Registration Dossier

Administrative data

Description of key information

aktue oral toxicity: LD50 > 2000 mg/kg bw
acute inhalation toxicity: LC50 > 5142 mg/m³

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:
other: GLP guideline study
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)
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
SPF-bred Wistar rats of the strain Clr:(WI)BR
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: about 10 weeks
- Fasting period before treatment: overnight (maximal 20 h)
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Doses:
2000 mg/kg
No. of animals per sex per dose:
3
Control animals:
no
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
no deaths
Clinical signs:
Clinical signs as lethargy, hunched posture, laboured respiration, rales, piloerection, uncoordinated movements, and/or chromodacryorrhea were noted among the animals between days 1 and 7.
Body weight:
no effects on weight gain
Gross pathology:
no abnormal findings
Other findings:
none

Dose (mg/kg bw) Toxicological results  Onset and duration of signs  Onset of mortality
2000 0 / 3 / 3 --- --- 
2000 0 / 3 / 3 --- --- 

Toxicological results:

number of dead animals / number of animals with clinical signs after treatment / number of animals treated

Executive summary:

The acute oral toxicity of the test item was evaluated in a GLP-compliant study on male and female Wistar rats according to OECD TG 423. The oral dose of 2000 mg/kg bw was tolerated without mortalities, effects on weight gain or gross pathological findings. Clinical signs as lethargy, laboured respiration, piloerection, and/or chromodacryorrhea were noted among the animals between days 1 and 7. The LD50 was determined with > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 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:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Guideline:
other: test procedures were adapted so as to comply also with the EU Directive 92/69/EEC and OECD Guidance Document No. 39 (2009)
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: HsdRCCHan: Wist (SPF bred)
- Source: Harlan-Nederland (NL), AD Horst
- Age at study initiation: approximately 2 months
- Weight at study initiation: At the study start the variation of individual weights did not exceed ± 10 per cent of the mean for each sex
- Housing: singly in conventional Makrolon® Type IIIH cages
- Diet and water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 40 - 60 %
- Air changes (per hr): approximately 10
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Mode of exposure: Animals were exposed to the aerosolized test article in restrainers made of Plexiglas. The design of the directed-flow inhalation chamber minimizes rebreathing of the exhaled test atmosphere.
- Generation of atmosphere and exposure techniques: Test atmosphere was generated using a WRIGHT DUST FEEDER system (BG IInc.,Waltham, MA 02154, USA). For dry powder dispersion, conditioned compressed dry air (28 liters/min; generic dispersion pressure: 120 kPa) was used. The principle
performance of the WRIGHT DUST FEEDER dust generating system can be described as follows: the test substance was metered in a reservoir and then was compressed to a pellet using approximately 1 metric ton by a carva laboratory press (F. S. Carver Inc., Wabash, IN 46992, USA). From this pellet defined amounts of test substance were scraped off and entrained into the main air flow. The airborne powder was then conveyed into the inner cylinder of the inhalation chamber.
- Inhalation chamber equilibrium concentration: The test atmosphere generation conditions provide an adequate number of air exchanges per hour [28 L/min x 60 min/(3.8 L) = 422, continuous generation of test atmosphere]. Based on OECD GD39 the equilibrium concentration (t95) can be calculated after McFarland, 1976. Thus, under the test conditions used chamber equilibrium is attained in less than one minute of exposure. At each exposure port a minimal air flow rate of 0.75 L/min was provided. The test atmosphere can by no means be diluted by bias-air-flows.
- Conditioning the compressed air: Compressed air was supplied by Boge compressors and was conditioned (i.e. freed from water, dust, and oil) automatically by a VIA compressed air dryer. Adequate control devices were employed to control supply pressure.
- Exhaust air treatment: The exhaust air was purified via filter systems.
- Temperature and humidity measurements were performed by a computerized Data Acquisition and Control System using HC-S3 sensors (Rotronic). The position of the probe was at the exposure location of rats. Temperature and humidity data are integrated for 30-seconds and displayed accordingly. The humidity sensors are calibrated using saturated salt solutions according to Greenspan (1977) and Pauluhn (1994) in a two-point calibration at 33 % (MgCI2) and at 75 % (NaCI) relative humidity. The calibration of the temperature sensors is also checked at two temperatures
using reference thermometers.

TEST ATMOSPHERE
- Nominal concentration: The nominal concentration was not calculated from the ratio of the total quantity of test article consumed and the total throughput of air through the inhalation chamber as this would have required a dismantling of the dust generator.
- Samples taken from breathing zone: yes
- Gravimetric concentation: gravimetric analysis of filter samples (filter: Glass-Fibre-Filter, Sartorius, Göttingen, Germany; digital balance).
- Particle size distribution: The particle-size distribution was analyzed using an ANDERSON critical orifice cascade impactor. An adhesive stage coating (silicone spray) was omitted due to the adhesive properties of the test article. Gravimetric analyses of filters used a digital balance.
- MMAD (Mass median aerodynamic diameter): 3.5 µm; GSD (Geometric st. dev.): 2.4
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
5142 mg/m³ (actual concentration as dry powder aerosol)
No. of animals per sex per dose:
5
Control animals:
other: Comparisons with an appropriate historical air control group were performed. This control was exposed to an atmosphere using essentially similar exposure conditions as were used for the test substance (15 L air/min; conditioned air; 4 h exposure)
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Body weights were measured before exposure, on days 1, 3 and 7, and weekly thereafter. The appearance and behavior of each rat were examined carefully several times on the day of exposure and at least once daily thereafter. Weekend assessments were
made once a day (morning). Assessments from restraining tubes were made only if unequivocal signs occurred (e.g. spasms, abnormal movements, and severe respiratory signs).
- Necropsy of survivors performed: yes
- Other examinations performed: The rectaI temperatures were measured shortly after cessation of exposure using a digital thermometer with a rectal probe for rats.
Statistics:
- Body weights: Means and single standard deviations of body weights are calculated. Mean body weights are also depicted graphically as a function of time. Since in acute studies individual group means may differ prior to commencement of the first exposure, the body weight gain was statistically evaluated for each group. For these evaluations a one-way ANOVA (vide infra) is used.
- Physiological data: Data of rectal temperature measurements are statistically evaluated using the ANOVA procedure (vide infra).
- Calculation of the LCso: If calculation of a median lethal concentration (LCSO) is possible, it is performed by computer (PC) according to the method of Rosiello et al. (1977) as modified by Pauluhn (1983). This method is based on the maximumlikeli hood method of Bliss (1938). If only 2 pairs of values with greater than 0 % lethality and less than 100 % are available then the first linear approximation is based on these values and a l-homogeneity test is not performed. In this case the interpolated concentration at SO% lethality is designated the approximate LCSO. Additionally, the moving average interpolation according to Schaper et al. (1994) is used for calculation, if applicable.
- Analysis of variance (A NO VA): This parametric method checks for normal distribution of data by comparing the median and mean. The groups are compared at a confidence level of (1-a) = 9S % (p = O.OS). The test for the between-group homogeneity of the variance employed Box's test if more than 2 study groups were compared with each other. If the above F-test shows that the intra-group variability is greater than the inter-group variability, this is shown in the Appendix as "no statistical difference between the groups". If a difference is found then a pairwise post-hoc comparison is conducted (1- and 2-sided) using the Games and Howell modification of the Tukey-Kramer significance test.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
5 142 mg/m³ air
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: clinical signs: irregular and laboured breathing
Sex:
female
Dose descriptor:
LC50
Effect level:
> 6 597 mg/m³ air
Based on:
act. ingr.
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
other: NOAEL
Effect level:
< 1 554 mg/m³ air
Based on:
act. ingr.
Exp. duration:
4 h
Mortality:
Mortality did not occur up to the maximum technically attainable level.
Clinical signs:
other: Irregular and labored breathing pattern, bradypnea, dyspnea, breathing sounds, motility reduced, atony, high-legged gait, piloerection, and cyanosis.
Body weight:
Comparisons between the control and the exposure groups revealed transient changes (decreases) in body weights.
Gross pathology:
Animals sacrificed at the end of the observation period: The macroscopic findings of extrapulmonary organs were essentially indistinguishable between the test article exposure and control group. In the test group local discolorations of the lung and enlarged lung-associated lymph nodes were observed.
Other findings:
A battery of reflex measurements was made on the first post-exposure day. Differences between the rats of the control and the trated group were not observed.
Statistical comparisons between the control and the exposure groups revealed significant changes in body temperatures indicative of hypothermia.
Executive summary:
A study on the acute inhalation toxicity on rats has been conducted in accordance with OECD TG 403. One group of Wistar rats (5 males and 5 female) were nose only exposed (4 h) to the solid aerosol of the test article in the maximum technically achievable concentration of 5142 mg/m³. The aerosol was generated so that it was respirable to rats (average MMAD of 3.5 µm / GSD 2.4). Mortality did not occur. Clinical observation showed irregular and labored breathing patterns as the lead pathodiagnostic effect. Based on the results the LC50 was determined with 5142 mg/m³ for males and females. In summary, the test substance (solid aerosol) proved to have a low acute inhalation toxicity in rats.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
Value:
5 142 mg/m³

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

An acute oral toxicity study according to OECD TG 423 was conducted with a limit dose of 2000 mg/kg test item administered to 3 male and 3 female rats. No animal died in the course of the study. As clinical signs e.g. lethargy, hunched posture and laboured respiration were described. Body weight development was not affected. The animals sacrificed at the end of study showed no noticeable gross pathological findings. The LD50 was estimated with > 2000 mg/kg bw.

A study on the acute inhalation toxicity on rats has been conducted in accordance with OECD TG 403. One group of Wistar rats (5 males and 5 female) were nose only exposed (4 h) to the dry powder aerosol of the test article in the maximum technically achievable concentration of 5142 mg/m³. The aerosol was generated so that it was respirable to rats (average MMAD of 3.5 µm / GSD 2.4). Mortality did not occur. Clinical observation showed irregular and laboured breathing patterns as the lead pathodiagnostic effect. Based on the results the LC50 was determined with > 5142 mg/m³ for males and females. In summary, the test substance (solid aerosol) proved to have a low acute inhalation toxicity in rats.

No data on acute dermal toxicity are available for the substance. According to Commission Regulation (EU) 2016/863 of May 2016 acute toxicity testing by the dermal route (Annex VII, point 8.5.3., column 2) ‘does not need to be conducted if the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure'. The registered substance conforms with the requirements given above. Therefore, it can be concluded for acute dermal toxicity that the available information is conclusive for non-classification.

Justification for classification or non-classification

Based on the available relevant and reliable data, the test substance does not need to be classified and labelled according to the CLP Regulation (EC) No 1272/2008 with respect to acute oral and inhalation toxicity.

No study is available for acute dermal toxicity. According to Commission Regulation (EU) 2016/863 of May 2016 acute toxicity testing by the dermal route (Annex VII, point 8.5.3., column 2) ‘does not need to be conducted if the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure'. The registered substance conforms with the requirements given above. Therefore, it can be concluded for acute dermal toxicity that the available information is conclusive for non-classification.