Registration Dossier

Administrative data

Description of key information

ACUTE TOXICITY: VIA THE ORAL ROUTE
The key study reports an LD0 in the rat of ≥2000 mg/kg bodyweight. Both supporting studies report an LD50 in the rat of >5000 mg/kg bodyweight.
ACUTE TOXICITY: VIA INHALATION ROUTE
The 4 hour LC50 in the rat was >5.53 mg/L.
ACUTE TOXICITY: VIA DERMAL ROUTE
This study was waived on the basis that the oral and inhalation routes of administration were considered more relevant when considering the physical nature and use pattern of the substance.

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:
7 January 1987 - 22 January 1987
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
see below
Principles of method if other than guideline:
The acclimation period prior to the administration of the test material is not specified; the protocol states that it was a minimum of 48 hours. This deviation was not considered to have had an impact on the outcome of the study.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Ico rats: OFA.SD (IOPS Caw)
- Age at study initiation: 5 - 7 weeks
- Weight at study initiation (prior to fasting): 182 - 193 g (males); 170 - 185 g (females)
- Fasting period before study: yes (15 - 20 hours)
- Housing: 5 animals in stainless steel mesh cages (343 x 250 x 140 mm)
- Food consumption: ad libitum with the exception of the pre-dose fast and 3 - 4 hours following the administration of the test material.
- Water consumption: ad libitum
- Acclimation period: minimum of 48 hours

ENVIRONMENTAL CONDITIONS:
- Temperature: 22 ± 3 °C
- Humidity: 30 - 70 %
- Photoperiod: 12 hours light-dark cycle was maintained (photoperiod 0730 to 1930).

IN-LIFE DATES: From: 7 January 1987 To: 22 January 1987
Route of administration:
oral: gavage
Vehicle:
other: 10 % aqueous dispersion of gum arabic
Details on oral exposure:
VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

Preparation of the suspension: the powdered test material was put in a 20 % (w/v) suspension made immediately prior to administration in a 10 % (w/v) aqueous dispersion of gum arabic.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals per sex per dose.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
> behaviour and mortality: 15 minutes after administration of the test material, then after 1, 2 and 4 hours. Daily observations were then made for the 14 day duration of the observation period.
> weighing: one day before treatment, and on days 1, 8 and 15 after treatment
- Necropsy of survivors performed: yes
Sex:
male/female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed in rats dosed with 2000 mg/kg bw.
Clinical signs:
No clinical signs were observed.
Body weight:
No abnormal effects on bodyweight were observed.
Gross pathology:
No gross abnormalities were observed at necropsy.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD0 (males and females) was ≥ 2000 mg/kg bw and therefore the test material is not classified for acute oral toxicity based on the results of this study in accordance with EU criteria.
Executive summary:

An acute oral toxicity limit test was conducted to assess the test material in accordance with the standardised guideline OECD 401.

Groups of fasted, 5 - 7 week old Sprague-Dawley rats (5 per sex) were given a single oral dose of the test material administered as a 20 % suspension (w/v) in an aqueous dispersion of 10 % (w/v) of gum arabic at a dose of 2000 mg/kg bw and observed for 14 days.

No mortality and no clinical signs were observed during the study. No gross abnormalities were observed at necropsy.

The oral LD0 (males and females) was ≥ 2000 mg/kg bw and therefore the test material is not classified for acute oral toxicity based on the results of this study in accordance with EU criteria.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
Two supporting studies are available. The first was conducted in accordance with the standardised FHSA guideline 16 CFR 1500.3. It was well reported and adhered to sound scientific principles. It was awarded a reliability score of 1 in accordance with the criteria of Klimisch (1997).
The second supporting study is handbook data and as such has been awarded a reliability score of 2 in accordance with the criteria of Klimisch (1997).

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:
14 September 2012 - 2 October 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: RccHan:WIST
- Age at study initiation: approximately 8 - 12 weeks
- Weight at study initiation: 200 - 350 g
- Fasting period before study: no
- Housing: The animals were housed in groups of up to three by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes and provided with environmental enrichment items: wooden chew blocks and cardboard “fun tunnels”.
- Diet (e.g. ad libitum): With the exception of the exposure period, free access to food was allowed.
- Water (e.g. ad libitum): ad libitum with the exception of the exposure period.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): at least 15 per hour
- Photoperiod (hrs dark / hrs light): the lighting was controlled to give twelve hours continuous light and twelve hours darkness.

IN-LIFE DATES: From: 14 September 2012 To: 2 October 2012
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
clean air
Details on inhalation exposure:
ATMOSPHERE GENERATION
A dust atmosphere was produced from the test material using an SAG 410 Solid Aerosol Generator located adjacent to the exposure chamber. The SAG 410 was connected to a metered compressed air supply.
A particle separator was introduced before the aerosol entered the exposure chamber in order to remove large particles and thereby increase the inhalable portion of the generated aerosol.
Compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters before it was introduced to the SAG 410.
The cylindrical exposure chamber had a volume of approximately 30 litres (dimensions: 28 cm diameter x 50 cm high). The concentration within the chamber was controlled by adjusting the test material feed rate from the SAG 410. The extract from the exposure chamber passed through a ‘scrubber’ trap and was connected with a high efficiency filter to a metered exhaust system. The chamber was maintained under negative pressure.

Prior to the start of the study, test material atmospheres were generated within the exposure chamber. During this characterisation period test material input rates and the generation system were varied in order to achieve the required atmospheric conditions.

EXPOSURE PROCEDURE
Prior to the day of exposure each rat was acclimatised (for approximately 2 hours) to a tapered polycarbonate restraining tube. During the day of exposure, each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber ‘O’ ring.
A target concentration of 5.0 mg/L was used for the exposure. The mean achieved concentration was 111 % of target.

EXPOSURE CHAMBER TEMPERATURE AND RELATIVE HUMIDITY
The temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter located in a vacant port in the animals’ breathing zone of the chamber and recorded every thirty minutes.

EXPOSURE CHAMBER OXYGEN CONCENTRATION
Oxygen levels within the exposure chamber were measured by an electronic oxygen analyser located in a port in the animals' breathing zone. The test atmosphere was generated to contain at least 19 % oxygen.
The MMAD was 2.04 µm, resulting in an inhalable fraction (% <4 µm) of 82.3. The geometric standard deviation was 2.08.

EXPOSURE CHAMBER ATMOSPHERE CONCENTRATION
The actual chamber concentration was measured at regular intervals during the exposure period. The gravimetric method used glass fibre filters placed in a filter holder. The holder was temporarily sealed in a vacant port in the exposure chamber in the animals’ breathing zone and a suitable, known volume of exposure chamber air was drawn through the filter using a vacuum pump.
Each filter was weighed before and after sampling in order to calculate the weight of collected test material. The difference in the two weights, divided by the volume of atmosphere sampled, gave the actual chamber concentration.
The nominal chamber concentration was calculated by dividing the mass of test material used by the total volume of air passed through the chamber.
The nominal concentration is 257 % of the actual mean achieved atmosphere concentration and shows that keeping the aerosol airborne was relatively simplistic.

PARTICLE SIZE DISTRIBUTION
The particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period using a Marple Personal Cascade Impactor. This device consisted of six impactor stages (8.6, 5.5, 3.8, 1.7, 0.86 and 0.41 µm cut points) with stainless steel collection substrates and a back up glass fibre filter, housed in an aluminium sampler. The sampler was temporarily sealed in a sampling port in the animals’ breathing zone and a suitable, known volume of exposure chamber air was drawn through it using a vacuum pump.
The collection substrates and backup filter were weighed before and after sampling and the weight of the test material, collected at each stage, calculated by the difference.
The mean amount for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than 8.6, 5.5, 3.8, 1.7, 0.86 and 0.41 µm was calculated.
The resulting values were converted to probits and plotted against Log10 cut-point size. From this plot, the Mass Median Aerodynamic Diameter (MMAD) was determined (as the 50 % point) and the geometric standard deviation was calculated. In addition the proportion (%) of aerosol less than 4 µm (considered to be the inhalable fraction) was determined.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
The test atmosphere was sampled seventeen times during the exposure period and the actual concentration of the test material calculated.
Duration of exposure:
4 h
Concentrations:
5.53 mg/L
No. of animals per sex per dose:
3 animals per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: all animals were observed for clinical signs at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, one hour after termination of exposure and subsequently once daily for up to 14 days. Any evidence of overt toxicity was recorded at each observation.
- Frequency of observations and weighing: Individual bodyweights were recorded on arrival, prior to treatment on the day of exposure and on Days 1, 3, 7 and 14.
- Necropsy of survivors performed: yes. At the end of the 14 day observation period the animals were killed by intravenous overdose of sodium pentobarbitone. All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded. The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.53 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
There was no mortality during the study.
Clinical signs:
In addition to the observations considered to be due to the restraint procedure (such as hunched posture, pilo-erection, wet fur), increased respiratory rate was noted in all animals during exposure, on removal from the chamber and one hour post-exposure.
One day after exposure, all animals exhibited increased respiratory rate and hunched posture. In addition, two male animals also exhibited pilo-erection. Animals recovered to appear normal from Days 6 to 9 post-exposure.
Body weight:
One male and one female animal exhibited a slight bodyweight loss or showed no bodyweight gain on the first day post-exposure.
All male animals exhibited reasonable bodyweight gains during the remainder of the observation period.
In contrast, all female animals exhibited bodyweight losses or showed no bodyweight gain from Days 1 to 3; no bodyweight gain was noted in one female from Days 3 to 7.
Bodyweight gains were noted in all females during the final week of observation.
Gross pathology:
No macroscopic abnormalities were detected at necropsy.

Table 1: Exposure Chamber Atmosphere Concentrations

Duration of Exposure (minutes)

Net Weight of Sample (mg)

Volume of Air Sampled (L)

Chamber Flow Rate (L/min)

Atmosphere Concentration (mg/L)

5

10.09

2

60

5.05

15

9.49

2

60

4.75

30

10.04

2

60

5.02

45

10.23

2

60

5.12

60

12.33

2

60

6.17

75

10.52

2

60

5.26

90

11.95

2

60

5.98

105

11.32

2

60

5.66

120

9.78

2

60

4.89

135

12.74

2

60

6.37

150

11.50

2

60

5.75

165

11.11

2

60

5.56

180

10.97

2

60

5.49

198

11.13

2

60

5.57

210

11.95

2

60

5.98

225

11.42

2

60

5.71

237

11.28

2

60

5.64

Mean achieved atmosphere concentration: 5.53 mg/L (standard deviation 0.46)

 

Nominal Concentration

-Test material used: 216 g

-Air flow: 60 L/min

-Total generation time: 254 minutes*

-Nominal concentration: 14.2 mg/L

 

*Test atmospheres were generated for a total of 14 minutes prior to animal insertion to ensure that the test material concentration was being achieved.

 

Table 2: Particle Size Distribution - Cascade Impactor Data

Impactor Stage Number

Cut Point (µm)

Amount Collected (mg) Per Sample Number

Mean Amount Collected (mg)

1

2

3

3

8.6

0.08

0.06

0.05

0.06

4

5.5

0.10

0.35

0.08

0.18

5

3.8

1.03

1.40

0.97

1.13

6

1.7

1.47

2.00

1.21

1.56

7

0.86

0.64

0.53

0.61

0.59

8

0.41

0.44

0.20

0.28

0.31

Back-up filter

<0.41

0.28

0.10

0.09

0.16

Total mean amount of test material collected: 3.99 mg

 

Table 3: Particle Size Distribution - Calculation

Cut Point (µm)

Log10 Cut Point

Mean Cumulative Amount Less Than Cut Point

(mg)

(%)

Probit

8.6

0.935

3.93

98.5

7.17

5.5

0.740

3.75

94.0

6.55

3.8

0.580

2.62

65.7

5.40

1.7

0.230

1.06

26.6

4.37

0.86

-0.066

0.47

11.8

3.81

0.41

-0.387

0.16

4.01

3.25

MMAD: 2.04 µm

Geometric standard deviation: 2.08

Predicted amount <4 µm: 82.3 %

 

Table 4: Individual Bodyweights

Animal Number and Sex

Bodyweight (g) on Day:

Increment (g) During Days:

-6

0

1

3

7

14

-6 to 0

0 to 1

1 to 3

3 to 7

7 to 14

1 Male

237

269

270

276

295

321

32

1

6

19

26

2 Male

226

266

267

277

299

337

40

1

10

22

38

3 Male

220

258

256

264

280

306

38

-2

8

16

26

4 Female

215

243

245

238

243

254

28

2

-7

5

11

5 Female

196

215

215

210

219

231

19

0

-5

9

12

6 Female

191

206

208

208

208

210

15

2

0

0

2

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The 4 hour LC50 was determined to be >5.53 mg/L, therefore the test material requires no classification under the conditions of this study in accordance with EU criteria.
Executive summary:

An acute inhalation study was conducted to assess the toxicity potential of the test material in accordance with the standardised guideline OECD 436.

Male and female RccHan:WIST strain rats (3 per sex) were exposed (nose only) for 4 hours to a dust atmosphere containing the test material at a mean concentration of 5.53 mg/L, with a MMAD of 2.04 µm.

Following exposure, the animals were observed for 14 days for signs of mortality and toxicity. At the end of the observation period, all animals were subjected to necropsy.

No deaths occurred throughout the study and the 4 hour LC50 was therefore determined to be >5.53 mg/L. The test material requires no classification under the conditions of this study in accordance with EU criteria.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The key study was conducted in line with GLP and in accordance with the standardised guideline OECD 436. It was well reported and adhered to sound scientific principles. It was awarded a reliability score of 1 in accordance with the criteria of Klimisch (1997).

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Acute toxicity: via the oral route

The key study is an acute oral toxicity limit test conducted in accordance with the standardised guideline OECD 401.

Groups of fasted, 5 - 7 week old Sprague-Dawley rats (5 per sex) were given a single oral dose of the test material administered as a 20 % suspension (w/v) in an aqueous dispersion of 10 % (w/v) of gum arabic at a dose of 2000 mg/kg bw and observed for 14 days.

No mortality and no clinical signs were observed during the study. No gross abnormalities were observed at necropsy.

The oral LD0 (males and females) was ≥ 2000 mg/kg bw and therefore the test material is not classified for acute oral toxicity based on the results of this study in accordance with EU criteria.

 

The first supporting study is an acute oral toxicity limit test conducted in accordance with FHSA guideline 16 CFR 1500.3.

Groups of fasted Sprague-Dawley rats (5 per sex) were given a single oral dose of the test material in distilled water as a 50 % w/w solution at a dose of 5000 mg/kg bw and observed for 14 days.

The oral LD50 (males and females) was > 5000 mg/kg bw and therefore the material is not classified for acute oral toxicity according to EU criteria on the basis of this study.

 

The second piece of supporting information is handbook data. This quotes the oral LD50 in the rat as > 5000 mg/kg bw and therefore the test material is not classified for acute oral toxicity based on this result according to EU criteria.

 

Acute toxicity: via the inhalation route

The key acute inhalation study was conducted in accordance with the standardised guideline OECD 436.

Male and female RccHan:WIST strain rats (3 per sex) were exposed (nose only) for 4 hours to a dust atmosphere containing the test material at a mean concentration of 5.53 mg/L, with a MMAD of 2.04 µm.

Following exposure, the animals were observed for 14 days for signs of mortality and toxicity. At the end of the observation period, all animals were subjected to necropsy.

No deaths occurred throughout the study and the 4 hour LC50 was therefore determined to be > 5.53 mg/L. The test material requires no classification under the conditions of this study in accordance with EU criteria.


Justification for selection of acute toxicity – oral endpoint
The key study was conducted in line with GLP and in accordance with the standardised guideline OECD 401. It was well reported and adhered to sound scientific principles. It was awarded a reliability score of 1 in accordance with the criteria of Klimisch (1997).

Justification for selection of acute toxicity – inhalation endpoint
Only one study available.

Justification for selection of acute toxicity – dermal endpoint
In accordance with the column 2 adaptation of REACH Annex VIII, the acute dermal toxicity study (required in section 8.5.3) is not considered scientifically justified as the oral and inhalation exposure routes are the most appropriate to assess the acute toxicity hazard presented by the substance based on its physico-chemical characteristics and use pattern. Furthermore, dermal exposure of the substance is considered unlikely.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the test material does not require classification for acute oral or acute inhalation toxicity.