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Diss Factsheets

Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
no details given
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
publication
Title:
Evaluation of the toxicity of graphene oxide exposure to the eye
Author:
Wu et al.
Year:
2016
Bibliographic source:
Nanotoxicology, 2016; 10(9): 1329-1340

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
not specified
Remarks:
no information on GLP compliance available in this publication

Test material

Constituent 1
Reference substance name:
Reaction product of Graphite, acid-treated and potassium permanganate
IUPAC Name:
Reaction product of Graphite, acid-treated and potassium permanganate
Test material form:
solid: particulate/powder
Details on test material:
Graphite oxide was first prepared using a modified Hummers method from powder graphite (Yan et al., 2013a). In brief, 1.0 g of powdered graphite (Alfa Aesar, 98%) was added to 25 mL of H2SO4 (98%) with continuous stirring for 2h. KMnO4 (3.5 g) was gradually added to the above solution in an ice bath. The resulting mixture was stirred at 37 °C for 2h and diluted with 25 mL of deionized water under vigorous stirring. After adding 10 mL of H2O2 solution (30%) and 80 mL of deionized water, the mixture was filtered and washed with a 10% HCl solution. The paste was dispersed in deionized water to obtain dispersion of graphite oxide. GO suspension (pH 7.00) was obtained via dilution of the as-prepared graphite oxide dispersion with deionized water until the concentration reached ~1 mg/mL, and the above suspension was probe-sonicated for 4h, followed by centrifugation at 12 000 rpm for 10 min to remove any un-exfoliated GO. The resulting supernatant was the GO dispersion.
Specific details on test material used for the study:
GO was prepared from graphite according to a modified Hummer's method, followed by probe sonication of GO dispersion for 4h (Yan et al., 2013a). AFM images showed that the size of GO nanosheets was ~120 nm, and their thickness were no more than 1.2 nm, which suggested that the prepared GO was single-layer nanosheets.
The estimated C/O ratio was ~2.4

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Third Military Medical University (30 Gaotanyan Street, Chongqing) and maintained in pathogen-free conditions
- Age at study initiation: 6 months

Test system

Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
100 µL of GO (100 µg/mL)
Duration of treatment / exposure:
not specified
Observation period (in vivo):
1, 24, 48 and 72 h post GO treatment
Duration of post- treatment incubation (in vitro):
not applicable
Number of animals or in vitro replicates:
3
Details on study design:
The acute eye irritation test was conducted according to OECD Guideline 405 (OECD TG 405, 2012). Three New Zealand White Rabbits (6 months of age) were checked and exhibited no abnormalities in the eyes. One drop of a topical ocular anesthetic (0.4% oxybuprocaine hydrochloride) was applied to each eye 5 min prior to GO application to minimize potential pain and distress. Subsequently, 100 µL of GO (100 pg/mL) were dripped into the conjunctival sac of the right eye after gently pulling the lower lid away from the eyeball. Both lids were then gently held together for approximately 1 s to prevent the loss of GO. The left eye remained untreated and served as the control. The responses, including corneal opacity, conjunctival redness, abnormality of the iris, and chemosis, were observed and graded according to OECD Guideline 405 (OECD TG 405, 2012) in rabbits at 1, 24, 48 and 72 h post GO treatment. The corneal epithelium was stained using fluorescein 72 h after GO treatment.
Twenty microliters and two microliters of 3% fluorescein was dripped into the conjunctival sac of the albino rabbits. The rabbits were examined using slit lamp with cobalt-blue light 2 min later, and photographs were obtained.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
no indication of irritation
Other effects:
no other effects available

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
No indication of any eye irritation potential of graphene oxide in rabbits was found in a study conducted according to OECD 405.
Executive summary:

An in vivo eye irritation test was performed according to OECD Guideline 405 (OECD TG 405 2012) in New Zealand white rabbits by scoring the lesions of conjunctiva, cornea and iris at specific intervals after exposure of the ocular surface to the test item graphene oxide (GO). The eyes of the rabbits were examined at 1, 24, 48, and 72 h after dripping 100 µg/mL GO into the conjunctival sac. No rabbits exhibited corneal opacity, conjunctival redness, abnormality of the iris, or chemosis at any time point after the instillation of GO. A corneal fluorescein staining assay was used to assess GO toxicity to corneal epithelium. There was no impaired corneal epithelium 72 h post-GO treatment. This result indicated that GO did not induce any acute eye irritation or corrosion in rabbits.