Tin disulphide

Endpoint summary

Description of key information

Both in vitro and in vivo studies were performed in a tiered testing approach, demonstrating that Tin disulfide is not corrosive or irritating to skin and eye. The same approach was done for read-across substance

Tin sulfide, which also was negative for skin and eye corrosion and irritation.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

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 standard methods under GLP as a final step of skin irritation testing. The study is reliable, relevant, and adequate for classifcation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products Inc., Denver, PA
- Age at study initiation: 5-6 months
- Weight at study initiation: 2.8 - 3.3 kg
- Fasting period before study: Not provided
- Housing: Individually in suspended cages. Absorbent paper bedding was placed beneath the cages and changed at least three times per week.
- Diet (e.g. ad libitum): Fresh PMI Rabbit Chow (Diet #5321) was provided daily
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Controlled
- Humidity (%): Not provided
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: distilled water

Controls:

not required

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): 0.3 mL

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: 6cm2 (2 x 3 cm gauze patch)
- Type of wrap if used: porous dressing (semi-occlusive) and porous, non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): gently washing with distilled water at the end of the exposure period
- Time after start of exposure: 4 hours

SCORING SYSTEM: Erythema and edema were scored according to the numerical Draize technique below. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction. Additional signs were described.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24-72 h

Score:

0.55

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24-72 h

Score:

0.55

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24-72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: no edema

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: no oedema

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: no oedema

Table 2. Dermal and Systemic observations

Time after patch removal		Rabbit H4909 male	Rabbit H4933 male	Rabbit H4936 male	Mean Scores
1 Hour	Erythema	1	1	0
	Edema	0	0	0
Systemic Observations		A	A	A
24 Hour	Erythema	1s	1	1	1.00
	Edema	0	0	0	0.00
Systemic Observations		A	A	A
48 Hour	Erythema	1s	0	0f	0.33
	Edema	0	0	0	0.00
Systemic Observations		A	A	A
72 Hour	Erythema	1	0	0f	0.33
	Edema	0	0	0	0.00
Systemic Observations		A	A	A
Day 7	Erythema	0	0	0
	Edema	0	0	0
Systemic Observations		A	A	A
			Sum of Mean Scores =		1.66
Modified Primary Irritation Index (PII)			=   Sum of Mean Scores/3 =		0.55
A=appeared normal			f= flaking skin		s= shiny areas

Interpretation of results:

GHS criteria not met

Conclusions:

The Modified Primary irritation Index is 0.55. Tin disulfide is a dermal irritant.

Executive summary:

Three healthy New Zealand White rabbits (males) were dosed dermally with Tin disulfide. The test article (0.56 g) was applied dermally to one intact site per rabbit and wrapped with a piece of porous dressing (semi-occlusive) large enough to cover all dose sites with at least 5 cm square to spare on all sides of the gauze patch. The test article was kept in contact with the skin for 4 hours at which time the wrappings were removed. Erythema and edema were scored at 1, 24, 48 and 72 hours and on Day 7 following patch removal. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction at these time periods. Animals were observed for toxicological and pharmacological effects at each dermal observation period and observed for mortality daily. Body weights were recorded pretest, at 72 hours and at termination. At 1 hour post exposure, erythema was absent to very slight and edema was absent. At 24 hours erythema was very slight, edema was absent and shiny areas were observed on one animal. By 48 hours, erythema was absent to very slight and edema was absent with shiny areas and flaking skin observed. Erythema remained absent to very slight, edema was absent and flaking skin persisted at 72 hours. Both erythema and edema were absent by Day 7. No abnormal physical signs were observed. Two animals had net body weight gain by study termination, although one of the animals lost weight between 72 hours and Day 7. The remaining animal lost weight by study termination. The Modified Primary Irritation Index was 0.55. Tin disulfide (CAS 1315-01-1; EINECS: 215- 252-9), Tin disulphide; Trade name: Tribotecc® - SNS2 Grade A, Batch# E 10320 is not a Category 2 irritant according to Regulation (EC) No. 1272/2008, Section 3.2.2.7.

Reference 2

Endpoint:

skin irritation: in vitro / ex 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 standard methods under GLP as a second (in vitro) step of skin irritation testing. The study is reliable and relevant, but as a stand-alone study with negative result, not adequate for classifcation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: not specified

Source strain:

not specified

Vehicle:

other: PBS was used to improve the contact between powder and tissue.

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: MatTek EpiDermTM Skin Irritation Test
- Tissue batch number(s): Lot 12907 Kit D
- Production date: Not specified
- Shipping date: Not specified
- Delivery date: 13 Dec 2011
- Date of initiation of testing: 1st exposure to test substance 14 December 2011

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: The exposure period for the test item and controls was 60 minutes. The dosed tissues were placed in an incubator at 37°C ± 1°C, 5% ± 1% CO2 for 35 ± 1 minutes, then returned to the sterile hood for the remainder of the 60-minute exposure period.
- Temperature of post-treatment incubation (if applicable): No temperature data of post-treatment incubation.
After dosing and incubation, the tissues were rinsed with PBS, blotted to remove the test substance and dry the tissue, and transferred to fresh medium. The rinsed EpiDerm TM tissues were returned to the incubator for 24 ± 2 hours. Medium was changed at 24 ± 2 hrs. Tissues were returned to the
incubator for an additional 18 ± 2 hours.
At the end of the incubation period, each EpiDerm™ tissue was rinsed with phosphate buffered saline (PBS) and transferred to a 24-well plate containing 300 μL of MTT solution (1 mg/mL MTT in DMEM). The tissues were then returned to the incubator for a three-hour MTT incubation period.
Following the MTT incubation period, each EpiDerm™ tissue was rinsed with PBS and then treated with 2.0 mL of extractant solution (isopropanol) per well for at least 2 hours, with shaking, at room temperature. Two aliquots of the extracted MTT formazan were measured at 540 nm using a plate
reader (μQuant Plate Reader, Bio-Tek instruments, Winooski, VT).

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 3 washing steps with PBS, no data on volume used.
After dosing and incubation, the tissues were rinsed with PBS, blotted to remove the test substance and dry the tissue, and transferred to fresh medium. The rinsed EpiDerm TM tissues were returned to the incubator for 24 ± 2 hours. Medium was changed at 24 ± 2 hrs. Tissues were returned to the
incubator for an additional 18 ± 2 hours.
At the end of the incubation period, each EpiDerm™ tissue was rinsed with phosphate buffered saline (PBS) and transferred to a 24-well plate containing 300 μL of MTT solution (1 mg/mL MTT in DMEM). The tissues were then returned to the incubator for a three-hour MTT incubation period.
Following the MTT incubation period, each EpiDerm™ tissue was rinsed with PBS and then treated with 2.0 mL of extractant solution (isopropanol) per well for at least 2 hours, with shaking, at room temperature. Two aliquots of the extracted MTT formazan were measured at 540 nm using a plate reader (μQuant Plate Reader, Bio-Tek instruments, Winooski, VT).
- Observable damage in the tissue due to washing: Not specified
- Modifications to validated SOP: There were no amendments to the protocol. The test article char
acterization was not conducted according to the Good Laboratory Practices. This is not expected to
have any impact on the study.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL MTT in DMEM
- Incubation time: three-hour MTT incubation period
- Spectrophotometer: a plate reader (μQuant Plate Reader, Bio-Tek instruments, Winooski, VT)
- Wavelength: 540 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
Not specified

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
No direct MTT interference was observed. The test article did not reduce MTT and the assay cont
inued as per the protocol.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL

PREDICTION: 6 tests (3 replicates with each two aliquots)

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritant to skin if the mean tissue viability after 60 minutes
exposure is smaller than or equal to 50%,
- The test substance is considered to be non-irritant to skin if the mean tissue viability after 60 minutes
exposure is greater than 50%

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg of the test article was added into the Milllicell
atop the tissue

VEHICLE
- Amount(s) applied (volume or weight with unit): The EpiDermTM tissue was moistened with 25 μL
PBS before dosing
- Lot/batch no. (if required): Lot#AWD10434)

NEGATIVE CONTROL: PBS
- Amount(s) applied (volume or weight): not specified
- Concentration (if solution):

POSITIVE CONTROL: 5% SDS
- Amount(s) applied (volume or weight): not specified
- Concentration (if solution): 5%

Duration of treatment / exposure:

60 minutes

Duration of post-treatment incubation (if applicable):

42 ± 2 hours . After dosing and incubation, the tissues were rinsed with PBS, blotted to remove the test substance and dry the tissue, and transferred to fresh medium. The rinsed EpiDerm TM tissues were returned to the incubator for 24 ± 2 hours. Medium was changed at 24 ± 2 hrs. Tissues were returned to the incubator for an additional 18 ± 2 hours.

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean

Run / experiment:

60 minutes

Value:

101.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: Not specified.
- Direct-MTT reduction: No direct MTT interference was observed. The test article did not reduce M
TT.
- Colour interference with MTT: Not specified.

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes. The mean OD540 of the Negative Control tissues
is ≥ 1.0 and ≤ 2.5 (2.058).
- Acceptance criteria met for positive control: Yes. The mean viability of the Positive Control tissues, e
xpressed as percentage of the negative control tissues is ≤ 20% (9.6%)
- Acceptance criteria met for variability between replicate measurements: Yes. The standard deviation
(SD) calculated from individual percent tissue viabilities of the three identically-treated replicates is <
18% (SD 0.46-2.66).

Table 1. Results

Test Article Identity	Tissue No.	Raw data		Blank corrected data		Mean of Ali-quots	% Viabi-lity	OD		Viabilities		CV%	Classification
		Aliq.1	Aliq.2	Aliq.1	Aliq.2			Mean	SD	Mean	SD
Tin disulfide Batch E10320	1	2.088	2.066	2.046	2.024	2.035	98.9	2.084	0.055	101.2	2.66	2.63	Non-Irritant
	2	2.185	2.185	2.143	2.143	2.143	104.1
	3	2.094	2.137	2.052	2.095	2.073	100.7
PBS (negative control)	1	2.096	2.094	2.054	2.052	2.053	99.7	2.058	0.047	100.0	2.28	2.28	Non-Irritant
	2	2.059	2.054	2.017	2.012	2.014	97.9
	3	2.141	2.159	2.099	2.117	2.108	102.4
5% Sodium dodecyl sulfate  (positive control)	1	0.251	0.249	0.209	0.207	0.208	10.1	0.198	0.010	9.6	0.46	4.81	Irritant
	2	0.240	0.239	0.198	0.197	0.197	9.6
	3	0.229	0.233	0.187	0.191	0.189	9.2

 

Interpretation of results:

GHS criteria not met

Conclusions:

Mean Tissue Viability of tin disulfide was 101.2%, accordingly tin disulfide was predicted non-irritant.

Executive summary:

MatTek EpiDerm (TM) tissue samples were treated in triplicate with the test article, negative control (Phosphate buffered saline) and positive control (5% Sodium dodecyl sulfate) for 60 minutes, followed by a 42 hour recovery period. Following treatment and subsequent incubation time, the viability of the tissues was determined using Methyl thiazole tetrazolioum (MTT) uptake and reduction. The absorbance of each sample was measured at 540 nm. The viability was then expressed as a percent of control values. If the mean tissue viability was ≤50%, the test material was classified as an irritant; if the mean tissue viability was >50%, the test material was classified as a non-irritant.

The test article tin disulfide was non-irritant in the MatTek EpiDerm (TM) Skin Irritation Test, since its mean tissue viability was 101.2%.

Reference 3

Endpoint:

skin irritation: in vitro / ex 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 standard methods under GLP as a first (in vitro) step of skin corrosion and irritation testing. The study is reliable and relevant, but as a stand-alone study with negative result, not adequate for classifcation.

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 431

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: not specified

Source strain:

not specified

Details on animal used as source of test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: MatTek EpiDermTM Skin Corrosivity Test
- Tissue batch number(s): Lot 16216 Kit H
- Production date: Not specified
- Shipping date: Not specified
- Delivery date: 13 December 2011
- Date of initiation of testing: 1st exposure to test substance 14 December 2011

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: No temperature data during exposure.
- Temperature of post-treatment incubation (if applicable): No temperature data of post-treatment incubation.
At the end of the exposure period, each EpiDerm TM tissue was rinsed with phosphate buffered saline (PBS) and transferred to a 24-well plate containing 300 μL of MTT solution (1 mg/mL MTT in
DMEM). The tissues were then returned to the incubator for a three-hour MTT incubation period. Following the MTT incubation period, each EpiDerm™ tissue was rinsed with PBS and then treated
overnight with 2.0 mL of extractant solution (isopropanol) per well. The absorbency of an aliquot of the extracted MTT formazan was measured at 540 nm using a microplate reader (μQuant Plate Reader,
Bio-Tek Instruments, Winooski, VT).

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 2 washing steps with PBS, no data on volume used.
At the end of the exposure period, each EpiDerm TM tissue was rinsed with phosphate buffered saline (PBS) and transferred to a 24-well plate containing 300 μL of MTT solution (1 mg/mL MTT in
DMEM). The tissues were then returned to the incubator for a three-hour MTT incubation period. Following the MTT incubation period, each EpiDerm™ tissue was rinsed with PBS and then treated
overnight with 2.0 mL of extractant solution (isopropanol) per well. The absorbency of an aliquot of the extracted MTT formazan was measured at 540 nm using a microplate reader.
- Observable damage in the tissue due to washing: Not specified
- Modifications to validated SOP: There were no amendments to the protocol. The test article characterization was not conducted according to the Good Laboratory Practices. This is not expected to
have any impact on the study.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL MTT in DMEM
- Incubation time: three-hour MTT incubation period
- Spectrophotometer: a microplate reader
- Wavelength: 540 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
Not specified

NUMBER OF REPLICATE TISSUES: 2

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
No direct MTT interference was observed. The test article did not reduce MTT and the assay continued as per the protocol.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 12 tests (2 replicates with each 3 aliquots at 2 time points)

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the
viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Approximately 25 mg of the test article plus 25 μL tissue culture water were applied to the top of each EpiDerm tissue.

Duration of treatment / exposure:

3 and 60 minutes

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean

Run / experiment:

3 minutes

Value:

99.4

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean

Run / experiment:

60 minutes

Value:

96.7

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: Not specified.
- Direct-MTT reduction: No direct MTT interference was observed. The test article did not reduce MTT.
- Colour interference with MTT: Not specified.

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes. The mean OD of the two Negative Control tissues is ≥ 0.8 (1.65 at 3 minutes and 1.63 at 60 minutes).
- Acceptance criteria met for positive control: Yes. The mean relative tissue viability of the Positive Control tissues at the 60-minute time point is < 15% (11.52%).
- Acceptance criteria met for variability between replicate measurements: Yes. The difference between two identically treated tissues is no greater than 30%: Viability differences between the
two identically treated tissues in all samples and controls at 3 minutes were 1.2% to 9.6%. Viability differences between the two identically treated tissues at 60 minutes were 0.9% to 9.2%.

Table 1. Tin disulfide (batch# E 10320); dose 25 mg plus 25 µL TCH₂O; conc. Neat

Time (min)	OD1	OD2	OD3	Mean (OD)	SD	Viability %	Error %
3.0	1.673	1.636	1.636	1.648	0.021	100.0	1.3
	1.611	1.631	1.642	1.628	0.016	98.8	1.0
neg control				1.648		100.0
60.0	1.634	1.627	1.629	1.630	0.004	99.7	0.2
	1.525	1.529	1.543	1.532	0.009	93.7	0.6
neg control				1.635		100.0
Mean % Viability at 3 min.:						99.4
Mean % Viability at 60 min.:						96.7
Predicted Corrosivity:			Non-Corrosive

Interpretation of results:

GHS criteria not met

Conclusions:

Predicted corrosivity of tin disulfide in the EpiDermTM Skin Irritation test was non-corrosive. Mean viability of tin disulfide was 99.4% (3 min) and 96.7% (60 min).

Executive summary:

MatTek EpiDerm(TM) tissue samples were treated in duplicate with tin disulfide, negative control (tissue culture water) and positive control (Potassium Hydroxide, 8.0N) for 3 minutes and 60 minutes. Following treatment, the viability of the tissues was determined using Methyl thiazole tetrazolium (MTT) uptake and conversion, and the absorbance of each sample was measured at 540 nm. The viability was then expressed as a percent of control values. The percent viability was used to determine corrosivity potential. Mean viability of tin disulfide was 99.4% (3 min) and 96.7% (60 min). Predicted corrosivity of tin disulfide in the EpiDerm (TM) Skin Irritation test was non-corrosive.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye 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 standard methods under GLP as a final step of skin irritation testing. The study is reliable, relevant, and adequate for classifcation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Covance Research products, Inc., Denver, PA
- Age at study initiation: ± 5 months
- Weight at study initiation: 3.0-3.1 kg
- Fasting period before study: Not provided
- Housing: Individually in suspended cages. Absorbent paper bedding was placed beneath the cages and changed at least three times per week.
- Diet (e.g. ad libitum): Fresh PMI Rabbit Chow (#5321) was provided daily.
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Temperature controlled
- Humidity (%): Not provided
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From 2012-01-31 To 2012 02 02

Vehicle:

unchanged (no vehicle)

Controls:

other: non treated eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL equivalent 94.4 mg
- Concentration (if solution):

Duration of treatment / exposure:

single application, the lids were held close for approximately 1 second after instillation

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 (males)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: Draize technique

TOOL USED TO ASSESS SCORE: Mini-Maglite® flashlight

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0.67

Max. score:

110

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.11

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: 2 animals with eye redness

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

Max. score:

3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Interpretation of results:

GHS criteria not met

Conclusions:

Tin disulfide is not an ocular irritant according to CLP & GHS Guidelines.

Executive summary:

Three healthy New Zealand White rabbits (male), free from evidence of ocular irritation and corneal abnormalities were dosed with tin disulfide. The test article (0.1 mL equivalent (94.4 mg)) was placed into the conjunctival sac of one eye of each rabbit. The contralateral eye served as a control. The eyes were examined pretest and scored by the Draize technique at 1, 24, 48 and 72 hours postdose.

There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation of redness and or discharge, was noted in two out of three eyes, one eye cleared by 24 hours and the other eye by 48 hours. The control eyes appeared normal at all observation periods. There were no abnormal physical signs noted during the observation period. One animal gained body weight and the other two animals remained the same.

Reference 2

Endpoint:

eye irritation: in vitro / ex 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 standard methods under GLP as a first (in vitro) step of eye damage and irritation testing. The study is reliable and relevant, but as a stand-alone study with negative result, not adequate for classifcation.

Qualifier:

according to guideline

Guideline:

other: INVITTOX.1992. Protocol No. 47: HET-CAM Test

GLP compliance:

yes (incl. QA statement)

Species:

other: not applicable; fertile, white Leghorn eggs

Strain:

other: not applicable; fertile, white Leghorn eggs

Details on test animals or tissues and environmental conditions:

Fertile, White Leghorn eggs (twenty four) received from Moyer's Chicks, Quakertown, PA were selected for use from a larger group and incubated on 02 Dec 2011. The eggs were kept in incubators at 99 ± 2°F for 10 days.
Pre-dose Procedures
The eggs were marked on one side with an "X" and on the other side with an "0", and placed horizontally in the incubator trays. The eggs were rotated once daily during the first 9 days of incubation to ensure even atmospheric exposure.
On day 9 of incubation, the eggs were rotated and turned up in the incubator with the large end upwards containing the air sac to facilitate access to the CAM.
On day 10 of development, the eggs were removed from the incubator and candled to determine the viability of the embryo. A rectangular window was removed from the shell directly over the air cell using a rotating Dremel® drill with a diamond wheel bit. The egg membrane was carefully moistened with 2-3 mL 0.9% saline and returned to the incubator. Eggs were examined for any abnormalities. All abnormal eggs were discarded.

Vehicle:

other: olive oil, Lot" L 1234R H0803 by Bertolli

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.3mL of 10% mixture of test article
- Concentration (if solution): 10% test article in olive oil

VEHICLE: Olive Oil
- Lot/batch no. (if required):Lot# L 1234R H0803

Duration of treatment / exposure:

5 minutes

Observation period (in vivo):

5 minutes during exposure

Number of animals or in vitro replicates:

6 eggs per group (test item, olive oil, 0.1N NaOH, 1% SDS)

Details on study design:

The chorioallantoic membrane (CAM) of 24 White Leghorn eggs, incubated for 10 days, were dosed with 0.3 mL of the test substance as listed in study report. The eggs were observed continuously for 5 minutes immediately following dosing for the appearance of lysis (sec L), hemorrhage (sec h) and/ or coagulation (sec C). In addition, the eggs were scored for severity at 1 and 5 minutes postdose. The irritating potential of the test article was classified based on the irritation score (IS) and the threshold concentration (TH).

Irritation parameter:

in vitro irritation score

Run / experiment:

Test item

Value:

0

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Table 1. Results

	Observations			Severity
	Hemorrhage (H)	Vessel Lysis (L)	Coagulation (C)	Score	Score
Egg #	(sec)	(sec)	(sec)	@1min	@5min	IS Score	Mean	SD
Positive Control: 0.1N NaOH
1	38	23	62	2	3	18.04	17.72	0.274
2	44	17	84	2	3	17.42
3	45	19	80	2	3	17.48
4	36	18	71	2	3	17.92
5	20	19	79	2	3	17.92
6	26	40	73	2	3	17.51
Positive Control: 1% SDS
7	53	32	301	1	2	10.41	10.20	0.200
8	56	29	301	1	2	10.43
9	63	35	301	1	2	10.17
10	67	39	301	1	2	10.01
11	70	40	301	1	2	9.94
12	82	20	301	1	2	10.21
Vehicle Control: Olive oil
13	301	301	301	0	0	0.00	0.00	0.000
14	301	301	301	0	0	0.00
15	301	301	301	0	0	0.00
16	301	301	301	0	0	0.00
17	301	301	301	0	0	0.00
18	301	301	301	0	0	0.00
10% Tin Disulfide, TriboteccSNS2 Grade A, Lot/batch# E10320
19	301	301	301	0	0	0.00	0.00	0.000
20	301	301	301	0	0	0.00
21	301	301	301	0	0	0.00
22	301	301	301	0	0	0.00
23	301	301	301	0	0	0.00
24	301	301	301	0	0	0.00

   

The mean IS for 0.1 N NaOH and 1% SDS (positive controls) were 17.72 and 10.20, respectively. The vehicle control, olive oil, had no adverse effects on the CAM. At 10% (w/v) in olive oil, the IS of the test article was 0. The threshold concentration for this test article was greater than 10%.
Based on the threshold concentration of greater than 10% and the IS 10% of 0, the irritating potential of the test article, was determined as none to slight.   

                                                                                            

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the threshold concentration of greater than 10% and the IS 10% of 0, the irritating potential of the test article, was determined as none to slight.

Executive summary:

This study was aimed to determine the potential irritancy using an alternative to the Draize methodology : the method described in INVITTOX.1992. Protocol No. 47: HET-CAM Test. The chorioallantoic membrane (CAM) of 24 White Leghorn eggs, incubated for 10 days, were dosed with 0.3 mL of 10% test substance formulation (6 eggs with 0.1N NaOH; 6 eggs with 1% Dodecyl Sulfate sodium; 6 eggs with olive oil; 6 eggs with 10% tin disulfide in olive oil). The eggs were observed continuously for 5 minutes immediately following dosing for the appearance of lysis (sec L), hemorrhage (sec H) and/or coagulation (sec C). In addition, the eggs were scored for severity at 1 and 5 minutes postdose. The irritating potential of the test article was classified based on the irritation score (IS) and the threshold concentration (TH).

The mean IS for 0.1 N NaOH and 1% SDS (positive controls) were 17.72 and 10.20, respectively. The vehicle control, olive oil, had no adverse effects on the CAM. At 10% (w/v) in olive oil, the IS of the test article was 0. The threshold concentration for this test article was greater than 10%. Based on the threshold concentration of greater than 10% and the IS 10% of 0, the irritating potential of the test article, was determined as none to slight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro studies to predict skin corrosion and irritation of tin disulfide were performed by means of MatTek EpiDerm(TM) according to OECD 431 (Piehl, 2012a) and OECD 439 method (Piehl, 2012b). In the first study, mean tissue viability was 99.4% (3 min) and 96.7% (60 min); predicted corrosivity of tin disulfide was non-corrosive. In the second study, mean tissue viability was 101.2%, therefore predicted irritation potential was non-irritant. These studies, with negative outcome, were not enough for classification, therefore they were considered to be supportive. As a final step, a key in vivo study for skin irritation was performed in New Zealand White rabbits with tin disulfide by semi-occlusive dermal application according to OECD 404 method (Yasso, 2012). Erythema and edema were scored at 1, 24, 48 and 72 hours and on Day 7 following patch removal. At 1 hour post exposure, erythema was absent to very slight and edema was absent. At 24 hours erythema was very slight, edema was absent and shiny areas were observed in one animal. By 48 hours, erythema was absent to very slight and edema was absent with shiny areas and flaking skin observed. Erythema remained absent to very slight, edema was absent and flaking skin persisted at 72 hours. Both erythema and edema were absent by Day 7. No abnormal physical signs were observed.Two animals had net body weight gain by study termination, although one of the animals lost weight between 72 hours and Day 7. The remaining animal lost weight by study termination. The Modified Primary Irritation Index was 0.55. In conclusion, tin disulfide is not a Category 2 irritant according to CLP Regulation. Supporting information was also available from tin sulfide as read across substance. MatTek EpiDerm(TM) testing according to OECD 431 showed a mean tissue viability of 101.3% (3 min.) and 107.8% (60 min.), therefore predicted corrosivity of tin sulfide was non-corrosive (Piehl, 2010a). Dermal irritation potential tested in MatTek Epiderm(TM) according to OECD 439 showed a mean tissue viability of 101.5%, therefore predicted irritation potential of tin sulfide was non-irritant (Piehl, 2010b). Finally, in vivo testing for skin irritation in rabbits according to OECD method 404 demonstrated that there was no erythema or edema observed after 4 hours semi-occlusive application. Test article stained the dose sites at all observation periods, but there were no abnormal physical signs and body weights were normal. Tin disulfide and Tin sulfide showed a comparable tolerance profile for this endpoint.

 

An in vitro study to predict eye damage and irritation of tin disulfide was performed by means of HETCAM test according to Invittox method No. 47 (Cerven, 2012). The chorioallantoic membrane (CAM) of White Leghorn eggs, incubated for 10 days, was dosed with 0.3 mL test substance at 10% (w/v) in olive oil, leading to an irritation score (IS) of 0. Based on a threshold concentration of >10% and the IS 10% of 0, the irritating potential of the test article, was determined as none to slight. This study, with negative outcome, was not enough for classification, therefore it was considered to be supportive. Next, a key in vivo study according to Draize method was performed with tin disulfide in three New Zealand White rabbits (Blair, 2012). The test article (0.1 mL equivalent (94.4 mg)) was placed into the conjunctival sac of one eye of each rabbit. The eyes were examined pretest and scored by the Draize technique at 1, 24, 48 and 72 hours postdose. There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation of redness and or discharge, was noted in two out of three eyes, one eye cleared by 24 hours and the other eye by 48 hours. The control eyes appeared normal at all observation periods. There were no abnormal physical signs noted during the observation period. One animal gained body weight and the other two animals remained the same. Supporting information was also available from tin sulfide as read across substance. HETCAM testing according to Invittox showed an irritation score of 0 (Cerven, 2010), whereas in vivo testing for eye irritation in rabbits according to OECD method 405 demonstrated that there was no eye irritation (Blair, 2010). Tin disulfide and Tin sulfide showed a comparable tolerance profile for this endpoint.

 

Justification for classification or non-classification

No classification and labelling is needed for skin and eye corrosion or irritation of tin disulfide according to EU labelling regulations Commission Directive 93/21/EEC and CLP regulation (No. 1272/2008 of 16 December 2008).