Registration Dossier

Administrative data

Description of key information

Acute toxicity - oral: In a K1 acute oral toxicity study (up-and down procedure) in Sprague-Dawley rats, performed according to OECD Guideline 425 (acute toxic class method) and EPA OPPTS 870.1100 - Acute Oral Toxicity, an LD50 value of 1000 mg/kg bw was observed (Vasquez, 2012).
Acute toxicity - dermal: In a K1 acute dermal toxicity study in Sprague Dawley rats, performed according to OECD Guideline 402, EPA OPPTS 870.1200, and EEC Method B.3, an LD50 value greater than 2000 mg/kg bw was observed (Vasquez, 2011).

Acute toxicity - inhalation: No acute inhalation toxicity study is performed.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2011-12-05 to 2011-12-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
yes
Remarks:
relative humidity 18 to 61%, 1 animal not weighed at time of death
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): XTJ-568
- Substance type: Clear colorless liquid
- Physical state: liquid
- Analytical purity: 97% (primary amine)
- Composition of test material, percentage of components: water (0.03%), acetylatable (9.2 meq/g), total amine (8.7 meq/g)
- Lot/batch No.: 0G704
- Stability under test conditions: no data
- Storage condition of test material: 21 - 26.7°C
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan
- Age at study initiation: 9 to 10 weeks at start of dosing; records of dates of birth for animals used in this study are retained in the Calvert archives.
- Weight at study initiation: 191 - 202 grams
- Fasting period before study: Animals were fasted overnight prior to dose administration.
- Housing: Animals were group housed by sex upon receipt and individually housed upon assignment to study in compliance with the National Research Council " Guide for the Care and Use of Laboratory Animals". Calvert is a USDA registered and fully AAALAC accredited facility. The room in which the animals were kept was documented in the study records. no other species were kept in the same room.
- Diet (e.g. ad libitum): ad libitum with fasting overnight prior to dose administration
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days prior to dosing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 22°C
- Humidity (%): 18 to 61%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Dose preparation: An initial preparation of the test article in distilled water resulted in an undoseable suspension with a pH of 13. Therefore, the proper amount of the test article was measured according to volume since it was a liquid, and was allocated/dosed neat as received from the Sponsor with a pH of 10. The stock bottle was inverted several times prior to dispensing.

Dose volumes (specific density 1 g/ml): 0.32, 0.55, 1 and 2 ml/kg.
Doses:
320, 550, 1000 or 2000 mg/kg
No. of animals per sex per dose:
A total of six rats received the substance.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
mortality/morbidity: manually recorded once daily
clinical observations: clinical observations were collected manually. On the day of dosing (Day 1) animals were observed prior to dosing and approximately 30 minutes and 4 hours post-dose. Animals were observed once daily thereafter (Days 2-15).
body weight: Animal weights were manually recorded prior to dosing on Day 1, and on Day 8 and 15, or upon death.
- Necropsy of survivors performed: yes. The necropsy included examination of: the external body surface, all orifices, the cranial, thoracic and abdominal cavities and their contents. All surviving animals were euthanized by CO2 asphyxiation.
Statistics:
The LD50 was calculated for the main test using AOT425StatPgm developed by Westat May, 2001.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
1 000 mg/kg bw
Based on:
test mat.
95% CL:
>= 275.7 - <= 2 580
Mortality:
Mortality was not observed in any of the animals dosed at 320 or 550 mg/kg of the substance. Two of three animals receiving the test article at 1000 mg/kg were found dead as well as the single animal dosed at 2000 mg/kg.
Clinical signs:
Clinical observations observed included piloerection at 30 minutes in the single animal at 320 mg/kg and in the surviving animal at 1000 mg/kg. no clinical signs were observed prior to death in the two other animals at 1000 mg/kg. Piloerection, decreased activity, decreased body tone and abnormal gait and stance were observed at 4 hours in the animal treated at 550 mg/kg. At 2000 mg/kg, decreased body tone and abnormal gait were observed at 30 minutes and death was observed at 4 hours.
Body weight:
No biologically significant effect was seen on body weights of the surviving animals on Days 8 and 15.
Gross pathology:
Terminal necropsy revealed no visible lesions in the animals at 320, 550 or the surviving animal at 1000 mg/kg. Necropsy of two animals found dead at 1000 and 2000 mg/kg revealed dark red fluid-filled intestines. The third animal found dead had no visible lesions.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Based on the results of this study, the oral LD50 for the substance in rats was estimated to be 1000 mg/kg (95% PL Confidence interval of 275.7 to 2580 mg/kg). The substance is considered classified as acute oral toxicant category 4 according to the criteria laid down in the CLP Regulation (EC) 1272/2008.
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2003-01 to 2003-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): XTJ 568
- Physical state: Clear colourless liquid
- Analytical purity: 97.8%
- Impurities (identity and concentrations): no data
- Purity test date: no data
- Lot/batch No.: 8191-34
- Expiration date of the lot/batch: 2004-01-01
- Stability under test conditions: no data
- Storage condition of test material: At room temperature in the dark
- Other: specific gravity: > 0.94 g/cm3
Species:
rat
Strain:
Wistar
Remarks:
Wistar strain Crl:(WI) BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfield, Germany
- Age at study initiation: approx. 8-9 weeks old
- Weight at study initiation: females 199 +/- 7 g (200 mg/kg dose); males 258 +/- 11 g (200 mg/kg dose); females 175 +/-7 g (2000 mg/kg dose)
- Fasting period before study: food was withheld overnight (for a maximum of 20 hours) prior to dosing until 3-4 hours after administration of the test substance
- Housing: Animals were housed in a controlled environment. Group housing of 3 animals per sex per cage labelled Macrolon cages (type IV; height 18 cm) containing purified sawdust as bedding material (SAWI, Jelu Werk, Rosenberg, Germany).
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.0 - 24.3 °C
- Humidity (%): 52 - 80%
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Vehicle preparation: The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the test substance. The concentration was made for specific gravity of the test substance. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of ml/kg body weight
- Justification for choice of vehicle: The vehicle formulation was selected based on trial formulations performed at NOTOX


Doses:
200 mg/kg (10 ml/kg) body weight
2000 mg/kg (10 ml/kg) body weight
No. of animals per sex per dose:
Each dose group consisted of 3 animals of one sex (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: mortality/viability (twice daily); 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: yes: the animals surviving to the end of the observation period were sacrificed by asphyxiation using an oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded
Statistics:
No statistical analysis was performed.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
200 - 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Male: 200 mg/kg bw; Number of animals: 3; Number of deaths: 0
Female: 200 mg/kg bw; Number of animals: 3; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 3; Number of deaths: 2
Clinical signs:
Signs of toxicity related to dose levels:
All animals dosed at 200 mg/kg body weight displayed hunched posture and/or piloerection on days 1 and/or 2. Lethargy was additionally noted in all males at 200 mg/kg body weight on day 1. Females dosed at 2000 mg/kg body weight showed hunched posture, piloerection, lethargy uncoordinated movements, ptosis, hypothermia, shallow respiration and/or chromodacryorrhoea. The surviving female at 2000 mg/kg bw had recovered from the symptoms by day 5.

posture and/or piloerection on days 1 and/or 2. Lethargy
was additionally noted in all males at 200 mg/kg body weight

on day 1. Females dosed at 2000 mg/kg body weight showed
hunched posture, piloerection, lethargy uncoordinated
movements, ptosis, hypothermia, shallow respiration and/or
chromodacryorrhoea

The surviving female at 2000 mg/kg body weight had recovered

from the symptoms by day 5.


The mean body weight gain shown by the surviving animals
over the study period was considered to be normal. Reduced
body weight gain was noted for one female dosed at 200 mg/kg

body weight between days 8 and 15.
Body weight:
The mean body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain. Reduced body weight gain was noted for one female dosed at 200 mg/kg body weight between days 8 and 15.
Gross pathology:
Effects on organs: Females found dead after treatment displayed dark red discolouration or foci on the glandular mucosa of the stomach. One of these females showed signs of beginning autolysis, and fluid in the uterus and dark red foci were noted on the thymus. No abnormalities were found at macroscopic examination of the other animals.
Other findings:
No data
Interpretation of results:
study cannot be used for classification
Conclusions:
The oral LD50 value of the test substance in Wistar rats was established to be within the range of 200-2000 mg/kg body weight.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because the substance is classified as corrosive to the skin
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:
experimental study
Adequacy of study:
key study
Study period:
2011-06-05 - 2011-06-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
- Name of test material (as cited in study report): XTJ-568
- Substance type: Clear colorless liquid
- Physical state: Liquid
- Lot/batch No.: 0G704
- Storage conditions: Room temperature, 21-27°C
- Purity: A Certificate of Analysis was not provided with this study. However, the same batch was used for the acute oral toxicity study (OECD 425), where a certificate of analysis was available and indicated a purity (primary amine) of 97%.
- Other: No stability data on bulk test material were available.
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan
- Age at study initiation: 12 to 14 weeks; records of dates of birth for animals used in this study are retained in the Calvert archives
- Weight at study initiation: males: 345-382 grams; females: 223-268 grams
- Fasting period before study: No
- Housing: Animals were group housed by sex upon receipt and individually housed upon assignment to study in compliance with the National Research Council "Guide for the Care and Use of Laboratory Animals." Calvert is a USDA registered and fully AAALAC accredited facility. The room in which the animals were kept was documented in the study records. No other species were kept in the same room.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: for a minimum for 5 days prior to dosing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 25°C
- Humidity (%): 29 to 74%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES: From: 5 June 2011 To: 20 June 2011
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Dorsal area of the trunk of the test animals
- % coverage: >= 10%
- Type of wrap if used: The test article was applied on an intact skin site for each animal, covered with a gauze patch/dental dam, wrapped with an elastic bandage and secured with non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Post-exposure, the site was unwrapped and wiped appropriately (with gauze and water) to remove residual test article.

TEST MATERIAL
- Dosage levels were calculated on a mg/kg basis for each individual animal based on body weight. For the liquid test article, the dose was adjusted for test article relative density (0.94 g/mL).
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical observations: frequency: clinical observations were recorded immediately after unwrap and daily thereafter through Day 15. At the time of clinical observations, the dose site was also evaluated for dermal irritation.
Body weight: frequency: Animals were weighed prior to dosing on Day 1 and on Days 8 and 15 or upon death.
- Necropsy of survivors performed: yes: A gross necropsy was performed by Calvert peronnel at study termination or when animals were found dead. The necropsy included examination of:
the exernal body surface
all orifices
the thoracic, abdominal and pelvic cavities and their contents
At the completion of necropsy, carcasses were disposed of according to Calvert SOP
Statistics:
Body weights were summarized using descriptive statistics (mean and standard deviation).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Mortality was observed in two female animals at 2000 mg/kg (day 3 and 5).
Clinical signs:
Abnormal gait and stance, hunched posture, decreased activity, decreased body tone, piloerection, black fur around eyes, yellow wet fur of the lower ventral area were observed throughout the study. All animals exhibited necrosis at the application site during the study.
Body weight:
A general decline in body weight was seen in the majority of animals on Day 8; however, by Day 15 body weights were comparable to Day 1.
Gross pathology:
No visible lesions were observed in the surviving animals at terminal necropsy. Dark red intestines were observed in the animal found dead on Day 5 of the study. No lesions were observed in the animal found dead on Day 3 of the study.
Other findings:
No data
Interpretation of results:
GHS criteria not met
Conclusions:
Based upon the results of the acute dermal toxicity study in rats with the substance, the estimated LD50 was considered to be greater than 2000 mg/kg. Therefore, according to the criteria of the CLP Regulation, the substance should not be classified for acute dermal toxicity.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute toxicity: oral

René E Vasquez (2012) assessed the acute oral toxicity of the test substance in Sprague-Dawley rats (up-and-down procedure). The study was carried out based on the guidelines described in: EPA OPPTS 870.1100 "Acute Oral Toxicity" and OECD No. 425, "Acute Oral Toxicity - Up-and-Down procedure". This study is selected as key study.

The first animal was dosed at an initial dose level of 550 mg/kg. Since this animal survived, the second animal received a higher dose (1000 mg/kg). Two additional animals were dosed at 1000 mg/kg, one at 320 mg/kg and one at 2000 mg/kg. The dose for each successive animal was adjusted up or down, depending on the previous result.

Mortality was not observed in any of the animals dosed at 320 or 550 mg/kg of the test article. Two or three animals receiving the test article at 1000 mg/kg were found dead as well as the single animal dosed at 2000 mg/kg.

Clinical observations observed included piloerection at 30 minutes in the single animal at 320 mg/kg and in the surviving animal at 1000 mg/kg. No clinical signs were observed prior to death in the two other animals at 1000 mg/kg. Piloerection, decreased activity, decreased body tone and abnormal gait and stance were observed at 4 hours in the animal treated at 550 mg/kg. At 2000 mg/kg, decreased body tone and abnormal gait were observed at 30 minutes and death was observed at 4 hours.

No biological significant effect was seen on body weights of the surviving animals on Days 8 and 15.

Terminal necropsy revealed no visible lesions in the animals at 320, 550 or the surviving animal at 1000 mg/kg. Necropsy of two animals found dead at 1000 and 2000 mg/kg revealed dark red fluid-filled intestines. The third animal found dead had no visible lesions.

Based on the results of this study, the oral LD50 for the substance in rats was estimated to be 1000 mg/kg.

In addition, in another acute oral toxicity study (class method), F.M. van Otterdijk (2003) observed an LD50 value between 200 and 2000 mg/kg.

Acute toxicity: dermal

Based upon the results of the Acute Dermal Toxicity Study in rats with the substance, the estimated LD50 was considered to be greater than 2000 mg/kg. Mortality was observed in two female animals at 2000 mg/kg. Abnormal gait and stance, hunched posture, decreased activity, decreased body tone, piloerection, black fur around eyes, yellow wet fur of the lower ventral area were observed throughout the study. All animals exhibited necrosis at the application site during the study. Gross necropsy revealed following information: no visible lesions were observed in the surviving animals at terminal necropsy. Dark red intestines were observed in the animal found dead on Day 5 of the study. No lesions were observed in the animal found dead on Day 3 of the study.

Acute toxicity: inhalation

No further acute toxicity studies (inhalation) were performed as the substance is concluded to be corrosive to the skin.

Justification for classification or non-classification

Based on the available acute oral toxicity study, the LD50 is 1000 mg/kg bw/d. The substance should thus be classified as acute oral toxic category 4 (for substances having an LD50 value between 300 and 2000 mg/kg bw); H302 according to CLP Regulation criteria. This conclusion is in line with the RAC opinion proposing harmonised classification and labelling at EU level of the test substance, adopted 9 December 2016 (CLH-O-0000001412-86-132/F).

No classification is warranted for acute toxicity via the dermal route.