Registration Dossier

Administrative data

Description of key information

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:
18 November - 28 December 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred on two days (39.67 and 38.65%). Evaluation: Laboratory historical data do not indicate an effect of the deviations.
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
May 2008, including the most recent amendments
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred on two days (39.67 and 38.65%). Evaluation: Laboratory historical data do not indicate an effect of the deviations.
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred on two days (39.67 and 38.65%). Evaluation: Laboratory historical data do not indicate an effect of the deviations.
Qualifier:
according to
Guideline:
other: JMAFF Notification No 8147
Version / remarks:
November 2000, including the most recent parital revisions
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred on two days (39.67 and 38.65%). Evaluation: Laboratory historical data do not indicate an effect of the deviations.
GLP compliance:
yes
Test type:
acute toxic class method
Species:
rat
Strain:
Wistar
Remarks:
Crl:WI (Han) (outbred, SPF-Quality)
Sex:
female
Details on test animals and environmental conditions:
Animal Husbandry
Conditions
Environmental controls for the animal room were set to maintain 18 to 24°C (actual range 20.1 – 20.8 °C), a relative humidity of 40 to 70% (actual range 39 – 48%), at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.
Accommodation
Group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
Acclimatization period was at least 5 days before start of treatment under laboratory conditions.
Diet
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
Water
Free access to tap water.
Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

Age and body weight
Young adult animals (approx. 8-9 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification
Earmark and tail mark
Health inspection
At least prior to dosing. It was ensured that the animals were healthy and without any abnormality that might have affected the study integrity.
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
The vehicle was selected based on trial preparations performed at Charles River Den Bosch and on test item data supplied by the Sponsor. There was no information available regarding the solubility or stability in vehicle.

The preparations (w/w) were kept at room temperature and were dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test item. The concentration of the test item in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight. In order to obtain homogeneity, the test item (preparations) were stirred for 15 minutes.

Oral gavage, using plastic feeding tubes. The test item preparations were stirred on a magnetic stirrer during dosing.
10 mL/kg b.w. for each dose.
Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test item. Water was available ad libitum.

Single dosage on Day 1.
Doses:
2000 mg/kg (10 mL/kg) body weight.
300 mg/kg (10 mL/kg) body weight.
50 mg/kg (10 mL/kg) body weight.
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Mortality/Viability
Twice daily. The time of death was recorded as precisely as possible.
Body weights
Days 1 (pre-administration), 8 and 15.
Clinical signs
At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 4: grading slight (1) to very severe (4) Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
Necropsy
Animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
Statistics:
/
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 50 - <= 300 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Remarks:
Based on OECD 423 test guideline
Effect level:
200 mg/kg bw
Based on:
test mat.
Mortality:
At 2000 and 300 mg/kg all animals were found dead on Day 1 post-treatment.
Clinical signs:
At 2000 mg/kg, no clinical signs were noted.
At 300 mg/kg, hunched posture was noted for all animals on Day 1.
At 50 mg/kg, hunched posture, uncoordinated movements and/or piloerection were noted for all animals on Day 1. The surviving animals had recovered from the symptoms by Day 2.
Body weight:
The mean body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. Laboratory historical data indicate weight ranges of 162-212, 161-213 and 183-234 grams for untreated animals of 9, 10 and 11 weeks old, respectively.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
The oral LD50 value of CH02906 in Wistar rats was established 50-300 mg/kg body weight.
According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 200 mg/kg body weight.

Based on these results:
• according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), CH02906 should be classified as: Toxic if swallowed (Category 3) for acute toxicity by the oral route.
• according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), CH02906 should be classified as Category 3 and should be labeled as H301: Toxic if swallowed.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
200 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:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Version / remarks:
September 2009
Deviations:
yes
Remarks:
During characterization of the test atmosphere, the weight of the used test item was not correctly noted in the raw data, therefore no nominal concentration and generation efficiency could be determined.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Before use the test item was grinded with an automatic grinder (ZM-100, Retsch, Ochten, The Netherlands).
Species:
rat
Strain:
Wistar
Remarks:
Crl: WI(Han)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: approximately 9-10 weeks old
- Weight at study initiation:
Males: 262 to 315 g.
Females: 178 to 223 g
- Fasting period before study: no
- Housing:
On arrival and following assignment to the study, animals were group housed (up to 5 animals of the same sex and same exposure group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room(s) in which the animals were kept were documented in the study records.
Animals were separated during designated procedures/activities. Each cage was clearly labeled.
- Diet (e.g. ad libitum):
Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility.
It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum):
Municipal tap-water was freely available to each animal via water bottles.
Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility.
It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period: The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.

ENVIRONMENTAL CONDITIONS
Target temperatures of 18 to 24°C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 19 to 20°C with an actual daily mean relative humidity of 50 to 65%. A 12‑hour light/12‑hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

IN-LIFE DATES: From: To:
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
>= 3.2 - <= 4.5 µm
Geometric standard deviation (GSD):
>= 1.8 - <= 2.2
Remark on MMAD/GSD:
The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined at least twice during the exposure period. At 1 mg/L, the MMAD was 3.2 µm (gsd 1.9) and 3.3 µm (gsd 2.2). At 3.9 mg/L, the MMAD was 4.0 µm (gsd 1.9), 4.5 µm (gsd 1.8) and 4.1 µm (gsd 2.0). Agglomeration of aerosol particles at the high concentration might have resulted in the MMAD values to fall outside the recommended range of 1 - 4 µm. The MMAD only slightly exceeded this range and there was no evidence for item deposition in the upper airways. Since good distribution throughout the lung requires particles with an aerodynamic diameter between 1 and 5 μm, it can be assumed that sufficient deposition in the lower respiratory tract occurred during the exposure.
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The design of the exposure chamber is based on the directed flow nose only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983).
- Exposure chamber volume: /
- Method of holding animals in test chamber: polycarbonate restraining tubes
- Source and rate of air: The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min.
- Method of conditioning air: /
- System of generating particulates/aerosols: An aerosol was generated by administering the test item to a stream of pressurized air using a combination of a spiral feeder (Randcastle Extrusion Systems, Cedar Grove, NJ, USA) and air mover (AIR-VAC, Milford, CT, USA). Outside air was pressurized and passed through several filters before use. The aerosol was passed through a series of two cyclones (1 mg/L) or an elutriator and three cyclones (3.9 mg/L), allowing larger particles to settle, before it entered the exposure chamber.
From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.
- Method of particle size determination: The particle size distribution was characterized at least twice during each exposure period. The samples were drawn with a flow of 2 L/min. from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (TE-290-GF. Tisch Environmental, Cleves, Ohio, USA) and a fiber glass back-up filter (SEC-290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test item collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined based on OECD guidance document No. 39.
- Treatment of exhaust air: From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.
- Temperature, humidity, pressure in air chamber: The temperature and relative humidity were measured with a humidity and temperature indicator (E+E Elektronik, Engerwitzdorf, Austria) and recorded after the animals were connected to the exposure chamber and at 30 minute intervals after initiation of exposure. The probe was inserted in a tube mounted in one of the free animal ports of the exposure chamber. The temperature of the atmosphere during the exposures was between 21.6 and 22.0oC. The relative humidity was between 27 and 36% which was considered appropriate for this relatively short 4 hours exposure duration.

TEST ATMOSPHERE
- Brief description of analytical method used: A total of 29 and 21 representative samples were taken for determination of the actual concentration during exposure at 1 mg/L and the technically maximum attainable concentration, respectively. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber. Samples were drawn through a glass fiber filter (type APFC04700, Millipore, Billerica, MA, USA). Sample volumes were measured by means of a dry gas meter (type G 1.6, Actaris Meterfabriek B.V., Dordrecht, The Netherlands). The collected amount of test item in the air sample was measured gravimetrically.
- Samples taken from breathing zone: yes

VEHICLE
- Composition of vehicle (if applicable): air
- Concentration of test material in vehicle (if applicable): 1 and 3.9 mg/L
- Justification of choice of vehicle: standard protocol
- Lot/batch no. (if required): /
- Purity: /

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: /
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): At 1 mg/L, the MMAD was 3.2 µm (gsd 1.9) and 3.3 µm (gsd 2.2). At 3.9 mg/L, the MMAD was 4.0 µm (gsd 1.9), 4.5 µm (gsd 1.8) and 4.1 µm (gsd 2.0).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Standard starting concentration for aerosols (OECD 436 Appendix 3c) when no information is available on inhalation toxicity.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Remarks on duration:
To compensate for interruptions (maximum 3 minutes per treatment), the generation time was elongated by maximum 3 minutes in order to achieve an actual exposure time of 240 minutes.
Concentrations:
The study was performed following a stepwise exposure scenario (Annex 3c in OECD 436). Three animals of each sex were exposed to a test item target concentration of 1 mg/L. Based on the results, one additional group of three animals of each sex was exposed to the technically maximum attainable concentration of 3.9 mg/L.
No. of animals per sex per dose:
3 animals of both sexes per exposure group. Two exposure groups. Females were nulliparous and non-pregnant.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Animals were checked for mortality, behavioral signs of distress and effects on respiration at least three times during exposure.
Post exposure observations were performed at periodic intervals on the day of exposure (at least two times) and once daily thereafter.
Animals were weighed individually on Day 1 (pre-exposure), 2, 4 and 8 and 15.
- Necropsy of survivors performed: yes
All moribund animals and animals surviving to the end of the observation period were sacrificed by an intra-peritoneal injection of Euthasol® (at least 250 mg/kg). All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.
- Other examinations performed: mortality, behavioral signs of distress, effects on respiration and body weight
Statistics:
No statistical analysis will be performed (the method used is not intended to allow the calculation of a precise LC50 value).
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 3.9 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortality occurred.
Clinical signs:
other: At 1 mg/L, shallow breathing labored respiration and restless behavior were seen during exposure (not presented in the table). After exposure, lethargy, hunched posture, labored respiration, slow breathing and rales were seen for the animals. The animal
Body weight:
Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
GHS criteria not met
Conclusions:
The inhalation LC50, 4h value of CH02906 in Wistar rats was established to exceed 3.9 mg/L.
Based on these results CH02906 does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).
Executive summary:

The objective of this study was to assess the acute inhalation toxicity of CH02906 in rats of both sexes in rats following a single 4 hour nose-only exposure to one or more defined concentrations. Animals were retained for a 14 day post-exposure observation period.

The study was carried out based on the guidelines described in:

·      Acute Inhalation Toxicity- Acute Toxic Class Method,September 2009.

CH02906 was administered as a dust by nose-only inhalation for 4 hours to two groups of three male and three female Wistar rats. Mortality and clinical signs were observed daily during the observation period and body weights were determined on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (Day 15).

 

At 1 mg/L, the time-weighted mean measured air concentration was 1.1 ± 0.02 mg/L.

At a target concentration of 5 mg/L, the time-weighted mean measured air concentration was 3.9 ± 0.09 mg/L. This concentration was found to be the technically maximum attainable concentration. The nominal concentration was 235 mg/L. The generation efficiency was 1.7%.

The air concentration was measured at several time points that were equally distributed over the exposure period, the results of which demonstrated that the item was sufficiently stable. Variation in measured air concentration was caused by adjustments to the generation equipment. By calculating the time-weighted mean measured air concentration, effects of these variations were taken into account resulting in a more accurate reflection of the mean exposure concentration over time.

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined at least twice during the exposure period. At 1 mg/L, the MMAD was 3.2 µm (gsd 1.9) and 3.3 µm (gsd 2.2). At 3.9 mg/L, the MMAD was 4.0 µm (gsd 1.9), 4.5 µm (gsd 1.8) and 4.1 µm (gsd 2.0). Agglomeration of aerosol particles at the high concentration might have resulted in the MMAD values to fall outside the recommended range of 1 - 4 µm. The MMAD only slightly exceeded this range and there was no evidence for item deposition in the upper airways. Since good distribution throughout the lung requires particles with an aerodynamic diameter between 1 and 5μm, it can be assumed that sufficient deposition in the lower respiratory tract occurred during the exposure.  

No mortality occurred.

At 1 mg/L, shallow breathing labored respiration and restless behavior were seen during exposure (not presented in the table). After exposure, lethargy, hunched posture, labored respiration, slow breathing and rales were seen for the animals. The animals had recovered from the clinical signs between Days 3 and 6.

At 3.9 mg/L, slow breathing was seen during exposure (not presented in the table). After exposure, lethargy, hunched posture, labored respiration, slow breathing, rales, piloerection and a dull right eye were seen for the animals. The animals had recovered from the clinical signs by Day 3.

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

No abnormalities were found at macroscopic post mortem examination of the animals.

 

The inhalation LC50, 4hvalue of CH02906 in Wistar rats was established to exceed 3.9 mg/L.

Based on these results CH02906 does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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:
03 Feb 2017 - 21 Feb 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1987
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
August 1998
Deviations:
no
Qualifier:
according to
Guideline:
other: EC No 440/2008 Part B. Acute Toxicity (Dermal)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to
Guideline:
other: Appendix to Director General Notification, No. 12-Nousan-8147. JMAFF
Version / remarks:
November 2000
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
Crl: WI(Han); Outbred, SPF-Quality
Sex:
male/female
Details on test animals and environmental conditions:
Test animals
Source:
Charles River France, L’Arbresle, France
Number of Animals:
5 males and 5 females (females were nulliparous and non-pregnant).
Age at the Initiation of Dosing:
Young adult animals (approximately 10 weeks old) were selected.
Weight at the Initiation of Dosing:
276 to 313 g (males) and 189 to 198 g (females).

Animal Identification
At study assignment, each animal was identified using a tail mark with indelible ink.

Environmental Acclimation
The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.

Housing
On arrival and following assignment to the study, animals were group housed (up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. These housing conditions were maintained unless deemed inappropriate by the Study Director and/or Clinical Veterinarian. The room(s) in which the animals were kept were documented in the study records.
Animals were separated during designated procedures/activities. Each cage was clearly labeled.

Environmental Conditions
Target temperatures of 18°C to 24°C with a relative target humidity of 40% to 70% were maintained. A 12‑hour light/12‑hour dark cycle was maintained, except when interrupted for designated procedures. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

Food
Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility.
It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.

Water
Municipal tap-water was freely available to each animal via water bottles.
Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility.
It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.

Animal Enrichment
For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.
Type of coverage:
occlusive
Vehicle:
propylene glycol
Remarks:
Trial preparations were performed at the Test Facility to select the suitable vehicle and to establish a suitable formulation procedure.
Details on dermal exposure:
Administration of Test item
A single dose of test item was administered to the appropriate animals by dermal application on Day 1. One day before dosing, an area of approximately 5x7 cm on the back of the animals was clipped. The test item was applied in an area of approximately 10% of the total body surface, i.e. approximately 25 cm² for males and 18 cm² for females. The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only. The application period was 24 hours, after which the dressing was removed and the skin cleaned of residual test item using water or an appropriate vehicle.
The dose level was 2000 mg/kg body weight.
The dose volume for each animal was based on the body weight measurement prior to dosing. A dose volume of 10 mL/kg body weight will be used for each dose.
The dosing formulations were stirred continuously during dose administration.
Justification of Route and Dose Levels
The dermal route was selected as it is a possible route of human exposure during manufacture, handling or use of the test item. The dose level was based on the OECD test guidelines.
Duration of exposure:
24 hours
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
Mortality/Moribundity Checks
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, once in the morning and once in the afternoon. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.

Clinical Observations
Postdose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days.
All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate), particular attention being paid to the animals during and for the first hour after dosing.
Animals were weighed individually on Day 1 (predose), 8 and 15.

Terminal Procedures
All animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
No mortality occurred
Clinical signs:
Chromodacryorrhoea (snout) was noted in the majority of animals on Days 1 and/or 2. Additionally, rales were noted for two female animals on Day 1 only.
Alopecia of the forelegs was noted for one female animal throughout the entire observation period, this effect was considered not indicative of toxicity.
General erythema, maculate erythema, scales and/or scabs were seen in the treated skin-area of the animals during the observation period. These local effects were considered not to have affected the conclusion of the study.
Body weight:
The mean body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
GHS criteria not met
Conclusions:
Based on these results, CH02906 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).
Executive summary:

The objective of this study was to determine the potential toxicity of CH02906, when given by a single dermal dose.

Assessment of Acute Dermal Toxicity with CH02906 in the Rat.

The study was carried out based on the guidelines described in:

·     OECD No. 402 (1987) "Acute Dermal Toxicity"

·     EC No 440/2008, B: "Acute Toxicity (Dermal)"

·     EPA, OPPTS 870.1200 (1998), "Acute Dermal Toxicity"

·     JMAFF Guidelines (2000), including the most recent revisions.

CH02906 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

 

No mortality occurred.

Chromodacryorrhoea[KV1] (snout) was noted in the majority of animals on Days 1 and/or 2. Additionally, rales were noted for two female animals on Day 1 only.

General erythema, maculate erythema, scales and/or scabswere seen in the treated skin-area of the animals during the observation period. These local effects were considered not to have affected the conclusion of the study.

The mean body weight gain during the observation period was within the range expected for rats used in this type of study.

No abnormalities were found at macroscopic post mortem examination of the animals.

 

The dermal LD50 value of CH02906 in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, CH02906 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

 [KV1]Chromodacryorrhea?

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Justification for classification or non-classification