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Administrative data

Description of key information

Acute oral toxicity > 5000 mg/kg (substance)
Acute dermal toxicity > 2000 mg/kg (read across to structural analogue)

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:
July 14, 2014 to August 4, 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed in accordance with OECD test guideline and in compliance with GLP.
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
other: CD® [Crl:CD®(SD)]
Sex:
female
Details on test animals and environmental conditions:
Species: RatStrain: CD® [Crl:CD®(SD)]Source: Charles River LaboratoriesExpected Age: 7 weeks of age at arrivalExpected Weight at Arrival: Females weighed 100 to 170 g, as measured within 3 days of arrival. Number Ordered Female: 10Number on Study Female: Up to 9AcclimationDuration: At least 1 weekAcclimation Details: During this acclimation period, all animals were observed daily for any clinical signs of disease, and all animals were given a detailed clinical examination prior to selection for study.Selection for StudyBody Weight Range: ±20% of the mean body weightRandomization: Into treatment groups using a simple randomization procedure. All animals with any evidence of disease or physical abnormalities were not selected for study.Method of Identification: Each animal was assigned an animal number to be used in Provantis™ and was implanted with a microchip bearing a unique identification number. The individual animal number, implant number, and the MPI Research study number comprises a unique identification for each animal. The animal cage was identified by the study number, animal number, group number, and sex.Housing: IndividuallyCaging: Solid bottom cages with nonaromatic bedding. The bedding was sourced from an approved supplier and documented in the study data.Temperature: 68 to 79 °FHumidity: 30 to 70%Lighting: Fluorescent lighting was provided via an automatic timer for approximately 12 hours per day. On occasion, the dark cycle could be interrupted intermittently due to study-related activities.Water: Tap water was supplied ad libitum to all animals via an automatic water system unless otherwise indicated.Diet: The basal diet was block Lab Diet® Certified Rodent Diet #5002, PMI Nutrition International, Inc. This diet was available ad libitum unless designated otherwise. Each lot number used was identified in the study records.Water and Diet Contaminants: There are no known contaminants in the food or water that would interfere with this study. The drinking water used is monitored for specified contaminants at periodic intervals according to MPI Research SOP.Supplemental Enrichment: Animal enrichment was provided as described in MPI Research SOP.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Preparation: Test article was administered neat.Frequency: Day of administrationStorage Conditions: Room temperature
Doses:
Sighting Study: 2000 mg/kg. As no toxicity was noted an additional group was added at 5000 mg/kg.Main Study: 5000 mg/kg
No. of animals per sex per dose:
1 female per dose in the Sighting study4 females in the main study
Control animals:
no
Details on study design:
The 2000 mg/kg dose is specified by regulatory agencies for the limit test. Any additional doses were selected to estimate the LD50.The test article was administered once on Day 1 via oral gavage to one animal. The initial dose level for the sighting study was 2000 mg/kg. Due to there being no toxicity noted, an additional animal was administered the test article at a dose level of 5000 mg/kg. The results of the sighting study determined that 5000 mg/kg was the starting dose for the main study animals. Individual doses were based on the most recent body weight measurements. The test article was administered at a dose volume of 2.08 mL/kg to the initial animal dosed in the sighting study and at a dose volume of 5.21 mL/kg for the second animal in the sighting study and for all main study animals. The animals were fasted overnight prior to test article administration and had food returned following the completion of dosing.Observations for morbidity, mortality, injury, and the availability of food and water were conducted twice daily for all animals. Observations for clinical signs were conducted on Day 1 at 30 minutes, 2, and 4 hours postdose, and daily thereafter. Body weights were measured and recorded prior to dosing on Day 1 and weekly thereafter.At study termination (Day 15), the animals were euthanized by carbon dioxide inhalation. Euthanasia was confirmed via exsanguination via severing the vena cava. Necropsy examinations were performed under procedures approved by a veterinary pathologist on all animals euthanized at the scheduled necropsy. The animals were examined carefully for external abnormalities including palpable masses. The skin was reflected from a ventral midline incision, any subcutaneous masses were identified and correlated with antemortem findings, and the abdominal, thoracic, and cranial cavities were examined. Following the evaluation, the carcasses were discarded and no tissues were saved.
Statistics:
No statistical analyses are required or will be performed for this study.
Preliminary study:
The test article was administered once on Day 1 via oral gavage to one animal. The initial dose level for the sighting study was 2000 mg/kg. Due to there being no toxicity noted, an additional animal was administered the test article at a dose level of 5000 mg/kg. The results of the sighting study determined that 5000 mg/kg was the starting dose for the main study animals.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals survived to the scheduled necropsy.
Clinical signs:
No adverse clinical findings indicative of systemic toxicity were noted. Yellow discoloured hair was noted in the anogenital region on Day 1 at 4 hours postdose for animal number 8002. This is a common finding in an animal of this age, sex, and strain, thus deemed not test article-related.
Body weight:
There were no adverse test article-related effects on body weight. Body weights were within normal range for this species.
Gross pathology:
The tissues of all animals examined at necropsy were within normal limits.
Other findings:
No further findings detailed in the study report.

Record of Animal Fate and Disposition – Sighting Study - FEMALE

Group,

Animal Number

Fate

Day

2000 mg/kg

8001

 

Terminal necropsy

 

15

5000 mg/kg

8002

 

Terminal necropsy

 

15

 

Record of Animal Fate and Disposition – Main Study - FEMALE

Group,

Animal Number

Fate

Day

5000 mg/kg

8003

8004

8005

8006

 

Terminal necropsy

Terminal necropsy

Terminal necropsy

Terminal necropsy

 

15

15

15

15

 

Individual Detailed Clinical Observations – Sighting Study – FEMALE

30 minute & 2 hour post dose

Group,

Animal Number

Day(s) Sign Present

2000 mg/kg

8001      No abnormalities detected

 

1

5000 mg/kg

8002      No abnormalities detected

 

1

 

Individual Detailed Clinical Observations – Sighting Study – FEMALE

4 hour post dose

Group,

Animal Number

Day(s) Sign Present

2000 mg/kg

8001     No abnormalities detected

 

1

5000 mg/kg

8002      Hair discoloured, Yellow, Anogenital region

 

1

 

Individual Detailed Clinical Observations – Sighting Study – FEMALE

Group,

Animal Number

Day(s) Sign Present

2000 mg/kg

8001      No abnormalities detected

 

2 - 14

5000 mg/kg

8002      No abnormalities detected

 

2- 14

 

Individual Detailed Clinical Observations – Main Study – FEMALE

30 minute, 2 and 4 hour post dose

Group,

Animal Number

Day(s) Sign Present

5000 mg/kg

8003      No abnormalities detected

8004      No abnormalities detected

8005      No abnormalities detected

8006      No abnormalities detected

 

1

1

1

1

 

Individual Detailed Clinical Observations – Main Study – FEMALE

Group,

Animal Number

Day(s) Sign Present

5000 mg/kg

8003      No abnormalities detected

8004      No abnormalities detected

8005      No abnormalities detected

8006      No abnormalities detected

 

2 – 14

2 – 14

2 – 14

2 – 14

 

Individual Body Weight Values – Sighting Study, g - FEMALE

Group,

Animal Number

Study Interval (Week)

1

2

3

2000 mg/kg

8001

 

176

 

218

 

248

5000 mg/kg

8002

 

167

 

185

 

199

 

Individual Body Weight Values – Main Study, g - FEMALE

Group,

Animal Number

Study Interval (Week)

1

2

3

5000 mg/kg

8003

8004

8005

8006

 

194

205

196

192

 

217

240

225

217

 

227

269

236

234

 

Individual Animal Data Record: Pathology – Sighting Study, FEMALE

Terminal

Group,

Animal Number

 

Fate

Tissue

Observations

2000 mg/kg

8001

 

S

Macroscopic

All tissues

 

- within normal limits

5000 mg/kg

8002

 

S

Macroscopic

All tissues

 

- within normal limits

S – Scheduled necropsy

 

Individual Animal Data Record: Pathology – Main Study - FEMALE

Group,

Animal Number

 

Fate

 

Tissue

 

Observations

5000 mg/kg

8003

S

Macroscopic

All tissues

 

- within normal limits

8004

S

Macroscopic

All tissues

 

- within normal limits

8005

S

Macroscopic

All tissues

 

- within normal limits

8006

S

Macroscopic

All tissues

 

- within normal limits

S – Scheduled necropsy

Interpretation of results:
study cannot be used for classification
Remarks:
Migrated information
Conclusions:
Under the conditions of this study, where Hatcol 3346, was administered as a single oral dose to rats, the LD50 was greater than 5000 mg/kg.
Executive summary:

This study was conducted for the sponsor to evaluate the potential oral acute toxicity of the test article, Hatcol 3346. This study was conducted in accordance with Standard Operating Procedures (SOPs) and the protocol as approved by the Sponsor. No protocol or SOP deviations occurred. This study was based on the Organization for Economic Cooperation and Development (OECD) Guideline 420 (2001) and the Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Academy Press, Washington, D.C., 2011.

 

The 2000 mg/kg dose is specified by regulatory agencies for the limit test. Any additional doses were selected to estimate the LD50.

The test article was administered once on Day 1 via oral gavage to one animal. The initial dose level for the sighting study was 2000 mg/kg. Due to there being no toxicity noted, an additional animal was administered the test article at a dose level of 5000 mg/kg. The results of the sighting study determined that 5000 mg/kg was the starting dose for the main study animals. Individual doses were based on the most recent body weight measurements. The test article was administered at a dose volume of 2.08 mL/kg to the initial animal dosed in the sighting study and at a dose volume of 5.21 mL/kg for the second animal in the sighting study and for all main study animals. The animals were fasted overnight prior to test article administration and had food returned following the completion of dosing.

Observations for morbidity, mortality, injury, and the availability of food and water were conducted twice daily for all animals. Observations for clinical signs were conducted on Day 1 at 30 minutes, 2, and 4 hours postdose, and daily thereafter. Body weights were measured and recorded prior to dosing on Day 1 and weekly thereafter.

At study termination (Day 15), the animals were euthanized by carbon dioxide inhalation. Euthanasia was confirmed via exsanguination via severing the vena cava. Necropsy examinations were performed under procedures approved by a veterinary pathologist on all animals euthanized at the scheduled necropsy. The animals were examined carefully for external abnormalities including palpable masses. The skin was reflected from a ventral midline incision, any subcutaneous masses were identified and correlated with antemortem findings, and the abdominal, thoracic, and cranial cavities were examined. Following the evaluation, the carcasses were discarded and no tissues were saved.

 

All animals survived to the scheduled necropsy.

No adverse clinical findings indicative of systemic toxicity were noted. Yellow discoloured hair was noted in the anogenital region on Day 1 at 4 hours postdose for animal number 8002. This is a common finding in an animal of this age, sex, and strain, thus deemed not test article-related.

There were no adverse test article-related effects on body weight. Body weights were within normal range for this species.

The tissues of all animals examined at necropsy were within normal limits.

 

Under the conditions of this study, where Hatcol 3346, was administered as a single oral dose to rats, the LD50 was greater than 5000 mg/kg. No classification is applicable.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Recent study performed in accordance with OECD test guideline and in compliance with GLP.

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
Study period:
02 July 2003- 16 July 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to OECD test guidance in compliance with GLP and reported with a valid GLP certificate. The study is read across to an analogous substance; refer to image and further information below.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Species:Rat, Wistar strain Crl:(WI) BR (outbred, SPF-Quality). Recognised by international guidelines as the recommended test system (e.g.OECD, EC)Source: Charles River Deutschland, Sulzfeld, Germany.Number of animals: 5 males and 5 females (females were nulliparous and nonpregnant).Age and bodyweight: Young adult animals (approx. 9-10 weeks old) were selected. Bodyweight variation did not exceed +/- 20% of the sex mean.Identification: EarmarkConditionsAnimals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0:i 3.0°C (actual range: 17.2 - 23.7°C), a relative humidity of 30-70% (actual range: 44 - 83%) and 12 hours artificial fluorescent light and 12 hours darkness per day. Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relativehumidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.AccommodationIndividually housed in labelled Macrolon cages (type IIi, height 15 cm.) containing purified Sawdust as bedding material (SAWI, Jelu Werk, Rosenberg, Germany). Certificates of analysis were examined and then retained in the NOTOX archives. Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.DietFree access to standard pelleted laboratory animal diet (from Altromin (code VRF 1), Lage, Germany). Cer!ifcates of analysis were examined and then retained in the NOTOX archives.WaterFree access to tap-water. Certifcates of quarterly analysis were examined and then retained in the NOTOX archives.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Method: Dermal application.Clipping: One day before exposure (day -1) an area of approximately 5x7cm on the back of the animal was clipped.Application: The test substance was applied in an area of approx. 10% of thetotal body surface, i.e. approx. 25 cm2 for males and 18 cm2 for females. The test substance was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1 D), successively covered with aluminium foil and Coban flexible bandage'. A piece of Micropore tape was additionally used forfixation of the bandages in females only.Frequency: Single dosage, on day 1.Application period: 24 hours, after which dressings were removed and the skincleaned of residual test substance using water.
Duration of exposure:
24 h
Doses:
2000 mglkg (2.061 ml/kg) body weight. Dose volume calculated as follows: dose level : specific gravity.
No. of animals per sex per dose:
5 male, 5 female
Control animals:
no
Details on study design:
Mortality/Viability: Twice daily.Body weights: Days 1 (pre-administration), 8 and 15.Clinical signs: At periodic intervals on the day of dosing (day 1) and once dailythereafter, until day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales: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: At the end of the observation period, all animals were sacrificed byasphyxiation using an oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
No statistical analysis was performed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Clinical signs:
Hunched posture and/or chromodacryorrhoea were noted among all animals on day 2. In addition, maculate erythema or scales were seen on the treated skin site of one male and female between days 3 and 7.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mor!em examination of the animals.
Interpretation of results:
not classified
Remarks:
Migrated informationCriteria used for interpretation of results: EU
Conclusions:
Read across to structural analogue. Structural details are listed above. No mortality occurred.Hunched posture and/or chromodacryorrhoea were noted among all animals on day 2. In addition, maculate erythema or scales were seen on the treated skin site of one male and female between days 3 and 7.The 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 HATCOL 3331 in Wistar rats was established to exceed 2000 mg/kg body weight.
Executive summary:

Read across to structural analogue. Structural details are listed above.

No mortality occurred.

Hunched posture and/or chromodacryorrhoea were noted among all animals on day 2. In addition, maculate erythema or scales were seen on the treated skin site of one male and female between days 3 and 7.

The 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 HATCOL 3331 in Wistar rats was established to exceed 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
OECD GLP Study performed on read across substance. Substances are chemically equivalent.

Additional information

No clinical signs of toxicity and with regard to gross pathology no signs of systemic toxicity and no abnormalities were observed. The acute toxicity of the substance is summarised as follows:

Oral

All animals survived to the scheduled necropsy.

No adverse clinical findings indicative of systemic toxicity were noted. Yellow discoloured hair was noted in the anogenital region on Day 1 at 4 hours postdose for animal number 8002. This is a common finding in an animal of this age, sex, and strain, thus deemed not test article-related.

There were no adverse test article-related effects on body weight. Body weights were within normal range for this species.

The tissues of all animals examined at necropsy were within normal limits.

 

Under the conditions of this study, where Hatcol 3346, was administered as a single oral dose to rats, the LD50 was greater than 5000 mg/kg. No classification is applicable.

Inhalation.

In accordance with column 2, adaptation of Annex VIII (section 8.5.2) testing by the inhalation route is deemed inappropriate as exposure to humans via inhalation is unlikely to occur as the substance does not have a high vapour pressure, and will not be used in a manner likely to produce significant aerosols or droplets of an inhalable size. The dermal route is considered to be a more appropriate second route of exposure, and sufficient data is available to address this endpoint.

Dermal

Read across to structural analogue.

No mortality occurred.

Hunched posture and/or chromodacryorrhoea were noted among all animals on day 2. In addition, maculate erythema or scales were seen on the treated skin site of one male and female between days 3 and 7. The 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 HATCOL 3331 in Wistar rats was established to exceed 2000 mg/kg body weight.


Justification for selection of acute toxicity – oral endpoint
Recent study performed in accordance with OECD test guideline and in compliance with GLP.

Justification for selection of acute toxicity – inhalation endpoint
In accordance with column 2, adaptation of Annex VIII (section 8.5.2) testing by the inhalation route is deemed inappropriate as exposure to humans via inhalation is unlikely to occur as the substance does not have a high vapour pressure, and will not be used in a manner likely to produce significant aerosols or droplets of an inhalable size. The dermal route is considered to be a more appropriate second route of exposure, and sufficient data is available to address this endpoint.

Justification for selection of acute toxicity – dermal endpoint
OECD GLP Study performed on read across substance. Substances are chemically equivalent.

Justification for classification or non-classification

Based on the above mentioned result, classification according to the CLP Regulation (EC) 1272/2008 is not necessary.