Registration Dossier

Administrative data

Description of key information

Oral (OECD 420): 2000 mg/kg bw < LD50 (female rat) < 5000 mg/kg bw. 
Dermal: No data available
Inhalation (OECD 436): LC50 > 4700 mg/m³ air

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:
13 October 2011 to 08 November 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
(2001)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
(2002)
Deviations:
no
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Rat, RccHan: WIST(SPF)
- Source: Harlan Laboratories B.V., Kreuzelweg 53, 5961 NM Horst, The Netherlands
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 171.9 g–197.8 g
- Fasting period before study: overnight fasting period prior to treatment and approximately 3 hours post dose
- Housing: In groups of up to five in Makrolon type-4 cages with wire mesh tops and standard softwood bedding (‘Lignocel’ J. Rettenmaier & Söhne GmbH&CoKG, 73494 Rosenberg, Germany, imported by Provimi Kliba AG, 4303 Kaiseraugst, Switzerland). Paper enrichment, batch no. 75, (Enviro-dri from Lillico Biotechnology, Surrey, UK) was included.
- Diet: Pelleted Harlan Teklad 2914C rodent maintenance diet (batch no. 46/11, Provimi Kliba AG, 4303 Kaiseraugst, Switzerland), ad libitum
- Water: Community tap water from Itingen, ad libitum
- Acclimation period: 5-12 days under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 30-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light):12/12

IN-LIFE DATES: From: 18 October 2011 to: 08 November 2011
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 1.94 ml/kg bw at the 2000 mg/ kg bw dose level
Doses:
sighting study: 300 and 2000 mg/kg bw
main study : 2000 mg/kg bw
No. of animals per sex per dose:
sighting study: 1 female at 300 mg/kg and 1 female at 2000 mg/kg
main study: 4 females at 2000 mg/kg
Control animals:
other: not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2-15
- Frequency of weighing: day 1 (prior to administration) and on test days 8 and 15
- Necropsy of survivors performed: yes
Statistics:
No statistical analysis was performed.
Preliminary study:
The sighting study started with a single female at a dose of 300 mg/kg body weight. As no clinical signs were observed at the initial dose, the next higher dose level of 2000 mg/kg investigated, using a single female. As no mortality occurred at 2000 mg/kg, the sighting study was complete and the main study was conducted at 2000 mg/kg.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal treated at 2000 mg/kg was found dead on test day 2. All other animals survived the scheduled treatment and observation periods.
Clinical signs:
In animal no. 1 treated at 300 mg/kg, decreased activity, hunched posture and ruffled fur were noted on test day 1. Thereafter, the animal was free of clinical signs up to the end of the observation period. In the animals treated at 2000 mg/kg, dragging of limbs, decreased activity, hunched posture, ruffled fur and / or swaying gait were noted on test day 1 and persisted up to test day 3 at the latest. Thereafter, no clinical signs were noted up to test day 15, the end of the observation period.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
In animal no. 6 which died spontaneously, distended stomach was recorded at necropsy. No abnormal macroscopic findings were recorded in the remaining animals at scheduled necropsy.
Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulations (EC) No 1272/2008
Conclusions:
The test item was tested for acute oral toxicity according to the OECD TG 420, and in compliance with GLP. In the main study one animal treated at 2000 mg/kg was found dead on test day 2. All other animals survived the scheduled treatment and observation periods. The clinical signs recorded at this dose level were dragging of limbs, decreased activity, hunched posture, ruffled fur and/or swaying gait on test day 1 and persisted up to test day 3 at the latest. Based on Annex 3 and 4 of the OECD TG 420, the test item is classified as GHS category 5 corresponding to 2000 mg/kg body weight < LD50 (female rat) < 5000 mg/kgbw. Classification for acute oral toxicty according to EC/1272/2008 is not warranted.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The study was conducted according to an appropriate OECD guideline, and in compliance with GLP.

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
Study period:
09 December 2011 to 23 December 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Version / remarks:
(2009)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Swiss Confideration, Swiss Federal Office of Public Health, Consumer protection directorate, Notification authority for chemicals, Bern, Switzerland
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: RccHan:WIST(SPF)
- Source: Harlan Laboratories B.V. (Kreuzelweg 53, 5961 NM Horst, Netherlands)
- Age at study initiation: 9 weeks
- Weight at study initiation: 278.0-282.6 g (males) and 181.8-193.5 g (females) The weight variation did not exceed ±4% of the mean weight of the corresponding sex.
- Fasting period before study: none
- Housing: in groups of 3 of the same sex in Makrolon® type-IV cages with wire mesh tops and standard softwood bedding ("Lignocel" J. Rettenmaier & Söhne GmbH & Co KG, 73494 Rosenberg, Germany, imported by Provimi Kliba AG, 4303 Kaiseraugst, Switzerland) including paper enrichment (Enviro-dri from Lillico Biotechnology, Surrey, UK)
- Diet: pelleted standard Harlan Teklad 2914C rat maintenance diet (Provimi Kliba AG, 4303 Kaiseraugst, Switzerland) batch no. 44/11, ad libitum except during the period when the animals were restrained in exposure tubes
- Water: Community tap water from Füllinsdorf in water bottles, ad libitum except during the period when they were restrained in exposure tubes
- Acclimation period: 9 days under optimal hygienic laboratory conditions; the animals were accustomed to the restraining tubes for 30 minutes on the day of exposure

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12 (A radio program was played during most of the light period.)

IN-LIFE DATES: from 09 December 2011 to 23 December 2011
Route of administration:
inhalation: vapour
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Inhalation exposure was performed using a flow-past, nose-only exposure system. The exposure system ensured a uniform distribution and provided a constant flow of test material to each exposure tube.
- Exposure chamber volume:
- Method of holding animals in test chamber: The animals were confined separately in restraint tubes which were positioned radially around the exposure chamber.
- Rate of air: The flow of air at each tube was 1.0 L/min, which is sufficient to minimize re-breathing of the test atmosphere as it is more than twice the respiratory minute volume of rats.
- System of generating vapour: The test atmosphere was generated using a glass nebulizer with a metal injector connected to a syringe pump. The temperature of the glass nebulizer was kept at 70°C to maximize the vaporization of the test item. A particle filter was placed before the exposure chamber to ensure that any aerosol droplets were retained and that the animals were exposed only to the vapour phase.
- Temperature, humidity, oxygen in air chamber: 22.3-22.4 °C, 2.9-3.0% relative humidity, 20.2% (v/v) oxygen concentration

TEST ATMOSPHERE
- Brief description of analytical method used: The nominal concentration was determined during exposure by weighing the nebulizer and appropriate adjacent pipe work, before and after exposure to determine the quantity of test item used for test atmosphere generation. The weight used was then divided by the total airflow volume to give the nominal concentration.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: technically highest achievable concentration as vapour
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Test atmosphere samples were collected 4 times during exposure in a solvent trap with 100 ml acetonitrile at 20±5 °C. The duration of sampling was 10-12 min. The samples were stored at -20±5 °C until analysis using a GC method.
Duration of exposure:
4 h
Concentrations:
- Target concentration: approx. 4.5 mg/l air
- Nominal vapour concentration: 7.0 mg/ml air
- Analytical concentration: 4.7 mg/ml air
No. of animals per sex per dose:
3
Control animals:
other: not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations for viability/mortality: Observations for viability were recorded once before exposure on the day of exposure (test day 1), three times during exposure, immediately and 1 h after exposure on test day 1 and twice daily during the observation period.
- Frequency of observations for clinical signs: Each animal was examined three times during exposure, immediately and 1 h after exposure on test day 1 and once daily during the observation period.
- Frequency of weighing: The body weight of each animal was recorded on test days 1 (before exposure), 2, 4, 8 and 15 (before necropsy).
- Necropsy of survivors performed: yes. Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered 4% formaldehyde solution (head with nasopharyngeal tissues, larynx, lungs: instilled via trachea with formalin at approximately 30 cm H2O pressure, trachea). All collected organ and tissue samples were retained but neither processed nor examined.
- Other examinations performed: clinical signs, body weight
Statistics:
No statistical analysis was performed as only one group was allocated to the study.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4 700 mg/m³ air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: No mortality occured. The test concentration of 4700 mg/m³ air was the highest vapour concentration achievable.
Mortality:
All animals survived the scheduled observation period.
Clinical signs:
During exposure, salivation and reddish nasal secretion were observed in all three males and in one or two females, respectively. After the end of treatment, ruffled fur was noted for all animals. In addition, decreased activity and hunched posture were recorded for two males and in two or all three females, respectively. There were no clinical signs recorded from test day 2 until the end of the observation period.
Body weight:
From test day 1 to test day 2, slight body weight loss was noted in all animals. Thereafter normal body weight development was recorded.
Gross pathology:
No macroscopic findings were present at necropsy.

TEST ATMOSPHERE CONDITIONS

Temperature, relative humidity and oxygen concentration during exposure were considered to be

satisfactory for this type of study.

Tab. 1: Data on temperature, relative humidity and oxygen concentration

Recording Time

[hours/min]

 

O2Concentration

[Vol %]

 

Temperature

[°C]

 

Relative Humidity

[% RH]

07:00

20.2

22.2

3.0

07:30

20.2

22.3

3.0

08:00

20.2

22.3

3.0

08:30

20.2

22.3

2.9

09:00

20.2

22.4

2.9

09:30

20.2

22.4

2.9

10:00

20.2

22.4

2.9

10:30

20.2

22.4

2.9

11:00

20.2

22.4

2.9

Mean

20.2

22.3

2.9

SD

0.0

0.0

0.0

N

9

9

9

CHEMICAL DETERMINATION OF VAPOR CONCENTRATION

Tab. 2: Details on chemically determined vapor concentrations

Sampling Time

[hours/min]

 

Sampling Volume

[l]

 

Amount of Test Item in

the Solvent Trap

[mg]

 

Chemical Vapor

Concentration

[mg/l air]

07:26-07:36

9.8

46.2

4.7

08:34-08:46

11.8

57.0

4.8

09:17-09:27

9.8

45.4

4.6

10:15-10:27

11.8

55.4

4.7

          Mean

4.7

          SD

0.1

          N

4

Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulations (EC) No 1272/2008
Conclusions:
The test item was tested for acute inhalation toxicity according to the OECD TG 436, and in compliance with GLP. Treatment with the test item at a concentration of 4700 mg/m³ air for 4 h resulted in clinical signs (salivation, reddish nasal secretion, ruffled fur, decreased activity and hunched posture) and body weight loss. This observation exceeded the marginal body weight loss or stagnation of the body weight gain which is usually observed in acute inhalation studies. However, the restraining of the animals in the tubes during the nose-only exposure procedure may have added to the observed effect. In conclusion, the LC50 was > 4700 mg/m³ air, which was the technical limit concentration for a vapour. Hence, classification for actute inhalation toxicity according to EC/1272/2008 is not warranted.
Executive summary:

A group of three male and three female albino rats [RccHanTM:WIST(SPF)] was exposed by nose-only, flow-past vapor inhalation for four hours to the test item at a chemically determined mean concentration of 4.7 mg/l air. All animals were observed for clinical signs and mortality during the inhalation exposure and the subsequent 14-day observation period. Body weights were recorded prior to exposure on test day 1, and during the observation period on test days 2, 4, 8 and 15 before necropsy. On test day 15 all animals were sacrificed and necropsied. The vapor concentration, temperature, relative humidity, oxygen content and airflow rate measured during the exposure were considered to be satisfactory for a study of this type. In addition, as a vapor the test item was considered to be respirable to rats. All animals survived the scheduled observation period. Most animals showed salivation and reddish nasal secretion during exposure and ruffled fur, decreased activity and hunched posture thereafter. There were no clinical signs recorded from test day 2 onwards. From test day 1 to test day 2, slight body weight loss was noted in all animals. Thereafter normal body weight development was recorded. No macroscopic findings were present at necropsy. In conclusion, the LC50 for 4-hour exposure of (chloromethyl)triethoxysilane obtained in this study was greater than 4.7 mg/l air (chemically determined mean vapor concentration). This concentration was at the technical limit for a vapor and considered to be close to the saturation concentration. There was no indication of relevant sex-related differences in toxicity of the test item.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The study was conducted according to an appropriate OECD test guideline, and in compliance with GLP.

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Acute toxicity: oral

In the available key study (Harlan, 2012a) the test item was investigated for acute oral toxicity according to the OECD TG 420, and in compliance with GLP. To determine the dose level for the main study, two sighting studies were conducted with one female Wistar rat each. The two animals were treated sequentially with the undiluted test item at the dose levels of 300 or 2000 mg/kg bw, respectively, by single oral gavage administration. All animals were examined for mortality and clinical signs before treatment, within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2-15. Body weights were recorded on test day 1 (prior to administration) and on test days 8 and 15. All animals were necropsied and macroscopically examined.

Since no mortality occurred during the sighting study up to 2000 mg/kg, additional four females were treated at this dose level in the main study. One animal treated at 2000 mg/kg was found dead on day 2 of the main study. All other animals survived the scheduled treatment and observation periods. In the animal treated at 300 mg/kg during sighting study, decreased activity, hunched posture and ruffled fur were noted on test day 1. Thereafter, the animal was free of clinical signs up to the end of the observation period. In the animals treated at 2000 mg/kg, dragging of limbs, decreased activity, hunched posture, ruffled fur and/or swaying gait were noted on test day 1 and persisted up to test day 3 at the latest. Thereafter, no clinical signs were noted up to study termination on day 15. The body weight of the animals was within the range commonly recorded for this strain and age. In the animal that died spontaneously, distended stomach was recorded at necropsy. No abnormal macroscopic findings were recorded in the remaining animals at scheduled necropsy. Based on this data, classification for acute oral toxicity accroding to 67/584/EEC and EC/1272/2008 is not warranted.

Acute toxicity: dermal

No data available.

Acute toxicity: inhalation

In the available key study (Harlan, 2012b) the test item was tested for acute inhalation toxicity according to the OECD TG 436, and in compliance with GLP. A group of three male and three female Wistar rats was exposed by nose-only, flow-past inhalation for four hours to the test item at a chemically determined mean concentration of 4700 mg/m³ air. All animals were observed for clinical signs and mortality during the inhalation exposure and the subsequent 14-day observation period. Body weights were recorded prior to exposure on test day 1, and during the observation period on test days 2, 4, 8, and 15 before necropsy. On test day 15 all animals were sacrificed and necropsied. All animals survived the scheduled observation period. Most animals showed salivation and reddish nasal secretion during exposure and ruffled fur, decreased activity and hunched posture thereafter. There were no clinical signs recorded from test day 2 onwards. From test day 1 to test day 2, slight body weight loss was noted in all animals. Thereafter normal body weight development was recorded. No macroscopic findings were present at necropsy. In conclusion, the LC50 was greater than 4700 mg/m³ air, which was at the technical limit for a vapour. There was no indication of relevant sex-related differences in toxicity of the test item.


Justification for classification or non-classification

The available data are reliable and suitable for classification. Based on these data, classification for acute toxicity according to 67/584/EEC and EC/1272/2008 is not warranted.