Registration Dossier

Administrative data

Description of key information

Key in vivo studies for skin and eye irritation/corrosion were available for iron sulfide, demonstrating that the substance is not irritating for skin and eye. Read across was not applied for these endpoints, as it is known that other iron salts may be more water soluble and dissociate into possible irritating or corrosive constituents.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
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.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted 24 april 2002
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals 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: 2.83 – 2.94 kg
- Fasting period before study: Not provided
- 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: changed at least 1x/week). The litter collection tray (noryl waste tray) will be changed daily (except on Sundays).
- Diet (e.g. ad libitum): Rabbit feed manufactured by Pranav agro Industries Ltd., Sangli, Maharashtra, India, ad libitum.
- Water (e.g. ad libitum): 750 mL makrolon bottle fitted with stainless steel sipper tube (changed at least 1x/week), 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
- Acclimation period: One rabbit (RB9615) was acclimatized for 5 days while the other two (RB9616 and RB9617) rabbits were acclimatized for 6 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23°C
- Humidity (%): 65-67%
- Air changes (per hr): 12-15/hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: July 19, 2012 To: July 28, 2012
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
other: de-ionised water
Controls:
other:
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.2 mL of
de-ionised water
Duration of treatment / exposure:
4 hours
Observation period:
At 1, 24, 48, 72 hours, 7 and 14 days post removal of test patch.
If reversibility is seen at 72 hours, 7 days or before 14 days, the experiment is terminated.
Number of animals:
3 males
Details on study design:
TEST SITE
- Area of exposure: left side of the dorsolateral trunk
- % coverage: 6 cm2
- Type of wrap if used: cotton gauze (±6 cm : 2 x3 cm -6 ply) secured by an adhesive tape (‘Johnson –plast’ USP manufactured by Johnson & Johnson Ltd., Mumbai, India) and a crepe bandage around the torso

REMOVAL OF TEST SUBSTANCE
- Washing (if done): thoroughly with de-ionised water
- Time after start of exposure: 4 hours

SCORING SYSTEM: according to the numerical scoring system of Draize et al. (1944)
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4

Table 1. Individual Skin Reaction Scores

Rabbit

No.

Sex

Washing after (hours)

observations for skin reactions (post removal of the test patch)

Erythema score

Edema Score

Total score

C

T

C

T

C

T

RB9615*

M

4

1 hour
24 hours
48 hours
72 hours

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

RB9616

M

4

1 hour
24 hours
48 hours
72 hours

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

RB9617

M

4

1 hour
24 hours
48 hours
72 hours

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

  M: Male;          C: Control patch;                   T: Treatment patch;                          0: No Erythema/No Edema 

 * : The responses (skin reactions) were assessed immediately after the removal of the test patches at 3 minutes, 1 hour and 4 hours after application and it indicated that the test item was not irritating or corrosive.

 

 

Table 2. Mean Skin Reaction Scores

Observation (post removal of the test patch in)

 

Erythema Score

Edema Score

Total Mean Score

C

T

C

T

C

T

1 hour

 

0

0

0

0

0

0

24 hours

0

0

0

0

0

0

48 hours

0

0

0

0

0

0

72 hours

0

0

0

0

0

0

C: Control patch;                            T: Treatment patch;                         0: No Erythema/No Edema

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results, the test item, Tribotecc®-Ferrostar is “Non-irritant” to the skin of New Zealand White Rabbits under the testing conditions adopted as per Draize’s Classification Scheme.
Executive summary:

The acute dermal irritation/corrosion study in New Zealand White Rabbits was conducted to evaluate the skin irritation potential of iron sulfide (Tribotecc ®-Ferrostar). A quantity of 0.5 g of the test item was made into paste by adding 0.2 mL of de-ionised water on a cotton gauze of 6 cm2 applied to the prepared area of skin under occlusive dressing. After 4 hours contact period, the treated area was washed with de-ionised water. The study was conducted in a stepwise manner (i.e., one rabbit was dosed, followed by two additional rabbits). The degree of irritation was evaluated and scored by CLP classification method at 1, 24, 48 and 72 hours post removal of the test patch. The total mean score of skin reaction was ‘0’ at all the observation periods, indicating that the test item did not cause dermal irritation to rabbits. There were no toxic signs, pre-terminal deaths and no skin reactions observed. There were no abnormalities detected at necropsy.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
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.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted 24 April 2002
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
Annex to Commission Directive 2001/73/EC, 2004
GLP compliance:
yes (incl. certificate)
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 ??)
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
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
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0.44
Max. score:
4
Reversibility:
fully reversible within: 72 h
Irritation parameter:
other: discharge
Basis:
mean
Time point:
24/48/72 h
Score:
0.77
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
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
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
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
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
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
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.

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

 

 

 

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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

A key study for acute dermal irritation/corrosion study was conducted with iron sulfide (Tribotecc®-Ferrostar) in New Zealand White rabbits (Prakash 2012a). A quantity of 0.5 g of the test item was made into paste by adding 0.2 mL of de-ionised water on a cotton gauze of 6 cm2applied to the prepared area of skin under occlusive dressing. After 4 hours contact period, the treated area was washed with de-ionised water.

The study was conducted in a stepwise manner (i.e., one rabbit was dosed, followed by two additional rabbits).The degree of irritation was evaluated and scored by CLP classification method at 1, 24, 48 and 72 hours post removal of the test patch. The total mean score of skin reaction was ‘0’ at all the observation periods, indicating that the test item did not cause dermal irritation to rabbits. There were no toxic signs, pre-terminal deaths and no skin reactions observed. There were no abnormalities detected at necropsy.

 

A key study for acute eye irritation/corrosion was conducted with iron sulfide (Tribotecc® - Ferrostar) in New Zealand White rabbits (Praksh, 2012b). 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 and was examined at 1, 24, 48 and 72 hours post-instillation and scored. There were no toxic signs and preterminal deaths and no abnormality was detected at necropsy in any of the animals. Based on the highest total mean eye reaction Draize score of 6.67 at 24 hours post instillation and together with the persistence of the reaction the test item Tribotecc® - Ferrostar was a “Mild Irritant” according to “Modified Version of the

Kay J H and Calandra J C. (1962) System”, however based on the results of the present study the test item 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.

In vitro studies were waived because in vivo studies were availble at the time of submission.

Justification for classification or non-classification

Iron sulfide does not have to be classified for dermal and eye irritation / corrosion according the CLP No. 1272/2008 of 16 December 2008.