Iron(1+), chloro[dimethyl 9,9-dihydroxy-3-methyl-2,4-di(2-pyridinyl-kN)-7-[(2-pyridinyl-kN)methyl]-3,7-diazabicyclo[3.3.1]nonane-1,5-dicarboxylate-kN3,kN7]-, chloride

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 8 December 2003 to 07 June 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

The test item was incorrectly and therefore a dose of 800 mg/kg was dosed instead of 2000 mg/kg, as stated in the study plan. it was decided that the study procedure should continue with a 2000 mg/kg dose to enable an LD50 value to be determined.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,5-dimethyl 3-methyl-9-oxo-2,4-bis(pyridin-2-yl)-7-[(pyridin-2-yl)methyl]-3,7-diazabicyclo[3.3.1]nonane-1,5-dicarboxylate dichloroiron hydrate

EC Number:

610-388-9

Cas Number:

478945-46-9

Molecular formula:

C28H31N5O6FeCl2

IUPAC Name:

1,5-dimethyl 3-methyl-9-oxo-2,4-bis(pyridin-2-yl)-7-[(pyridin-2-yl)methyl]-3,7-diazabicyclo[3.3.1]nonane-1,5-dicarboxylate dichloroiron hydrate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: SEAC sample number S2539801
- Expiration date of the lot/batch: 1 January 2005
- Purity test date: The supporting data for purity of the test item was not made available at the time of issuing this report. However, sponsor has adressed this in a GLP compliant study, SEAC Study Reference Number AC030449

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature (range of 20±3 deg celsius), protected from light
- Stability under test conditions:stability of the dosing solution was adressed by ensuring fresh solutions were made immediately prior to dosing.
- Solubility and stability of the test substance in the solvent/vehicle: A solubility trial was carried out to determine the choice of vehicle.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, laboratory Animal Services
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 weeks (male) ; 12 weeks (female)
- Weight at study initiation: male : 245.8 to 279.4g female : 229.7g to 252.6g
- Fasting period before study: not specified
- Housing:In groups of 4 or 5 in Makrolon type-4 cages with wire mesh tops and standard softwood bedding during acclimation period. Individually in Makrolon type 3 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch nos. 54/03 ad libitum
- Water (e.g. ad libitum): community tap water ad libitum
- Acclimation period: 5-6 days under laboratory conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3 deg celsius
- Humidity (%): 30-70% relative humidity
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark, music duting the daytime light period

IN-LIFE DATES From: : 9 December 2003 To: 30 March 2004

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

purified water

Details on dermal exposure:

TEST SITE
- Area of exposure: back of the animals
- % coverage: approximately 10% of the total body surface
- Type of wrap if used: elastic adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The skin was flushed with lukewarm tap water and dried with disposal paper towels.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4 mL/kg bodyweight
- Concentration (if solution): 0.2 g/mL and 0.5 mg/mL
- Constant volume or concentration used: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 4mL/kg bw
- Concentration (if solution): not specified
- Lot/batch no. (if required): not specified
- Purity: not specified

Duration of exposure:

24 hours

Doses:

800 mg/kg and 2000 mg/kg

No. of animals per sex per dose:

5 animals were used per sex and per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality : daily during the acclimatization period except in the five 2000 mg/kg treated males, twice daily during days 1-15
Body weight : at day 1, 8 and 15
Clinical signs : daily during acclimatization, and at 1,2,3 and 5 hours after administration the day 1
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, macroscopic examination

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured

Clinical signs:

other: Dose of 800 mg/kg bw : Slight scaling was observed in one female at the 9- and 10-day reading and in one female between day 7 and the end of the study period (day 15). Two females were seen with focal erythema on the test site from the 7-day reading up to

Gross pathology:

The caecum was distended with gas in one 800 mg/kg treated male at scheduled necropsy. There were no macroscopic in any of the other animals at scheduled necropsy.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to the results of the study, the registered substance stainless E-700-2003 did not induced toxicity nor mortality in acute dermal toxicity test. Hence, the LD50 was defined as greater than 2000 mg/kg bw (the high dose level used).

Executive summary:

This GLP-compliant study was performed in order to assess the potential acute toxicity by dermal contact in rats (according to the 402 OECD guideline method).

The test item was dermally tested at 800 and 2000 mg/kg by using 5 males and 5 females HanBrl: WIST (SPF) rats per dose. The test item was diluted in vehicle (purified water) at a concentration of 0.2 and 0.5 mg/ml , respectively administered at a volume dosage of 4 mL/kg. The application period was 24 hours.

The animals were examined daily for mortality, clinical signs and body weight was measured too (day 1, 8 and 15). Necropsy and macroscopic examination were performed at the day 15.

Dose of 800 mg/kg bw : Slight scaling was observed in one female at the 9- and 10-day reading and in one female between day 7 and the end of the study period (day 15). Two females were seen with focal erythema on the test site from the 7-day reading up to the end of the study period. No clinical signs were noted in the females. No clinical signs and no local signs were recorded in the males during the course of the study. Dose of 2000 mg/kg bw : Slight general erythema was observed in four of the females on test day 2 after removal of the dressing. One female still showed a slight erythema on test day 3. Slight focal erythema was seen in two females from test day 7 to the end of the study.No clinical signs were noted in the females. No clinical signs and no local signs were recorded in any of the males during the course of the study, however, a slight yellow discolouration was observed on the skin of all males on day 2 which persisted in four of the animals for at least 3 days.

Loss of bodyweight (0.6% to 4.9%) was observed in one animal of the 800 mg/kg group (female) and in three 2000mg/kg females at the day 8-day reading. By the end of the study, the animal had regained some of weight that was lost. In spite of this body weight loss, the body weight of all animals during the course of the study was within the range commonly recorded for this strain and age

The caecum was distended with gas in one 800 mg/kg treated male at scheduled necropsy. They were no macroscopic in any of the other animals at scheduled necropsy.

According to the results of the study, the registered substance stainless E-700-2003 did not induced toxicity nor mortality in acute dermal toxicity test. Hence, the LD50 was defined as greater than 2000 mg/kg bw (the high dose level used).