Tetraammineplatinum dinitrate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

22 September 2004 to 18 October 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Conducted according to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

yes

Remarks:

deviations in levels of relative humidity, deviations in minimum temperature and deviations in mouse strain [OECD recommends CBA/Ca or CBA/J mice] were not thought to affect study integrity

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Platinum(2+) tetraammine (SP-4-1) diacetate

IUPAC Name:

Platinum(2+) tetraammine (SP-4-1) diacetate

Constituent 2

Reference substance name:

127733-97-5

Cas Number:

127733-97-5

IUPAC Name:

127733-97-5

Details on test material:

- Name of test material (as cited in study report): platinum(2+) tetraammine (SP-4-1) diacetate
- Substance type: no data
- Physical state: white crystalline powder
- Analytical purity: 95.2%
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: Pt 48.75%, Cl 0.029%
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: GB 379
- Expiration date of the lot/batch: 28 June 2005
- Stability under test conditions: not indicated
- Storage condition of test material: at room temperature in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River France, l'Arbresle Cedex, France
- Age at study initiation: approximately 11 weeks
- Weight at study initiation: within 20% of the sex mean
- Housing: individual
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.4 to 21.7 [OECD recommends a minimum temperature of 19 °C]
- Humidity (%): 29 to 94 [OECD specifies 30 to 70%, ideally 50 to 60%]
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: no data

Study design: in vivo (LLNA)

Vehicle:

propylene glycol

Concentration:

Concentrations of 25 and 50% were used in the preliminary irritation study. Concentrations of 5, 25 and 50% were used in the subsequent local lymph node assay.

No. of animals per dose:

Preliminary irritation study: 1
Local lymph node assay: 5

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: no data
- Irritation: two mice were treated with a dermal dose of platinum(2+) tetraammine (SP-4-1) diacetate at either 25 or 50% on each ear over three consecutive days. Ears were examined four hours after the last exposure and graded.
- Lymph node proliferation response: no data

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: local lymph node assay
- Criteria used to consider a positive response: a substance may be regarded as a skin sensitiser if the results indicate a stimulation index of at least 3

TREATMENT PREPARATION AND ADMINISTRATION:
INDUCTION (days 1, 2 and 3)
25 µl of the test substance was applied to the dorsum surface of both ears at 5%, 25% or 50% at approximately the same time each day. Controls recieved vehicle alone.

TREATMENT (day 6)
Animals were injected via the tail vein with 0.25 ml sterile phosphate buffered saline (PBS) containing 20 µCi of 3H-methyl thymidine. Approximately five hours later, animals were sacrificed by an overdose of pentobarbital via intraperitoneal injection. A gross macroscopic examination was perfomed. The draining lymph node of each ear was excised, nodes were examined visually and any abnormalities were recorded.
For each animal, a single cell suspension of lymph node cells was prepared. DNA was precipitated and processed for radioactivity.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

no data

Results and discussion

Positive control results:

Historical lab data performed in July/August 2004 involved the treatment of groups of female mice treated with alpha-hexylcinnamicaldehyde (tech. 85%) at concentrations of 5, 10 or 25%, using a similar procedure. SI values of 1.0, 3.2 and 7.1 were recorded, and an EC3 value of 9.5% was calculated.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: Group 1 - propylene glycol (vehicle control): SI +/- SD = 1.0 Group 2 - 5% test substance: SI +/- SD = 1.4 Group 3 - 25% test substance: SI +/- SD = 1.3 Group 4 - 50% test substance: SI +/- SD = 1.4

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see Remark

Remarks:

Group 1 - propylene glycol (vehicle control): 700, 197, 151, 337 and 571 (five animals). Median DPM/animal value = 337. Group 2 - 5% test substance: 381, 362, 525, 482, 618 (five animals). Median DPM/animal value = 482. Group 3 - 25% test substance: 645, 285, 466, 377 (four animals - an anomalous value of 4766 was recorded for a further mouse, but rejected based on abnormalities in the right eye and an extremely large right lymph node). Median DPM/animal value = 422. Group 4 - 50% test substance: 805, 1555, 4020, 738, 416. Median DPM/animal value = 805.

Any other information on results incl. tables

Skin irritation reactions were graded 0 (corresponding to "no erythema" and "no oedema") for all tested animals at all doses.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In a guideline LLNA, to GLP, platinum(2+) tetraammine (SP-4-1) diacetate was non-sensitising at concentrations of up to 50%.

Executive summary:

In a guideline local lymph node assay, conducted to GLP, 25µl platinum(2+) tetraammine (SP-4-1) diacetate in propylene glycol was applied once daily to the skin of both ears of female CBA mice (5/group), at 0, 5, 25 or 50%, for three consecutive days. Mice were monitored for mortality and overt signs of toxicity, as well as changes in body weight. Skin irritation was assessed and graded three to four hours after the final dose (on day three). Three days after this induction period (on day six), mice were given tail vein injections of radiolabelled thymidine, and were sacrificed after approximately five hours. Lymph nodes were excised and processed for radioactivity.

 

No skin irritation was seen as a result of treatment with platinum(2+) tetraammine (SP-4-1) diacetate. Lymph nodes were a normal size, with the exception of one animal given the test substance at 25% and showing eye abnormalities. The anomalous results for this animal were subsequently discounted. No mortality or substance-related toxicity was observed over the course of the experiment.

 

Median disintegrations per minute/animal values were 337, 482, 422 and 805 for groups of mice given platinum(2+) tetraammine at concentrations of 0, 5, 25 and 50% respectively. SI values were 1.4, 1.3 and 2.4 for animals given respective concentrations of 5%, 25% and 50%. An SI value of over three was judged to be indicative of a substance being sensitising. Hence, under the conditions of this assay, platinum(2+) tetraammine (SP-4-1) diacetate was considered non-sensitising.

 

Based on the results of this study, no classification for skin sensitisation is required according to EU CLP criteria (EC 1272/2008).