Registration Dossier

Administrative data

Description of key information

LD50 oral, rat: ca. 1000 mg/kg (BASF AG, 1997)

LD50 dermal, rat: > 4000 mg/kg (BASF Ag, 1997)

LC50 inhalation (aerosol), rat: 0.371 mg/L (Triazine Task force, 2011)

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: PK 95/222

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: refridgerated
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Biberach
- Age at study initiation: Young adult animals
- Weight at study initiation: 150-300 g
- Fasting period before study: 16 h
- Housing: Single housing in fully air-conditioned rooms.
- Diet: ad libitum, Kliba-Labordiaet (Klingentakmuehle AG, Kaiseraugst, Switzerland)
- Water: ad libitum, tap water
- Acclimation period: for at least 1 week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12 h/12 h

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
The animals were treated by single gavage after having been fasted for 16 hours.
Doses:
500, 1000 and 2000 mg/kg bw
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: Several times on the day of administration, at least once each workday for the individual animal.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology.
Statistics:
LD50 values for male and female rats were calculated with the probit model (Finney, 1971).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
ca. 1 000 mg/kg bw
Based on:
test mat.
Mortality:
4 male and all female animals of the 2000 mg/kg bw dose group, 2 male and 4 female rats of the 1000 mg/kg bw dose group died within 2 days after application.
Clinical signs:
Signs of toxicity noted in the 2000, 1000 and 500 mg/kg bw dose groups comprised impaired or poor general state, dyspnoea, apathy, abdominal or lateral position, staggering, ataxia, atonia, paresis, themor, cyanosis and diarrhoea.
Body weight:
Body weight gain was generally observed in the course of the study.
Gross pathology:
Necropsy findings of the animals that died comprised agonal congestion, erythema, parietal edema and erosion/ulcer in the glandular stomach and discoloration of the mucosa of the forestomach and the glandular stomach.
Histopathological examination of the stomach revealed edema of submucosa in the glandular stomach, margo plicatus and at transition to forestomach, mixed-cell infiltration of submucosa and hyperdemia.
No abnormalities were noted at necropsy of animals sacrificed at the end of the study.

Dose

(mg/kg bw)

Mortality

(Died/Treated)

1 hour

2 hours

3 hours

4 hours

1 day

2 days

14 days

male

female

male

female

male

female

male

female

male

female

male

female

male

female

500

0/5

0/5

0/5

-

-

0/5

0/5

-

0/5

0/5

-

0/5

0/5

0/5

1000

0/5

0/5

0/5

-

-

1/5

0/5

-

2/5

3/5

-

4/5

2/5

4/5

2000

1/5

2/5

2/5

-

-

2/5

3/5

-

4/5

5/5

-

-

4/5

-

Interpretation of results:
Category 4 based on GHS criteria
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
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 001175MCAO

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature; avoid temperatures > 40°C
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Netherlands, Horst
- Age at study initiation: Young adult males and females (about 8 - 12 weeks)
- Weight at study initiation: Animals of comparable weight (180 - 300 g) (± 20% of the mean weight)
- Housing: up to 5 animals per cage
- Diet: Kliba-Labordiät (Maus/Ratte Haltung "GLP"), Provimi Kliba SA, Kaiseraugst, Basel, Switzerland, ad libitum
- Water: Tap water ad Iibitum
- Acclimation period: Acclimatization period at least 5 days before the beginning of the experimental phase; during the acclimatization period, the
animals are accustomed to the environmental conditions of the study and to the diet.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose/head only
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetric measurements
Duration of exposure:
4 h
Concentrations:
0.25, 0.5 and 1 mg/L (target concentrations)
0.257, 0.485 and 1.068 mg/L (analytical concentrations)
No. of animals per sex per dose:
5
Control animals:
other: control animals for examination of local effect of the test item in satellite group
Details on study design:
- Duration of observation period following administration: 14 days for main groups, 1 day for satellite group including control group
- Frequency of observations and weighing: A check for moribund and dead animals was made twice each workday and once daily at weekends and on public holidays. Detailed clinical observations were recorded for each animal separately several times during exposure and at least once daily on the pre-exposure day and during the observation period. Individual body weight was determined once during the acclimatization period, at the start of exposure period (day 0) and on days 1, 3 and 7, and weekly thereafter or before the sacrifice of the animals at the end of the observation period. The body weights of1 animals that died or were sacrificed in a moribund state were determined as they were discovered from study day 1 onward.
- Necropsy of survivors performed: yes
- Other examinations performed: concentration of monoethanol amine (MEA) and formaldehyde (FA) in the test atmosphere; organ weight of lung and histological examination of respiratory tract in satellite group animals
- Satellite group: To examine the local effect of the test item, a satellite group of 5 male animals was exposed to the test substance together with the animals of the first exposure group (target concentration 1 mg/L). These animals were sacrificed on study day 1 and subjected to histological examination. Two male animals were exposed to air and were sacrificed with the satellite group animals. They served as control for the histological examinations.
Statistics:
The LC50 was calculated by Probit analysis (FINNEY, D.J. (1971): "Probit Analysis" Cambridge University Press) by means of a computer program. The calculation of the particle size distribution was carried out in the inhalation laboratory on the basis of mathematical methods for evaluating particle measurements (DIN 66141: Darstellung von Korngrößenverteilungen, DIN 66161: Partikelgrößenanalyse (Beuth-Vertrieb GmbH, Berlin 30, FRG und Köln 1, FRG)).
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
0.371 mg/L air (analytical)
Based on:
test mat.
95% CL:
0.229 - 0.529
Exp. duration:
4 h
Sex:
male
Dose descriptor:
LC50
Effect level:
0.4 mg/L air (analytical)
Based on:
test mat.
95% CL:
0.167 - 0.943
Exp. duration:
4 h
Sex:
female
Dose descriptor:
LC50
Effect level:
0.338 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
Two of the five female animals and one of the five male animals died at 0.257 mg/L. Three of the five female animals and three of the five male animals died at 0.485 mg/L. At 1.068 mg/L all of the male and female animals died or were sacrificed in a moribund state. Lethality was observed either during exposure or shortly after exposure (day 0 to 2).
Clinical signs:
other: Clinical signs of toxicity in animals exposed to 0.257 mg/L comprised accelerated respiration, labored respiration, abdominal respiration, gasping, respiration sounds, colorless discharge of the nose, red encrusted nose, distended abdomen, piloerection, y
Body weight:
Test group 1 (0.257 mg/L): The mean body weights of the surviving animals decreased during the first post exposure observation week but increased during the second week.
Test group 2 (0.485 mg/L): The mean body weights of the surviving animals decreased during the first post exposure observation week but increased during the second week.
Test group 3 (1.068 mg/L): The mean body weights of the surviving animals decreased until the death of the animals.
Gross pathology:
Test group 1 (0.257 mg/L): No gross pathological abnormalities were detected in the female animal that died on study day 0. In the two animals that died on study day 2 (1 male + 1 female), dilation with gaseous content of the stomach and small intestine were found. No gross pathological abnormalities were detected during necropsy in the surviving animals at the termination of the post exposure observation period.
Test group 2 (0.485 mg/L): Gross pathological abnormalities were noted neither in animals died on day 0/day 1, nor in surviving animals at the termination of the post exposure observation period.
Test group 3 (1.068 mg/L): During gross necropsy dilation with gaseous content of the stomach and/or small intestine were observed in individual animals. In other animals no gross pathological abnormalities were noted.
Other findings:
ANALYTICAL INVESTIGATIONS:
Concentration measurements: The following measured concentrations were tested: 0.257, 0.485 and 1.068 mg/L (analytical concentration). Identity of the test substance was confirmed by off-line infra-red spectroscopy. Moreover, very small fractions of formaldehyde (mean concentrations 0.48 to 0.73 ppm) were detected in the test atmospheres. The mean concentration of monoethanol amine in the test atmospheres was around 0.6 mg/m³, corresponding to 0.0006 mg/L air.
Particle size analysis: Cascade impactor measurements resulted in particle size distributions with mass median aerodynamic diameters (MMADs) between 1.6 and 1.9 μm, which are well within the respirable range.

SATELLITE GROUP:
Satellite animals and its concurrent controls were designated for examination of the respiratory tract by light microscopy on study day 1. Due to the high toxicity however, some of these animals were found dead on study day 1, others were sacrificed in moribund state. Nevertheless, histopathological examination of the respiratory tract revealed up to severe findings in the upper and lower part. Especially the upper part of the respiratory tract (nasal cavity and larynx) was severely affected. The findings were characterized by multifocal to coalescing up to diffuse necrosis of the different epithelia in the nasal cavity, with accompanying inflammation in the moribund sacrificed animals. Also the laryngeal epithelium showed a diffuse necrosis in all levels in the treated animals with inflammation in the surrounding tissue in some animals. The trachea and the lung revealed also slight to minimal treatment-related findings (hemorrhage, edema, emphysema, necrosis) in some animals. In addition, the respiratory tract of one representative female animal was also examined by light microscopy. Similar lesions as those in male animals were observed.
Interpretation of results:
Category 2 based on GHS criteria
Conclusions:
HHT caused concentration-related severe clinical symptoms and mortality after 4-hour head/nose aerosol exposure. Histopathology performed at the high concentration revealed severe lesion at the entire respiratory tract. The death of several test animals is probably a direct consequence of severe diffuse necrosis of laryngeal and nasal epithelium. Under the conditions of this study the LC50 for male and female rats after liquid aerosol
inhalation exposure of HHT was determined to be 0.371 mg/L (analytical concentration).
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Value:
371 mg/m³

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Principles of method if other than guideline:
A single dermal dose of the undiluted test substance was applied to one group of 10 Wistar rats (5 males and 5 females) at a dose level of 4000 mg/kg bw (3.43 mL/kg). The test material was applied to the clipped epidermis (dorsal and dorsolateral parts of the trunk) of each animal and was covered by a semiocclusive dressing for 24 hours. Post-observation period: 14 days.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: PK 95/222

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: refridgerated
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Biberach
- Age at study initiation: Young adult animals
- Weight at study initiation: 200-300 g
- Fasting period before study: > 15 h
- Housing: Single housing in fully air-conditioned rooms.
- Diet (ad libitum): Kliba-Labordiaet 343 (Klingentakmuehle AG, Kaiseraugst, Switzerland)
- Water (ad libitum): tap water
- Acclimation period: for at least 1 week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12 h/12 h
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: about 50 cm²
- Type of wrap: The bandage consisted of four layers absorbent Gauze and fixomull stretch (adhesive fleece).


REMOVAL OF TEST SUBSTANCE
- Washing: with warm water
- Time after start of exposure: 24 h


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 3.43 mL/kg
- Concentration: undiluted


Duration of exposure:
24 h
Doses:
4000 mg/kg bw
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: Individual readings 30 - 60 min. after removal of the semiocclusive dressing (day 1), at least once each workday.
- Necropsy of survivors performed: yes.
- Other examinations performed: clinical signs, body weight, scoring of skin findings.
Statistics:
Statistical analysis: Binomial test (Snedecor and Cochran, 1989).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 4 000 mg/kg bw
Mortality:
No mortality observed.
Clinical signs:
Systemic signs of toxicity were not noted.
Body weight:
Body weight gain was generally observed in the course of the study.
Gross pathology:
No pathologic findings noted. No abnormalities were noted at necropsy of animals sacrified at the end of the study.
Other findings:
The following local effects were observed 1 day or 7 days after application: very slight, well-defined or moderate erythema, very slight to slight edema, scaling and superficial scabbing.
Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
4 000 mg/kg bw

Additional information

Oral:

The key study was performed according to OECD TG 401 in compliance with GLP (BASF AG, 1997). To each group of 10 fasted Wistar rats (5 males and 5 females) a single oral dose of the test substance at dose levels of 2000, 1000 and 500 mg/kg bw was given, followed by a post-observation period of 14 days. An oral LD50 of approx. 1000 mg/kg bw was determined. Signs of toxicity noted in the 2000, 1000 and 500 mg/kg bw dose groups comprised impaired or poor general state, dyspnoea, apathy, abdominal or lateral position, staggering, ataxia, atonia, paresis, themor, cyanosis and diarrhoea. Necropsy findings of the animals that died comprised agonal congestion, erythema, parietal edema and erosion/ulcer in the glandular stomach and discoloration of the mucosa of the forestomach and the glandular stomach. Histopathological examination of the stomach revealed edema of submucosa in the glandular stomach, margo plicatus and at transition to forestomach, mixed-cell infiltration of submucosa and hyperdemia. No abnormalities were noted at necropsy of animals sacrificed at the end of the study.

Comparable oral LD50 values in rats (between 763 - 1850 mg/kg) have been reported in other company data (BASF, 1973) and in public literature (Siglin, 1992; Passman 1995). In contrast, in an acute oral toxicity study according to OECD guideline 423 in compliance with GLP no mortality occured in female Sprague Dawley rats after adminstration of 2000 mg/kg bw per gavage (LACSA, 2015).

In this study, the LD50 of the test item SYNTAN OXB was greater than 2000 mg/kg body weight by oral route in the rat.

 

Dermal:

The key study was performed according to OECD TG 402 in compliance with GLP (BASF AG, 1997). A single dermal dose of the undiluted test substance was applied to one group of 10 Wistar rats (5 males and 5 females) at a dose level of 4000 mg/kg bw (3.43 mL/kg). The test material was applied to the clipped epidermis (dorsal and dorsolateral parts of the trunk) of each animal and was covered by a semiocclusive dressing for 24 hours. A post-observation period of 14 days followed. No mortality was observed (LD50 > 4000 mg/kg bw). No signs of systemic toxicity, no pathologic findings and no abnormalities at necropsy were noted.

This result is supported by comparable data from other companies (GLP guideline study in Sprague Dawley rats; L.A.C.S.A, 2015) and public literature (Siglin, 1992; Passman 1995).

 

Inhalation:

An acute inhalation toxicity study in rats with 4-hour head/nose liquid aerosol exposure was performed according to OECD guideline 403 in compliance with GLP (Triazine Task Force, 2011). In this study, HHT caused concentration-related severe clinical symptoms and mortality after 4-hour head/nose aerosol exposure at concentrations of 0.25, 0.5 and 1 mg/L. Histopathology performed at the high concentration revealed severe lesion at the entire respiratory tract. The death of several test animals was probably caused by a direct consequence of severe diffuse necrosis of laryngeal and nasal epithelium. Under the conditions of this study the LC50 for male and female rats after liquid aerosol inhalation exposure of HHT was determined to be 0.371 mg/L (analytical concentration).

Supportingly, the respiratory effects of two synthetic metalworking fluids and their components were evaluated in two inhalation studies in mice (Krystofiak and Schaper, 1996; Detwiler-Okabayashi and Schape, 1996). HHT was identified as an irritating component in both fluids. Male Swiss-Webster mice exposed to HHT at 112 - 351 mg/m³ exhibited signs of both sensory and pulmonary irritation during the 3-hour exposure. In both studies, HHT resulted in delayed deaths of mice at 24 to 72 hours post exposure. The concentrations of HHT capable of evoking a 50 percent decrease in mean respiratory frequency based on pulmonary irritation (RD50) were calculated as 137 mg/m³ in the study by Krystofiak and Schaper (1996) and as 190 mg/m³ in the study by Detwiler-Okabayashi and Schaper (1996).

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The oral LD50 was determined to be 1000 mg/kg bw. The inhalative LC50 (aerosol) is 0.371 mg/L. The dermal LD50 exceeds the limit of 4000 mg/kg bw. As a result the substance is considered to be classified as cat. 4 (H302) and cat. 2 (H330) under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation (EU) No 2016/ 1179.