Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: WoE: Based on the read-across approach from the analogue OS1600, the oral LD50 was determined to be 1457 mg/kg bw.
Acute oral toxicity: WoE: Acute oral toxicity: Based on the read-across approach from the analogue OS2600, the oral LD50 was determined to be between 1141 and 2282 mg/kg bw.
Acute oral toxicity: WoE: Acute toxicity oral: Based on the read-across approach from the analogue MPKO, the oral LD50 was determined to be 1457 mg/kg bw.
Acute dermal toxicity: Key study: Acute toxicity dermal: Based on the read-across approach from the analogue OS2200, the dermal LD50 was determined to be >1392 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
12 April -11 May 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The analogue OS2600 which shares the same functional group with OS9600, also has comparable values for the relevant molecular properties. The study has been conducted according to OECD guidance under GLPs.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Test material information:
Composition 1
Species:
rat
Strain:
other: Wistar strain, Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 8 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean (control group: 152 - 178 grams; 1000 mg/kg group: 147 - 177 grams ; 2000 mg/kg group: 153 - 179 grams).
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle.

IN-LIFE DATES: From: 12 April 2012 to 11 May 2012
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight.
Dose volume calculated as dose level (g/kg) / density (g/mL).

DOSAGE PREPARATION: The test substance was dosed undiluted as delivered by the sponsor. No correction for purity was made

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Based on anticipated mortality at the next higher dose.
Doses:
1000 mg/kg body weight
2000 mg/kg body weight


No. of animals per sex per dose:
1000 mg/kg: 6 (2 groups of three females in a stepwise manner)
2000 mg/kg: 3
Control animals:
yes
Details on study design:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 1000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups. A concurrent control group was added to the first two treated groups. Thereafter no concurrent control was added.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily. The time of death was recorded as precisely as possible.
Body weights: Days 1 (pre-administration), 8 and 15 and at death (if found dead after Day 1).
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: on Day 15
- Other examinations performed: bloodsampling on Day 5 and 15 (red blood cell count, reticutolcyte count, hemoglobin level and met-hemoglobin level, organ weights (spleen, kidney and liver) at necropsy, hostopathological examination of the spleen
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).
Sex:
female
Dose descriptor:
LD50
Effect level:
> 1 141 - < 2 282 mg/kg bw
Based on:
test mat.
Remarks on result:
other: (read-across from an analogue for which LD50 was 1000-2000 mg/kg bw)
Mortality:
No mortality occurred at 1000 mg/kg and control groups. Two females at 2000 mg/kg (2/3) were found dead on Days 1 or 4.
Clinical signs:
No clinical signs were noted in the control animals.
Animals at 1000 mg/kg showed lethargy, hunched posture, uncoordinated movements, piloerection, ptosis, slow breathing, and/or watery discharge from the eye on Day 1.
Animals at 2000 mg/kg showed lethargy, flat and/or hunched posture, uncoordinated movements, slow breathing, piloerection, hypothermia and/or ptosis until death (two animals) or on Days 1-3 (surviving animal).
Body weight:
The body weight gain shown by the surviving animals over the study period was considered to be similar to the control animals.
Gross pathology:
Necropsy did not reveal any toxicologically relevant alterations.
A control animal showed pelvic dilation and black nodules in the liver, which are considered incidental findings not related to the test substance. One animal at 2000 mg/kg showed advanced autolysis and the skin of the left region of the head was missing due to cannibalism.
Other findings:
The animals at 1000 mg/kg showed lower red blood cell counts in combination with higher percentage of reticulocytes, compared to the control animals, on Day 5 and 15. A lower level of total hemoglobin was noted on Day 5, which recovered on Day 15. The slight higher met-hemoglobin levels on Day 5 and/or 15 (as compared to the controls) are within the range to be expected for rats of this age and strain.
The surviving animal treated at 2000 mg/kg showed comparable trends in all four parameters.

Liver, spleen and kidney weights of treated animals were considered to be similar to those of control animals.

There were no treatment-related findings in the spleens of the 1000 mg/kg treated rats.
No direct cause of death at 2000 mg/kg could be established from the examined sections of the spleen.
There was a treatment related microscopic finding in the 2000 mg/kg treated rats. Atrophy of the white and red pulp of the spleen was noted at minimal degree in the one surviving female and at moderate degree in one female (found dead on Day 4).
The remainder of the recorded microscopic findings in the spleen were all within the normal range of background alterations encountered in rats of this age and strain and therefore not regarded as test article related.

The data matrix is included in the reporting format attached in Section 13.

The results from the read-across approach are expressed (calculated) on the basis of the molecular weight of OS9600 and OS2600 since the basic structures of the target and source substances are the same.

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the read-across approach from the analogue OS2600, the oral LD50 for OS9600 was determined to be between 1141 and 2282 mg/kg bw .
Executive summary:

The acute oral toxicity study was performed on the analogue OS2600 in accordance with OECD Guideline 423 and EU Method B1 tris (GLP study). Initially, 1000 mg/kg bw test item was administered by oral gavage to 3 female Wistar rats. In a stepwise procedure additional groups of females were dosed at 1000 and 2000 mg/kg body weight. Concurrent control groups were added in the first two treatment groups. All animals were subjected to daily observations and weekly determination of body weight. Blood samples were taken from all surviving animals on Day 5 and prior to necropsy. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15). Histopathology examination was performed on the spleen. No mortality occurred at 1000 mg/kg and control groups. Two females at 2000 mg/kg (2/3) were found dead on Days 1 or 4 and therefore, the oral LD50 value of OS 2600 in Wistar rats was established to be within the range of 1000-2000 mg/kg body weight. Haematology parameters were affected by treatment at a dose level of 1000 mg/kg, and the same trend was noted at 2000 mg/kg. The following findings were included: lower red blood cell count, higher reticulocyte percentage and lower haemoglobin levels. These findings were not accompanied by treatment related morphological changes in the spleen at 1000 mg/kg. Two animals at 2000 mg/kg (one surviving and one which died on Day 4) showed atrophy of the white and red pulp of the spleen at microscopic examination. Based on these results, the read-across was applied and the oral LD50 for OS9600 was determined to be between 1141 and 2282 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
1 143 mg/kg bw
Quality of whole database:
The study is a GLP compliant and has a Klimisch score 2 (read-across from an endpoint with Klimisch score 1)

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The analogue OS2200 which shares the same functional group with OS9600, also has comparable values for the relevant molecular properties for acute dermal toxicity. The study has been conducted according to OECD guidance under GLPs.
Reference:
Composition 0
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Olac Ltd., Bicester, Oxon, England
- Weight at study initiation: 214 to 238 g
- Housing: Individually in metal cages with wire mesh floors
- Diet (e.g. ad libitum): Ad libitum (SDS LAD 1)
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30 - 70% RH
- Air changes (per hr): 10 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours of artificial light (0700 - 1900 hours) in each 24-hour period
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: dorso-lumbar region (one day prior to treatment, hair was clipped), 10% of the total body surface. The test substance was applied by spreading it evenly over the prepared skin.
- % coverage, type of wrap: The treated area (approximately 50 mm x 50 mm) was then promptly covered with gauze which was held in place with a non-irritative dressing encircled firmly around the trunk.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): warm (30- 40°C) water and blotted dry with absorbent paper
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.1 mL/kg bw (2000 mg/kg bw)
Duration of exposure:
24 hours
Doses:
1
No. of animals per sex per dose:
5 rats per sex and dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: mortality and clinical signs: twice daily; body weight: Days 1, 8 and 15.
- Necropsy of survivors performed: Yes. All animals were killed on Day 15 by carbon dioxide asphyxiation and were subjected to a macroscopic examination which consisted of opening the abdominal and thoracic cavities. The macroscopic appearance of all examined tissues was recorded.
- Other examinations performed: clinical signs, body weight, macroscopic examination, other: Local dermal irritation at the treatment site was assessed daily and scored (Draize scoring method)
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 392 mg/kg bw
Based on:
test mat.
Remarks on result:
other: (read-across from an analogue for which LD50 > 2000 mg/kg bw)
Mortality:
There were no deaths.
Clinical signs:
There were no signs of systemic reaction to treatment.
Body weight:
In comparison with historical data from the Department of Industrial Toxicology of HRC, slightly low bodyweight gains were recorded for two males and two females on Day 8. All other rats achieved satisfactory bodyweight gains throughout the study.
Gross pathology:
No macroscopic abnormalities were observed for animals killed on Day 15.
Other findings:
Sites of application showed no irritation or other dermal changes.

The data matrix is included in the reporting format attached in Section 13.

The results from the read-across approach are expressed (calculated) on the basis of the molecular weight of OS9600 and OS2200 since the basic structures of the target and source substances are the same.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the read-across approach from the analogue substance OS2200, the acute lethal dermal dose of OS9600 to rats was determined to be greater than 1392 mg/kg bw.
Executive summary:

An acute dermal toxicity study was performed on the analogue substance OS2200 according to EU Method B.3. A group of ten rats (five males and five females) was given a single dermal application of the test substance, as supplied, at a dose level of 2000 mg/kg bw. All animals were killed and examined macroscopically on Day 15, the end of the observation period. There were no deaths and no signs of systemic reaction to treatment. Sites of application showed no irritation or other dermal changes. In comparison with historical data from the Department of Industrial Toxicology of HRC, slightly low bodyweight gains were recorded for one male and three females on Day 8. All other rats achieved satisfactory bodyweight gains throughout the study. No abnormalities were recorded at the macroscopic examination on Day 15. The LD50 of the analogue OS220 was therefore > 2000 mg/kg bw. Based on these results, the read-across approach was applied and the acute lethal dermal dose of OS9600 to rats was determined to be > 1392 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
1 392 mg/kg bw
Quality of whole database:
The study is a GLP compliant and has a Klimisch score 2 (read-across from an endpoint with Klimisch score 1)

Additional information

Acute oral toxicity:

Weight of evidence: Read-across approach from experimental data on the analogue OS1600:

An acute oral toxicity study was performed on the analogue substance OS1600 according to OECD Guideline 425 (GLP study). Based on the obtained results (LD50 ca. 1234 mg/kg bw, hematological effects observed), the read-across approach was applied and the oral LD50 in rats for the substance OS9600 was determined to be ca 1457 mg/kg bw.

Weight of evidence: Read-across approach from experimental data on the analogue OS2600:

The acute oral toxicity study was performed on the analogue OS2600 in accordance with OECD Guideline 423 (GLP study). Based on the obtained results (LD50 within the range of 1000-2000 mg/kg bw and haematology parameters affected by treatment at 1000 mg/kg and 2000 mg/kg), the read-across approach was applied and the oral LD50 in rats for the substance OS9600 was determined to be between 1141 and 2282 mg/kg bw.

Weight of evidence: Read-across approach from experimental data on the analogue MPKO:

An acute oral toxicity study was performed with MPKO, the hydrolysis product of OS9600, in the rat in accordance with OECD Guideline 425 (GLP study). The LD50 in rats was determined to be ca. 1133 mg/kg bw and within the range of 300 -2000 mg/kg bw. Effects on haematology parameters were observed. Based on the read-across approach, the oral LD50 in rats for OS9600 was estimated to be 1514 mg/kg bw.

Acute dermal toxicity:

Weight of evidence: Read-across approach from experimental data on the analogue OS2200:

An acute dermal toxicity study was performed on the analogue substance OS2200 according to EU Method B.3 (GLP study). Based on the obtained results (LD50 > 2000 mg/kg bw, no adverse effects observed at the highest dose tested), the read-across approach was applied and the dermal LD50 in rats for the substance OS9600 was determined to be > 1392 mg/kg bw.


Justification for selection of acute toxicity – oral endpoint
The lowest LD50 value was chosen.

Justification for selection of acute toxicity – inhalation endpoint
According to REACH Annex VIII, column 2: In addition to the oral route, for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. If there is only one route of exposure, information for only that route need be provided. The information is provided for dermal route.

Justification for selection of acute toxicity – dermal endpoint
Only one study available.

Justification for classification or non-classification

Based on the available data, the substance is classified as Acute Toxicity Category 4, H302 in accordance with CLP Regulation (EC) no. 1272/2008 since the acute toxicity value (oral) expressed as LD50 is between 300 and 2000 mg/kg bw.