Graphite

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-03-10 to 2010-07-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well performed GLP study according to recent OECD guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

2008

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate attached to full study report

Test material

Test material information

1

Reference substance name:

Graphite

EC Number:

231-955-3

EC Name:

Graphite

Cas Number:

7782-42-5

Molecular formula:

C

IUPAC Name:

carbon

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

other: Russian Crl:CHBB

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation: approx. 28 - 40 months
- Weight at study initiation: 2.8 - 3.0kg
- Housing: Individually in PPO cages (floor area: 2576cm^2)
- Diet (e.g. ad libitum): Altromin 2123, ad libitum
- Water (e.g. ad libitum): ad libitum, acidified with hydrochloric acid to pH 2.5 in order to prevent microbial growth
- Acclimation period: 7d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C +/- 3°C
- Humidity (%): 30% - 70%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12hrs/12hrs

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: no, but untreated eye served as control

Amount / concentration applied:

- 0.1g of the undiluted test item was placed on the conjunctival sac of one eye of the rabbit
- due to the low density of the powdery test item and the limited capacity of the conjunctival sac, it was not possible to place the large-volume amount of the test item into the eye without losses of the test item

Duration of treatment / exposure:

1hr after beginning of treatment residues of the solid were rinsed off with 0.9% NaCl

Observation period (in vivo):

1hr, 24 hrs, 48hrs, and 72hrs and 7d

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with 0.9% NaCl
- Time after start of exposure: 1hr

SCORING SYSTEM:
- according to OECD 405 and Regulation (EC) 440/2008 B.5, table 1 and table in section "Any other information on materials and methods incl. tables"

TOOL USED TO ASSESS SCORE: fluorescein and UV-light

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

animal 1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

animal 1 (with Fluorescein)

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 3

Irritation parameter:

iris score

Basis:

mean

Remarks:

animal 1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

animal 1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"; max score after 1hr: 2

Results 5

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animal 1

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 6

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

animal 2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 7

Irritation parameter:

iris score

Basis:

mean

Remarks:

animal 2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 8

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

animal 2

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 9

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animal 2

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 10

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

animal 3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 11

Irritation parameter:

iris score

Basis:

mean

Remarks:

animal 3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 12

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

animal 3

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Results 13

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animal 3

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

other: Individual data in section "Any other information on results incl. tables"

Irritant / corrosive response data:

See section "Any other information on results incl. tables"

Other effects:

See section "Any other information on results incl. tables"

Any other information on results incl. tables

TABLE: Assessment of ocular damages in the animal No. 4145 used for the initial test and in the animals No. 3813 as well as No. 3985 used for the confirmatory test

Animal No./ Body weight in kg			1 hr	24 hrs		48 hrs		72 hrs		x*		Day 7
					F		F		F		F		F
4145/ 2.9	Opacity of the cornea	Degree	0	0	1	0	0	0	0	0.00	0.33	0	0
		Area	0	0	1	0	0	0	0			0	0
	Iris		0	0		0		0		0.00		0
	Conjunctiva	Redness	2	1^a)		1		1		1.00		0
		Chemosis	1	2		1		1		1.33		0
3813/ 2.8	Opacity of the cornea	Degree	0	0	0	0	/	0	/	0.00	/	/	/
		Area	0	0	0	0	/	0	/			/	/
	Iris		0	0		0		0		0.00		/
	Conjunctiva	Redness	1	1		0		0		0.33		/
		Chemosis	1	1		0		0		0.33		/
3985/ 3.0	Opacity of the cornea	Degree	0	0	0	0	/	0	/	0.00	/	/	/
		Area	0	0	0	0	/	0	/			/	/
	Iris		0	0		0		0		0.00		/
	Conjunctiva	Redness	1	1		0		0		0.33		/
		Chemosis	1	1		0		0		0.33		/

x        mean

F         Reading after the instillation of Fluorescein.

*       Only the scores from the readings after 24 hours, 48 hours and 72 hours were included in the calculation of the individual meanvalues.

^a)       Loss of hair below the eye (probably caused by behaviour of intensive cleaning)

RESULTS

One hour after the application of the test item animal No. 4145 showed a conjunctiva with diffuse crimson colour and individual vessels not easily discernible as well as a swelling above normal. Some hyperaemic conjunctival blood vessels and a swelling above normal were observed in animals No. 3813 and No. 3985. 24 hrs after the application of the test item, animal No. 4145 showed some hyperaemic conjunctival blood vessels, an obvious swelling with partial eversion of lids and loss of hair below the eye probably caused by behaviour of intensive cleaning. Some hyperaemic conjunctival blood vessels and a swelling above normal were observed in animal No. 3813 and No. 3985. After the instillation of Fluorescein only animal No. 4145 showed scattered or diffuse areas of opacity of the cornea with details of the iris clearly visible on about one quarter of the cornea area. 48 hrs and 72 hrs after the application of the test item, some hyperaemic conjunctival blood vessels and a swelling above normal were observed in animal No. 4145. Animals No. 3813 and No. 3985 showed no ocular reactions. After the instillation of Fluorescein no changes of the cornea were revealed also in animal No. 4145. Seven days after the application of the test item, animal No. 4145 was free of any signs of eye irritation, too.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the results of the study described in this final report, EXPANDED GRAPHITE POWDER, Batch, has not to be classified as irritating to the eye according to the provisions laid down in regulation (EC) No 1272/2008.

Executive summary:

The local effect of EXPANDED GRAPHITE POWDER was investigated according to the method recommended in the OECD Guideline 405 and the council regulation (EC) No 440/2008 part B.5. Based on the critical analysis of all known data performed by the sponsor or monitor in his own responsibility the in vivo testing of the test item was considered to be justified. During an initial test, an amount of approx. 0.1 g of the undiluted test item was applied in one eye of a female albino rabbit. The eye was examined and the changes were graded according to a numerical scale after 1 hr, 24 hrs, 48 hrs and 72 hrs as well as on day 7. Changes ranging between slight and well defined were observed in the treated eye. Based on these findings two more female albino rabbits were treated with approx 0.1 g of the undiluted test item each. Likewise ocular defects were graded after 1 hr, 24 hrs, 48 hrs and 72 hrs. Both of the test animals showed slight changes concerning eyes. All effects were fully reversible within 7 days.

The following mean values, based on the results from the 24-hour-, 48-hour- and 72 hour-readings were calculated:

Animal No	Opacity of the Cornea		Iris Lesion	Redness of Conjunctiva	Oedema of Conjunctiva
4145	0.00	0.33*	0.00	1.00	0.33
3813	0.00	/*	0.00	0.33	0.33
3985	0.00	/*	0.00	0.33	0.33

*after instillation of Fluorscein

Based on the results of the study described in this final report, EXPANDED GRAPHITE POWDER has not to be classified as irritating to the eye according to the provisions laid down in regulation (EC) No 1272/2008.