Registration Dossier

Administrative data

Description of key information

In an in vitro EpiSkin assay, conducted in accordance with OECD Test Guideline 439 and to GLP, tetraammineplatinum dinitrate was considered to be non-irritating to skin (Kiss, 2012).

 

In an OECD Test Guideline 405 in vivo eye irritation study, conducted to GLP, tetraammineplatinum dinitrate was not irritating following instillation of the test material (0.1 ml) into the right eye of 3 rabbits (Pooles, 2012). Further, in an in vitro BCOP assay, conducted in accordance with OECD Test Guideline 437, tetraammineplatinum dinitrate did not induce a response indicative of a corrosive or severe irritant potential (Warren, 2012).

 

No relevant respiratory tract irritation data were identified.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22-24 February 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Good-quality, well-documented study in accordance with OECD guideline.
Qualifier:
according to guideline
Guideline:
other: OECD Guidelines for Testing of Chemicals, Section 4, No. 439, “In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method” adopted 22 July 2010
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EpiSkin™ SOP, Version 1.8 (February 2009), ECVAM Skin Irritation Validation Study: Validation of the EpiSkin™ test method 15 min - 42 hours for the prediction of acute skin irritation of chemicals. Available at: [http://ecvam.jrc.ec.europa.eu]
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: IN VITRO SKIN IRRITATION: RECONSTRUCTED HUMAN EPIDERMIS MODEL TEST in relation to Regulation (EC) No 440/2008 (as amended) and Regulation (EC) No 1907/2006 on REACH (Annex III, B.46).
Deviations:
no
Principles of method if other than guideline:
The test is designed to predict and classify the skin irritant potential of chemicals according to chemical safety regulations, using the reconstructed human epidermis model EPISKIN-SM and parameters related to skin irritation.
EPISKIN-SM is a three-dimensional human skin model comprising a reconstructed epidermis with a functional stratum corneum. Its use for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability. Cell viability determination is based on cellular mitochondrial dehydrogenase activity, measured by MTT reduction and conversion into a blue formazan salt that is quantitatively measured after extraction from tissues. The reduction of cell viability in treated tissues is compared to negative controls and expressed as a %. The % reduction in viability is used to predict the irritation potential.

The EPISKIN-SM has been found scientifically valid for reliably predicting no label and R38 (irritant) substances in respect to the previous EU classification scheme and has been confirmed in April 2009 by ESAC for use under the UN GHS system as "applicable to all authorities". It is approved by international regulatory agencies as a replacement for the identification of irritants/corrosives in the in vivo rabbit skin assay(OECD 404).
GLP compliance:
yes (incl. QA statement)
Species:
human
Strain:
other: Reconstructed human epidermis model (see details below)
Details on test animals or test system and environmental conditions:
EPISKIN-SM (Source: SkinEthic, France, Batch No.:12-EKIN-008, Expiry date: 27 February 2012) is a three-dimensional human epidermis model. Adult human-derived epidermal keratinocytes are seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. A highly differentiated and stratified epidermis model is obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.
Type of coverage:
other: 20 ul was applied evenly to the epidermal surface
Preparation of test site:
other: in vitro cell culture
Vehicle:
unchanged (no vehicle)
Controls:
other: negative control skin unit tested in triplicate
Amount / concentration applied:
20 ul to each of three test skin units.
20 µl PBS (phosphate buffered saline) was added to each of the three negative control skin units and 20 µl SDS (sodium dodecyl sulfate, 5% aqueous solution) was added to each of the three positive control skin units.
Duration of treatment / exposure:
Exposure for 15 minutes (± 0.5 min) at room temperature (20-37°C).
Then incubated with fresh “maintenance medium” for 42 hours (± 1h) at 37°C.
Observation period:
Not applicable to this test system
Number of animals:
Not applicable to this test system
Details on study design:
EPISKIN-SM assay plate contained reconstructed epidermis units (area: 0.38 cm2); each was attached to the base of a tissue culture vessel and maintained on nutritive agar.

After test substance exposure and subsequent incubation, preparations for cell viability determination were: incubation with MTT solution (at 37 degrees C for 3 hours) followed by incubation with acidified isopropanol for formazan extraction (around two hours at room temperature with gentle agitation).

For cell viability measurements, the OD (Absorbance / Optical Density) of the samples in a spectrophotometer was read at 540 nm using acidified isopropanol solution blank (6×200 µL). (The validity of the microplate reader was verified with a standard verification plate daily before use. The standard plate was calibrated yearly by the manufacturer.)

For each treated tissue, OD was calculated and the tissue viability was expressed as a % relative to negative control. No adjustment to OD results was necessary because the test substance was colourless.

Criteria for classification as irritant/non-irritant: If the resulting mean relative viability (as adjusted for intrinsic colour) is less than or equal to 50% of the negative control, the test substance is considered to be irritant to skin.

Irritation / corrosion parameter:
other: other:
Value:
95
Remarks on result:
other:
Remarks:
Basis: mean Cell/tissue viability. Time point: 42 hours. Reversibility: other: Not applicable. Remarks: Score is a percentage (%) of negative control. Mean relative viability <=50% the test substance is considered to be irritant to skin.. (migrated information)
Irritant / corrosive response data:
Mean cell viability was 95% of the negative control (range 85%-100%), see attached table.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an in vitro reconstructed human epidermis (EpiSkin) assay, conducted in accordance with OECD Test Guideline 439 and to GLP, tetraammineplatinum dinitrate was considered to be non-irritating to skin.
Executive summary:

Tetraammineplatinum dinitrate was tested for skin irritation potential i n an in vitro reconstructed human epidermis model (EPISKIN assay) conducted in accordance with OECD Test Guideline 439, and to GLP.

 

EPISKIN is a three-dimensional human skin model comprising a reconstructed epidermis with a functional stratum corneum. Its use for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability. Cell viability determination is based on cellular mitochondrial dehydrogenase activity, measured by MTT reduction and conversion into a blue formazan salt that is quantitatively measured after extraction from tissues. The reduction of cell viability in treated tissues is compared to negative controls and expressed as a %. If the resulting mean relative viability (as adjusted for intrinsic colour) is less than or equal to 50% of the negative control, the test substance is considered to be irritant to skin.

 

Following a 15-minute exposure to the test substance, the test system skin cell viability was calculated to be greater than 50% (the average was 95% and the range was 85-100%), and it was therefore considered to be non-irritating to skin.

 

Under the conditions of this assay, tetraammineplatinum dinitrate does not require classification for skin irritation according to EU CLP criteria (EC 1272/2008).

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
Study period:
17 to 27 July 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline study to GLP
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source:Harlan Laboratories UK Ltd, Leicestershire, UK
- Age at study initiation: 12 to 20 weeks
- Weight at study initiation: 2.64 to 3.02 kg
- Housing: Individually housed in suspended cages
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23oC
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 to 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml

VEHICLE
Not applicable
Duration of treatment / exposure:
Eyes were held closed for 1 second after instillation
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
3 (Initially one animal, after consideration of the ocular response two additional animals were treated).
Details on study design:
REMOVAL OF TEST SUBSTANCE
Eyes were unwashed after treatment

SCORING SYSTEM: Ocular irritation evaluated through scoring of the conjunctivae, iris and cornea according to the Draize numerical scale.

TOOL USED TO ASSESS SCORE: Standard opthalmoscope as light source
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
110
Reversibility:
other: not applicable since no effects seen
Irritation parameter:
overall irritation score
Basis:
animal #2
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
110
Reversibility:
other: not applicable since no effects seen
Irritation parameter:
overall irritation score
Basis:
animal #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
110
Reversibility:
other: not applicable since no effects seen
Irritant / corrosive response data:
Minimal transient conjuctival redness was noted in one treated eye one hour after treatment
Other effects:
No signs of toxicity were noted during the study and all animals showed expected bodyweight gain.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an OECD Test Guideline 405 eye irritation study, conducted to GLP, tetraammineplatinum dinitrate was not irritating to rabbit eyes following instillation of the test material (0.1 ml) into the right eye of 3 animals.
Executive summary:

In an in vivo eye irritation study carried out in accordance with OECD Test Guideline 405, and to GLP, a 0.1 ml aliquot of undiluted tetraammineplatinum dinitrate was instilled into the conjunctival sac of the right eye of three male New Zealand White rabbits. The left eye was untreated to serve as a control. Following instillation the eyelids were held closed for 1 second. The ocular response was assessed at 1, 24, 48 and 72 hours. Corneal opacity, effects on the iris and conjunctival redness, chemosis and discharge were scored according to the Draize numerical scale. Any clinical signs of toxicity were noted and body weights were also recorded.

 

No corneal or iris effects were observed throughout the study. The only effect to be noted was minimal transient conjunctival redness in one treated eye one hour after treatment. No signs of toxicity were noted during the study and all animals showed expected bodyweight gain.

 

Based on the results of this study, tetraammineplatinum dinitrate should not be classified for eye irritation under EU CLP criteria (EC 1272/2008).

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

No relevant human irritation/corrosion data were identified.

 

Tetraammineplatinum dinitrate was tested for skin irritation potential in an in vitro reconstructed human epidermis model (EpiSkin assay) conducted in accordance with OECD Test Guideline 439, and to GLP. EPISKIN is a three-dimensional human skin model comprising a reconstructed epidermis with a functional stratum corneum. Its use for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability. Cell viability determination is based on cellular mitochondrial dehydrogenase activity, measured by MTT reduction and conversion into a blue formazan salt that is quantitatively measured after extraction from tissues. The reduction of cell viability in treated tissues is compared to negative controls and expressed as a %. If the resulting mean relative viability (as adjusted for intrinsic colour) is less than or equal to 50% of the negative control, the test substance is considered to be irritant to skin. Following a 15-minute exposure to the test substance, the test system skin cell viability was calculated to be greater than 50% (the average was 95% and the range was 85-100%), and it was therefore considered to be non-irritating to skin (Kiss, 2012). Under the conditions of this assay, tetraammineplatinum dinitrate does not require classification for skin irritation according to EU CLP criteria (EC 1272/2008).

 

In an in vivo eye irritation study carried out in accordance with OECD Test Guideline 405, and to GLP, a 0.1 ml aliquot of undiluted tetraammineplatinum dinitrate was instilled into the conjunctival sac of the right eye of three male New Zealand White rabbits. The left eye was untreated to serve as a control. Following instillation the eyelids were held closed for 1 second. The ocular response was assessed at 1, 24, 48 and 72 hours. Corneal opacity, effects on the iris and conjunctival redness, chemosis and discharge were scored according to the Draize numerical scale. Any clinical signs of toxicity were noted and body weights were also recorded. No corneal or iris effects were observed throughout the study. The only effect to be noted was minimal transient conjunctival redness in one treated eye one hour after treatment. No signs of toxicity were noted during the study and all animals showed expected bodyweight gain (Pooles, 2012). Based on the results of this study, tetraammineplatinum dinitrate should not be classified for eye irritation under EU CLP criteria (EC 1272/2008).

 

In an in vitro bovine corneal opacity and permeability (BCOP) assay conducted in accordance with OECD Test Guideline 437 and to GLP, tetraammineplatinum dinitrate was applied to isolated bovine corneas for 10 minutes, followed by an incubation period of 120 minutes. The In Vitro Irritancy Score (IVIS) calculated from individual scores for induced opacity (decreased light transmission through the cornea) and permeability (passage of sodium fluorescein dye through the cornea) was 1.7. A score of >55 is indicative of an ocular corrosive or severe irritant indicating that this substance is unlikely to have corrosive or severe irritant potential and would not be classified in Category 1 according to CLP criteria (Warren, 2012). The IVIS value falls below the cut-off value of 3 (no category) and hence no classification for eye irritation is required under EU CLP (EC 1272/2008). [The study was conducted subsequent to the revision to this test guideline, which incorporated this lower cut-off value of 3 (no category), hence the in vivo study was deemed necessary at the time to confirm the classification].

 

No respiratory tract irritation data were identified. A new study was not conducted as it is not a REACH Standard Information Requirement.

Justification for classification or non-classification

Based on the results of the available reliable in vitro skin irritation study, there is no requirement to classify tetraammineplatinum dinitrate for skin irritation under EC Regulation 1272/2008.

 

Based on the results of the available and reliable in vivo eye irritation study, supported by the in vitro investigation, tetraammineplatinum dinitrate does not require classification for eye irritation according to EU CLP criteria (EC 1272/2008).