Registration Dossier

Administrative data

Description of key information

Acute Oral Toxicity: DL50 >2000 mg/kg. GHS criteria: Cat. 5, CLP criteria not met.

Acute Dermal Toxicity: DL50 >2000 mg/kg. GHS criteria not met.

Acute Inhalation Toxicity: LC50 = 1.3 mg/L air (aerosol). Substance classifies as Acute tox. inhalation. Cat. 4.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04.Apr.07 - 30.Jul.07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
OECD Guideline for the Testing of Chemicals, Guideline 420 Acute Oral Toxicity, Fixed Dose Procedure, updated 17 December 2001
Deviations:
yes
Remarks:
The animals received A04C diet instead of TEKLAD 2014 as was indicated in the Study Plan. This had no ímpact on the Study.
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Version / remarks:
Commission Directiva 2004/73/EC of 29 April 2004. Annex 28. Method 8.1 bis
Qualifier:
according to guideline
Guideline:
other: Commission Directiva 2001/59/EC of 6 August 2001, Annex VI
GLP compliance:
yes
Test type:
acute toxic class method
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (range of 20 ± 5 ce) and in the dark.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The necessary amount of test item was weighed in a volumetric flask and distilled water was added gradually until the desired volume was reached.
The formulation was later transferred to an adequate container and kept in agitation. The necessary volume of vehicle was added to the formulation prepared on 3 May 2007 in the final container to make up the required volume. The vehicle added was first poured into the volumetric flask before being added to the final container.
Species:
rat
Strain:
Wistar
Remarks:
Wistar Hannover HsdRccHan: WIST rat
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Ibérica, S. L. {Ctra. Sant Miquel del Fai, km 3, 08182- Sant Feliu de Codines, Barcelona, Spain)
- Number of animals: 6 females
- Age when treated: 8-9 weeks
- Weight at study initiation: 158-164 g

CONDITIONS
Standard Laboratory Condltions.: Air-conditioned with target ranges for room temperature at 19-26 °C, relative humidity of 40-70% and 10-20 air changes per hour. Room temperature and relative humidity were monitored continuously. Values outside these ranges
occasionally occurred. These transient variations are not considered to have any influence on the study and therefore these data are not reported but are retained at RCC CIDA S.A. There was a 12-hour fluorescent light/12-hour dark cycle.

ACCOMMODATION
Sighting study: Individual in Makrolon type-5 cages {59.0 x 38.5 x 20.0 cm) with sawdust bedding {supplied by Harlan lnterfauna Ibérica, S.L.).
Main study: Groups of four or five in Makrolon type-5 cages {59.0 x 38.5 x 20.0 cm) with Lignocel 3-4 sawdust bedding (supplied by Harlan lnterfauna Ibérica, S.L.).

- Diet: Pelleted standard SAFE A04C rat/mouse maintenance diet (Scientific Animal Food & Engineering, Route de St. Bris, 89290-Augy, France, (Batch 70206}, Expiry date: 6 February 2008) ad libitum. Results of analyses for contaminants are archived at RCC CIDA S.A.
- Water: Tap water in bottles ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC CIDA S.A.
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Distilled water (Aqua B. Braun, Ref. 387875, Batches:7083B06, 7154B07
Doses:
lnitially, one rat was treated at the dose of 300 mg/kg. 8ecause no signs of toxicity were recorded in thís animal, another animal was treated at the dose of 2000 mg/kg
No. of animals per sex per dose:
One group of five females Wistar Hannover HsdRccHan rats (including one used in the initial sighting study)
Control animals:
not specified
Statistics:
No statistical analysis was used.
Preliminary study:
The animal treated at 2000 mg/kg is reported under "9.2 MAl N STUDY'.
No mortality nor clinical signs were observed in the animal treated at 300 mg/kg during the course of the study.
Key result
Sex:
female
Dose descriptor:
LD50
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
One animal died at the dosage level of 2000 mg/kg approximately one hour after administration.
Clinical signs:
The animal that died after administration showed decreased motor activity, abnormal gait, hunched back, decreased muscle tone, prostration, clonic convulsions, dyspnea and diarrhea during approximately 30 minutes following administration and before dying.
Hunched back and abnormal gait were recorded in other three animals during the 30 minutes after administration; up to 5 hours after administration in one animal, and during the observation period between 1 and 5 hours in the other two.
Hunched back and abnormal gait were recorded until day 4 after administration in one animal. Although in one of the animals abnormal gait persisted until day 9 and hunched back until day 11. Pallor was also observed until day 6, as well as red urine, dark feces and yellowish skin and mucosae until day 2-3 after administration in these two animals. Another animal showed abnormal gait and dark teces one day after treatment and pallor on days 3 and 4 of the observation period.
No clínica! signs were recorded from day 12 onwards.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
Right renal pelvis slightly dilated was recorded in one animal. No macroscopic alterations were recorded at the necropsies performed in the others animals that survived the treatment.
Hemorrhagic area in cranial surface of 0.5 x 0.5 cm diameter approximately and presence of product in the stomach were observed in the animal that died after the administration.
Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
DL50> 2000 mg/kg. Substance is not classified according CLP criteria.
Executive summary:

In the main study, one group of five females Wistar Hannover HsdRccHan rats (including one used in the initial sighting study) were treated with the test item by oral gavage administration at a dosage of 2000mg/kg body weight. The test item was diluted in vehicle (distilled water) at a concentration of 0,2g/mL and administered at a dosing volume of 10mL/ kg.

During the sighting study, one additional animal was dosed at 300mg/ kg without showing any signs of toxicity.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 -15. Mortality/viability was recorded at approximately 30minutes, 1, 2,3 and 5 hours after weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

The death of one of the animals treated at the dose of 2000mg/ kg, was recorded approximately one hour after the administration.

Among the animals treated at this dose hunched back and abnormal gait were recorded on the day of adminsitration.

The naimls thata died after the adminsitration also had decreased motor activity, abnormal gait, hunched back, decreased muscle tone, prostration, clonic convulsions, dyspnea and diarrhea.

Two of the animals thata survived the treatment showed huncled back, abnormal gait, pallor, dark red urine, dark feces andyellowish skin and mucus in the days following tretament. In one of the animals, these clinical signs were recorded at the most until day 6 after adminsitration. The other animal showed abnormal gait until day 9, and the hunched back until day 11 after adminsitration. Another animal showed abnormal gait and dark feces one day after administration, and pallor on days 3 and 4 after adminsitration.

No clinical signs were recorded from day 12 onwards.

Hemorrhagic area in cranial surface of 0.5 x 0.5 cm diameter and presence of product in stomach, were observed in the animal that died after administration.

In one animal was recorded right renal slightly diated. No macroscopical alterations were recorded at the necropsies performed in the others animals that survived the treatment.

The body weight of the animals was within the range commoly recorded for this strain and age.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19.Nov.2008 - 14.Jul.2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
OECD Guidelines for Testing of Chemicals, Section 4, Number 403: “Acute Inhalation Toxicity”, adopted May 12, 1981.
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
European Communities, Directive 92/69/EEC, Part B.2 “Acute Toxicity (Inhalation)”, published December 29, 1992 and European Communities Directive 93/21/EEC, April 27,1993 amending the aforementioned Directive
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
U.S. Environmental Protection Agency, Health Effects Test Guidelines OPPTS 870.1300, Acute Inhalation Toxicity, August 1998.
GLP compliance:
yes (incl. QA statement)
Species:
rat
Strain:
other: HanRcc:WIST(SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd. Laboratory Animal Services Wölferstrasse 4 4414 Füllinsdorf / Switzerland
- Number of Animals per Group: 5 males and 5 females
- Number of Groups: 2
- Females (if applicable) nulliparous and non-pregnant: [yes/no]
- Age at study initiation: Group 1 and 2: Males 10 weeks, Females 11 weeks
- Weight at study initiation:
Group 1: Males: 251.7 to 267.3 g, Females: 210.2 to 228.7 g
Group 2: Males: 257.3 to 287.0 g, Females: 205.9 to 213.1 g
The weight variation did not exceed ± 7.5% of the mean weight of the corresponding sex.
- Identification: By unique cage numbers and individual unique tail numbers written with an indeligible felt-tip pen.
- Diet (e.g. ad libitum): Animals had ad libitum access to a pelleted standard Kliba-Nafag 3433 rat maintenance diet (Provimi Kliba AG, 4303 Kaiseraugst, Switzerland) batch nos. 76/08 and 31/08 except during the period when they were restrained in exposure tubes. Results of the analyses for contaminants and their limits of acceptability are archived at Harlan Laboratories Ltd.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum in water bottles, except during the period when they were restrained in exposure tubes. Results of
representative analyses for contaminants are archived at Harlan Laboratories Ltd.
- Acclimation period: Performed under Harlan Laboratories Study B68308 for at least five days under laboratory conditions, after clinical health examination. Only animals without any visible signs of illness were used for the study. A further observation of clinical signs was performed on each day of exposure, before exposure start.

ENVIRONMENTAL CONDITIONS
- Conditions: Standard laboratory conditions. Air-conditioned with 10 - 15 air changes per hour, continuously monitored environment with target temperature of 22 ± 3 °C, a target relative humidity of 30 - 70% and a 12 hour fluorescent light / 12 hour dark cycle. Values for humidity and temperature out of the target range occasionally occurred, usually following room cleaning, and are considered not to have any influence on the study. Therefore these data are not reported but retained at Harlan Laboratories Ltd. A radio program was played during the most of the light period.
- Accommodation: Animals were housed in groups of 5 of the same sex in Makrolon® type-IV cages with wire mesh tops and standard softwood bedding ("Lignocel", Schill AG, 4132 Muttenz, Switzerland).
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Remarks:
flow-past exposure
Vehicle:
not specified
Mass median aerodynamic diameter (MMAD):
>= 2.27 - <= 2.65 µm
Geometric standard deviation (GSD):
>= 1.98 - <= 2.41
Remark on MMAD/GSD:
Particle Size Distribution
The Mass Median Aerodynamic Diameters (MMAD) obtained from two gravimetric measurements of particle size distribution during each exposure were similar (Group 1: MMAD = 2.27 and 2.84 μm, GSD = 1.98 and 2.41; Group 2: 2.37 and 2.65 μm, GSD = 2.23 and 2.30). This led to the conclusion that the particle size of the generated aerosol was fairly stable during the whole exposure periods. The MMADs for both groups were within the target range of 1 to 4 μm, thus deposition of the particles can be assumed to have occurred in both the upper and the lower respiratory tract. Hence, the particle size distribution and MMADs obtained were considered to be appropriate for acute inhalation toxicity testing.
Data on particle size distribution are presented in the tables in page 24 from the study report.
Details on inhalation exposure:
TREATMENT
- Room: Inhalation laboratory no. 222, Harlan Laboratories Ltd., Füllinsdorf
- Method: Inhalation by nose-only, flow-past exposure
- Rationale for Method: Inhalation is a possible route of human exposure.
- Frequency of Administration: Single, 4-hour exposure period. Exposure of Group 1 was interrupted twice for a total of 5 minutes for changing of the nebulizer or a connection.
Nevertheless, the animals were exposed for a period of 4 hours as those interruptions were accounted for.
- Rationale for Aerosol Concentration:
--> Group 1: The target concentration of approximately 5 mg/L air (actual concentration of respirable substances) is the recommended concentration (OECD 403, “Acute Inhalation Toxicity”).
--> Group 2:
The target concentration of 1 mg/L air (actual concentration of respirable substances) for 4 hours is the limit concentration for Category 4 (GHS ST/SG/AC.10/30 2003).
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Remarks on duration:
Single, 4-hour exposure period. Exposure of Group 1 was interrupted twice for a total of 5 minutes for changing of the nebulizer or a connection. Nevertheless, the animals were exposed for a period of 4 hours as those interruptions were accounted for.
Concentrations:
The chemical aerosol concentrations determined were 6.1 mg/L air (Group 1) and 1.3 mg/L air (Group 2) as targeted.
No. of animals per sex per dose:
Group 1
Males 1-5 slightly above 5 (mg/L air)
Females 6-10 slightly above 5 (mg/L air)

Group 2
Males 11-15 slightly above 1 (mg/L air)
Females 16-20 slightly above 1 (mg/L air)
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
After 14 days: 04-Dec-2008 (Group 1)
22-Jan-2009 (Group 2)
All surviving animals were transferred to the pathology unit and anaesthetised by an intraperitoneal injection of sodium pentobarbitone and killed by exsanguination. All animals were necropsied and abnormalities recorded. The animals that died spontaneously during the exposure were transferred to the pathology unit and necropsied as soon as possible.
Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered 4% formaldehyde solution. All collected organs/tissues are available for histopathological examination, if requested by the Sponsor under separate contractual agreement.
Tissue/Organs colleted: Head with nasopharyngeal tissues, Larynx, Lungs, instilled via trachea with formalin at approximately 30 cm H2O pressure, Trachea and All gross lesions

OBSERVATIONS
The day of exposure was named test day 1 and counting continued for the subsequent days of observation.
--> Viability /Mortality
Observations for viability were recorded once before exposure on the day of exposure, once per hour during exposure, once after exposure on test day 1 and twice daily during the observation period.
--> Clinical Signs
Each animal was examined once per hour during exposure, once after exposure on test day 1 and once daily during the observation period. Observations were detailed and carefully recorded using explicitly defined scales as appropriate. Only grossly abnormal signs were detectable during exposure, as the animals were restrained in the exposure tubes.
--> Body Weights
The body weight of each animal was recorded on test days 1 (before exposure), 4, 8 and 15 (before necropsy).
Statistics:
No statistical analysis was performed as only two groups were allocated to the study.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
>= 1.3 - <= 6.1 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
Observations for viability were recorded once before exposure on the day of exposure, once per hour during exposure, once after exposure on test day 1 and twice daily during the observation period.
Three males died during the exposure at 6.1 mg/L air and one female within two hours after the end of exposure period. Two females treated at 6.1 mg/L air were found dead on day 2 of the observation period. One female was found dead on day 2 after exposure at 1.3 mg/L air. All other animals survived the scheduled observation period
Clinical signs:
other: Each animal was examined once per hour during exposure, once after exposure on test day 1 and once daily during the observation period. Observations were detailed and carefully recorded using explicitly defined scales as appropriate. Only grossly abnormal
Body weight:
The body weight of each animal was recorded on test days 1 (before exposure), 4, 8 and 15 (before necropsy).
Marked body weight loss was noted in all surviving animals treated at 6.1 mg/L air between test days 1 and 4. Thereafter normal body weight gain was recorded in these animals. Marginal body weight loss was seen in two males and one female treated at 1.3 mg/L air from test day 1 to 4. Thereafter these animals showed a normal body weight development. No effects on body weight were recorded in the remaining animals.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
LC50 = 1.3 mg/L air (aerosol). Substance classifies as Acute tox. inhalation. Cat. 4.

Executive summary:

Two groups of five male and five female albino rats [HanRcc:WIST(SPF)] each were exposed by nose-only, flow-past inhalation to the test item at a chemically determined mean concentration of 6.1 mg/L air or 1.3 mg/L air, respectively. All animals were observed for clinical signs and mortality during the inhalation exposure and the observation period of 14 days. Body weights were recorded prior to exposure on test day 1, and during the observation period on test days 4, 8 and 15 before necropsy. On day 15 all surviving animals were sacrificed and necropsied.

The ranges of aerosol concentration, temperature, relative humidity, oxygen content and airflow rate measured during the exposure were considered to be satisfactory for a study of this type. In addition, the test item was considered to be respirable to rats.

Three males and one female died during or shortly after the exposure at 6.1 mg/L air. Two females treated at 6.1 mg/L air and one female treated at 1.3 mg/L air were found dead on day 2 of the observation period. All other animals survived the scheduled observation period.

Mortality:

 Aerosol Concentration  Males  Females Both sexes 
 1,3mg/L air 0%   20% 10%  
  6,1 mg/L air 60%   60%  60%

Salivation, decreased spontaneous activity, rales and ruffled fur were observed in most of the animals of both groups. Tachypnea was seen in animals exposed at 6,1mg/ L air.

Marked body weight loss was observed in all surviving animals treated at 6.1 mg/L air between test days 1 and 4. Thereafter these animals showed a normal body weight development.

Marginal body weight loss was recorded in two males and one female treated at 1.3 mg/L air between test days 1 and 4. Thereafter these animals showed a normal body weight development.

There were no macroscopic findings that were considered to be related to treatment with the test item.

In conclusion, the LC50 of Servalesa L60 obtained in this study was estimated to be between 1.3 mg/L air and 6.1 mg/L air (chemically determined mean aerosol concentration). There was no indication of relevant sex-related differences in toxicity of the test item.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Value:
1 300 mg/m³

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04.Apr.2007 - 30.Aug.2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
- During the study, the temperature in the animals house was 19-28 °C and the relative humidity 35-70%. - The animals received A04C diet instead of TEKLAD 2014 as indicated in the Study Plan. These deviations had no impact on the Study
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
Commission Directive 92/69/EEC of 31 July 1992, Annex, Part B, Method 8.3, published in the Official Journal on 29 December 1992
Qualifier:
according to guideline
Guideline:
other: Directive 2003/63/EC
Version / remarks:
Commission Directive 2003/63/EC of 25 June 2003 amending Directive 2001/83/EC of the European Parliament and of the Council on the Community code relating to medicinal products for human use.
GLP compliance:
yes
Test type:
standard acute method
Specific details on test material used for the study:
TEST ITEM PREPARATION
Dose levels were in terms of the test item as supplied by the Sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Approximately 24 hours before adminsitration of the test item, the back of each animal was shaved using an electric razor, from the scapula area to the lumbar region.
The test item was applied on an area representing around 10% of the total body surface area.
The test item was placed on square of hydrophilic gauze of approximately 6 x 5 cm and applied to the corresponding test area. The gauze was then covered with a hypoallergenic microporous ahesive band. This route was chosen to study the possible toxic effects produced by skin exposition to the test item.
Finally a strip, of gauze was wrapped around the trunk of the rat in order to hold the patches in place.
The band was fixed to the body of the animal using adhesive tape.
The aim of this semioclusive bandage was to allow good skin absorption of the test item and keep animals from ingesting it.
The bandage and the patches were removed 24 hours after expousure. The remains of the product were removed with distilled water.
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Interfauna Ibérica, S.L. (Ctra. Sant Miquel del Fai, km 3, 08182-Sant Feliu de Codines, Barcelona, Spain
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 7-8 weeks
- Weight at study initiation: 202-227g
- Number of animals. 14
- Identification: By means af an ear-punch technique
- Veterinaty inspection: During acclimatization, the animals were examined by a veterinary surgeon. Only animals without any visible signs of illness were used for the treatment.
- Acclimation period: From 7 to 8 days
- Housing: in groups of maximum five rats of the same sex and treatment group, in Makrolon type-5 (59.0x 38.5 x 20.0cm) cages with Lignocel 3-4 sawdust bedding (supplied by Harlan Interfauna Ibérica, S.L.). At the time of administration, the animals will be moved to individual cages with grilled floor to avoid contact with sawdust or other alíen materials that could lead to skin irritation. Once it is checked there is no skin irritation, the grilled floor will be replaced by sawdust again.
- Fasting period before study: not specified
- Diet (e.g. ad libitum): Pelleted standard SAFE A04C rat/mouse maintenance diet (Scientific Animal Food & Engineering, Route de St.Bris, 89290-Augy, France, Batches no.60928 and 70206, Expiry dates: 28 September 2007 and 6 February 2008, respectively) ad libitum. Results of analyses for contaminants are archived at RCC CIDA S.A.
- Water (e.g. ad libitum):Tap water in bottles adbilitum. Results of bacteriological chemical and contaminant analyses are archived at RCC CIDA S.A.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 (monitored)
- Humidity (%): 35-70 (occasionally reaching 80%) (monitored)
- Air changes (per hr): 10-20
- Photoperiod (hrs dark / hrs light): 12-hour fluorescent light/12-hour dark cycle
Type of coverage:
semiocclusive
Vehicle:
other: square of hydrophilic gauze
Details on dermal exposure:
TREATMENT
Approximately 24 hours before administration of the test item, the back of each animal was
shaved using an electric razor, from the scapula area to the lumbar region.
The test item was applied on an area representing around 1 0% of the total body surface area.
The test item was placed on a square of hydrophilic gauze of approximately 6 x 5 cm and applied to the corresponding test area. The gauze was then covered with a hypoallergenic microporous adhesive band. This route was chosen to study the possible toxic effects produced by skin exposition to the test item.
Finally, a strip of gauze was wrapped around the trunk of the rat in order to hold the patches in
place.
The band was fixed to the body of the animal using adhesive tape.
The aim of this semiocclusive bandage was to allow good skin absorption of the test item and
keep animals from ingesting it.
The bandage and the patches were removed 24 hours after exposure. The remains of the product were removed with distilled water.

DOSE LEVELS
In the sighting study, the dose of 2000 mg/kg was administered.
Since no mortality was observed, this dose was administered in the main study.
Duration of exposure:
24 hours: The bandage and the patches were removed 24 hours after exposure. The remains of the product were removed with distilled water.
Doses:
In the sighting study, the dose of 2000 mg/kg was administered. Since no mortality was observed, this dose was administered in the main study.
No. of animals per sex per dose:
A single dose of test ítem was administered to five males and five females
Control animals:
not specified
Details on study design:
SIGHTING STUDY
The test ítem was administered by the dermal route on a single occasion, to two males and two females.
The rats were observed after the treatment to record the clínica! responses and mortality. They were sacrificed 14 days after the treatment, when sufficient information was obtained. No autopsies were performed nor any tissues were kept.

MAIN STUDY
A single dose of test ítem was administered to five males and five females.

OBSERVATIONS
Mortality/Viability: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (in common with the clínica! signs) and twice daily during days 2-15.
Body weights: On test days 1 (prior to administration), 8 and 15.
Clínica! signs: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test da y 1. Once daily d uring days 2-15.
Statistics:
No statistical analysis was used.
Preliminary study:
No mortality was recorded.
No clinical signs were recorded.
No dermal alterations were recorded on removing the semiocclusive patches.
The body weight of the animals was within the range commonly recorded for this strain and age.
Key result
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No deaths ocurred at the dose level of 2000 mg/kg during the study.
Clinical signs:
No clinical signs were observed during the course of the study.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necrospy
Other findings:
All surviving animals were killed at the end of the observation period by an intraperitoneal injection of sodium pentobarbital at the dese of 200 mg/kg body weight and at volume of 10 mL/kg. A necropsy was carried out on all the animals of the main study. This included the examination of the intact animal and all superficial tissues, followed by the in situ observation of the cranial, thoracic and abdominal cavities.
Interpretation of results:
GHS criteria not met
Conclusions:
Since no mortality was recorded after administration of the test item at the dose of 2000mg/kg, the LD50 of the test item has been found to be higher than the above-mentioned dose when administered by dermal route to rats, and therefore it is not necessary to assign it a risk phrase.
Executive summary:

In the main study, one group of ten Hsd: Sprague Dawley "SD" rats were treated with the test item by dermal route at a dosage 2000mg/kg body weight.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at appoximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 -15. Mortality/viability was recorded and approximately 30minutes, 1,2, 3 and 5 hours after administration on test day 1 (with the clinical signs)and twice daily during days 2 -15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period. No clinical signs were observed during the course of the study.

No dermal alterations were recorded on removal of the semiocclusive patches after 24 hours of exposure.

The body weight of the animals was within the range conmonly recorded for this strain and age.

No macroscopic findings were recorded at necropsy.

Additional information

Although the physical state described on section 1.2 (composition) and 1.4 (Appearance/ physical state/ colour) for the substance Reaction mass of copper digluconate, sodium sulphate and copper sulphate is solid, the manufacturing method (described in section 3.5.1.) only allows substance synthetisation as a dissolution, containing at most a 50% of Reaction mass of copper digluconate, sodium sulphate and copper sulphate.

Therefore, studies for physical hazards determination, were developed using solid substance form, whereas for toxicology and ecotoxicology hazards, the dissolution (≤ 50% Reaction mass of copper digluconate, sodium sulphate and copper sulphate) form was used as stated on guidelines.

In consequence, the substance has been classified taking into account that it always will be in a dissolution in a maximum of 50% of concentration.

Justification for classification or non-classification

The oral LD50 was above 2000 mg/kg bw. Therefore, the substance does not classifiy.

The dermal LD50 was above 2000 mg/kg bw. Therefore, the substance does not classifiy.

The inhalation between 1.0 and 5.0 mg/kg bw (LC50=1.3 mg/L air (aerosol). Therefore, the substance is classified as Acute Tox. Cat. 4 (Inhalation)