Iron sulphide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to GLP and valid methods, therefore it is considered relevant, reliable and adequate for classification.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 24 April 2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Annex to Commission Directive 2001/73/EC, 2004

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Iron sulphide

EC Number:

215-268-6

EC Name:

Iron sulphide

Cas Number:

1317-37-9

Molecular formula:

FeS

IUPAC Name:

sulfanylideneiron

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Toxicology, department of Safety Assessment, Advinus Therapeutics Limited, Bangalore 560 058, India
- Age at treatment: 6-7 months
- Weight at treatment: 2.60 – 2.92 kg
- Housing: Individually in rabbit cages (approx. size: L 65 x B 65 x H 45 cm) with noryl shallow cage body and facilities for pelleted food (Stainless steel feed hopper) and drinking water (750 mL makrolon bottle fitted with sipper tube). The litter collection trays (noryl waste tray) were washed daily with water (except on Sundays). The water bottle and feed hopper were changed once a week.
- Diet (e.g. ad libitum): Rabbit feed manufactured by Pranav agro Industries Ltd, Sangli, Maharashtra, India was provided ad libitum.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd., Mumbai 400 001, India provided in Makrolon bottles with stainless steel sipper tubes, ad libitum.
- Acclimation period: One rabbit was acclimatized for 5 days while the other two rabbits were acclimatized for six days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23°C
- Humidity (%): 65-66%
- Air changes (per hr): 12 - 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From:August 8, 2012 To: August 17, 2012 (August 13,2012 to August 21,2012 ??)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg (0.1g)

Duration of treatment / exposure:

single application; the lids were held together gently for about one second to prevent loss of test item.

Observation period (in vivo):

Initial single animal: 96 hours (scores only recorded through 72 hours post instillation)
Confirmatory two rabbits: up to 72 hours post application
The eyes of each rabbit were examined at 1, 24, 48 and 72 hours post-instillation and scored.

Number of animals or in vitro replicates:

3 (Initial Test: single animal test; Confirmatory Test: two additional animals)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After 24 hours contact period, the treated eye was irrigated with de-ionised water for one minute to remove the residual test item. Similarly, the control eye was also irrigated with de-ionised water.
- Time after start of exposure:24 hours

SCORING SYSTEM:
The grades of ocular reaction was assessed according to the numerical scoring system, as per Draize J H, (1977) “Dermal and Eye Toxicity Tests” In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p. 48 to 49 (Table 1)) and Classification according to “Modified Version of the Kay J H and Calandra J C. (1962) System” (Table 2).

The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of opacity). For each tissue the score was calculated as follows:
Score for conjunctivae = (A + B + C) x 2
Score for iris = D x 5
Score for cornea = (E x F) x 5
Maximum total score possible = 110
Using the numerical data obtained, a modified version of the system described by Kay J H and Calandra J C. 1962., (J. Soc. Cosmet. Chem., 13:281-289) was used to classify the ocular irritancy potential of the test item. This is achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation is calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enables the classification of the eye irritancy potential of the test item.

TOOL USED TO ASSESS SCORE: No data

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: redness

Results 2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.44

Max. score:

4

Reversibility:

fully reversible within: 72 h

Results 3

Irritation parameter:

other: discharge

Basis:

mean

Time point:

24/48/72 h

Score:

0.77

Max. score:

3

Reversibility:

fully reversible within: 72 h

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 5

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 6

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 7

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 8

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 9

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 10

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 11

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 12

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 72 h

Results 13

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 72 h

Results 14

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 72 h

Results 15

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 72 h

Results 16

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Results 17

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritant / corrosive response data:

Redness score of 1 was observed in all the three rabbits at 1, 24 & 48 hours post instillation. Chemosis score of 1 was observed in 1 rabbit at 24 & 48 hours and in the other 2 rabbits at 24 hours post instillation. In one rabbit discharge score of 2 was observed at 1 hour and 24 hours and score of 1 at 48 hours, in the other two rabbits discharge score of 2 was observed 1 hour and score of 1 at 24 & 48 hours post instillation. There were no eye reactions observed at 72 hours post instillation. There were no corneal opacity and iris reactions.

Any other information on results incl. tables

Table 1.    Individual Eye Irritation Scores

Rabbit No.	Elapsed Time	CONJUNCTIVA				IRIS		CORNEA			Total score (0-110)
		Redness (0-3) A	Chemosis (0-4) B	Discharge (0-3) C	Tissue score (0-20) (A+B+C) x 2	Pupil (0-2) D	Tissue score (0-10) Dx5	Opacity (0-4) E	Area of opacity (1-4) F	Tissue score (0-80) (E x F) x 5
RB9630	1 hour	1	0	2	6	0	0	0	NA	0	6
	24 hours	1	1	1	6	0	0	0	NA	0	6
	48 hours	1	1	1	6	0	0	0	NA	0	6
	72 hours	0	0	0	0	0	0	0	NA	0	0
Mean	24-72 h	0.66	0.66	0.66		0		0
RB9631	1 hour	1	0	2	6	0	0	0	NA	0	6
	24 hours	1	1	1	6	0	0	0	NA	0	6
	48 hours	1	0	1	4	0	0	0	NA	0	4
	72 hours	0	0	0	0	0	0	0	NA	0	0
Mean	24-72 h	0.66	0.33	0.66		0		0
RB9632	1 hour	1	0	2	6	0	0	0	NA	0	6
	24 hours	1	1	2	8	0	0	0	NA	0	8
	48 hours	1	0	1	4	0	0	0	NA	0	4
	72 hours	0	0	0	0	0	0	0	NA	0	0
Mean	24-72 h	0.66	0.33	1		0		0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the highest total mean eye reaction score of “6.67” at 24 hours post instillation and together with the persistence of the reaction the test item Tribotecc® - Ferrostar is a “Mild Irritant” according to “Modified Version of the Kay J H and Calandra J C. (1962) System”.
Based on the results of the present study the test item Tribotecc® - Ferrostar however does not need to be classified as per Regulation (EC) No 1272/2008 of the European Parliament and of the council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006.

Executive summary:

The acute eye irritation / corrosion study in New Zealand White rabbits was conducted to evaluate the eye irritation potential of iron sulfide (Tribotecc® - Ferrostar). On test day one, a quantity of 100 mg (0.1g) of test item was instilled into the conjunctival sac of the left eye of the animal after gently pulling the lower lid away from the eyeball. The lids were then held together gently for about one second to prevent loss of test item. The right eye remained untreated and served as the reference control. All the rabbits (one rabbit for initial test and two rabbits for confirmatory test) were treated in a similar manner. The eyes of each rabbit were examined at 1, 24, 48 and 72 hours post-instillation and scored. The grades of ocular reaction were assessed as per Draize. There were no toxic signs and pre-terminal deaths and no abnormality was detected at necropsy in any of the animals. Based on the highest total mean eye reaction score of 6.67 at 24 hours post instillation and together with the persistence of the reaction the test item Tribotecc® - Ferrostar is a “Mild Irritant” according to “Modified Version of the Kay J H and Calandra J C. (1962) System”. Based on the results of the present study the test item Tribotecc® - Ferrostar however does not need to be classified as per Regulation (EC) No 1272/2008 of the European Parliament and of the council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006.