Principles of method if other than guideline:
Low-Volume Eye Test: Ten μL of parafluoroaniline (PF) was directly applied to the cornea of the right eye of each rabbit. Eyes and eyelids were macroscopically scored for signs of irritation beginning 3 hours after dosing and periodically until recovery or 35 days. Tissues were obtained for light microscopic examination after 3 hours and on days 1, 3, and 35.
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: 4-fluoroaniline
- Molecular formula: C6H6FN
- Molecular weight: 111.12
- Substance type: Organic
- Physical state: Liquid

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
10 µL
Duration of treatment / exposure:
the animals were treated once and eyes were examined macroscopically and scored for irritation at 3 hours and 1, 3, 7, and 35 days.
Observation period (in vivo):
35 days
Number of animals or in vitro replicates:
6
Details on study design:
Eyes were macroscopically examined with the aid of a penlight before dosing to insure absence of ocular abnormalities. Ten µL of an irritant was taken up in a 100 µL Teflon tipped Hamilton syringe (Hamilton Co, Reno, NV) and excess test substance was removed from the exterior of the syringe. The eyelids were then gently held open and 10 µL was placed directly above the center of the cornea of the right eye. Eyelids were released immediately after instillation without forced blinking or manipulation.
Eyes and eyelids were macroscopically scored for signs of irritation beginning 3 hours after dosing and periodically until recovery or 35 days. Tissues were obtained for light microscopic examination after 3 hours and on days 1, 3, and 35.
In vivo Confocal Microscopy assessment of the corneas was performed after each macroscopic examination and separately at 14 days. After evaluation at 35 days, the animals were euthanatized.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Basis:
mean
Remarks:
by Light microscopy
Time point:
other: 3 hr
Score:
53.5
Max. score:
110
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Moderate to severe irritation; Days to recovery >35 days
Irritation parameter:
overall irritation score
Basis:
mean
Remarks:
In vivo CM and Live–Dead Assay
Time point:
other: 3 hr
Score:
53.4
Max. score:
110
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Moderate to severe irritation; Days to recovery >35 days
Irritation parameter:
maximum mean total score (MMTS)
Remarks:
Conventional MAS: Maximum Average Score
Basis:
mean
Time point:
other: observation periods beginning at day 1.
Score:
80
Max. score:
110
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Moderate to severe irritation; Days to recovery >35 days

Applicant's summary and conclusion

Interpretation of results:
moderately irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
10 µL of 4-fluoroaniline instilled into rabbit eye caused moderate irritation as evaluated by a Low-Volume Eye Test
Executive summary:

Low-Volume Eye Test was conducted to determine theon the extent of initial injury caused by 4 -fluoroaniline.Ten μl of parafluoroaniline (PF) was directly applied to the cornea of the right eye of each rabbit. Eyes and eyelids were macroscopically scored for signs of irritation beginning 3 hours after dosing and periodically until recovery or 35 days.Tissues were obtained for light microscopic examination after 3 hours and on days 1, 3, and 35. Initial corneal injury was characterized quantitatively at 3 hours and 1 day using in vivo confocal microscopy (CM) and by postmortem quantitation of dead corneal epithelial cells and keratocytes using a Live—Dead Assay (L/D, Molecular Probes) and scanning laser CM. Corneal changes over time were characterized quantitatively using in vivo CM performed at 3 hours and 1, 3, 7, 14, and 35 days.

Changes with parafluoroaniline were consistent with moderate to severe irritation.Specifically,corneal injury by parafluoroaniline tended to involve the epithelium and anterior stroma, with the mean normalized depth of injury (NDI) being 10.4% to 23.8%. The results can be summarised as:

  • Overall irritation score by light microscopy ranged between 53.5 - 110 after 3 hours.
  • Overall irritation score calculated as a mean of in-vivo Cm and live-dead assay ranged between 53.4 - 110 after 3 hours.
  • Maximum Mean Total Score (MMTS) ranged between 80 - 110 after 24 hours. tal Score (MMTS) rangd between 80 - 110 after 24 hours.
  • All the irritation effects were not fully reversible within 35 days.