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Administrative data

Description of key information

No toxicity was observed at the limit dose  of 2000 mg/kg bw upon acute oral and dermal toxicity (BSL 2009a, NOTOX 2009a). Studies were performed according to OECD guidelines 423 and 402 and following GLP.  Acute inhalation of dust caused reversible clinical symptoms but no mortality (BASF 2014, OECD 403, GLP).

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:
January - February 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline and GLP compliant study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: WISTAR RjHan : WI rats (Full-Barrier)
Sex:
female
Details on test animals and environmental conditions:
Source: Janvier, F-53940 Le Genest-Saint-Isle
Sex: female, non-pregnant, nulliparous
Number of animals: 3 per step
Age at the beginning of the study: 8 - 12 weeks old
Body weight at the beginning of the study: Animals no 1 – 3, step 1: 213 – 220 g; animals no 4 – 6, step 2: 209 – 238 g

Housing and Feeding Conditions:
- Semi-barrier in an air conditioned room
- Temperature: 22 ± 3 °C
- Relative humidity: 55 ± 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH value of approx. 2.8
(drinking water, municipal residue control, microbiologically controlled
at frequent intervals)
- The animals were kept in groups in IVC cages, type III H, polysulphone
cages on Altromin saw fiber bedding
- Certificates of food, water and bedding are filed at BSL Bioservice
- Adequate acclimatisation period of at least five days
The animals were derived from a controlled full barrier maintained breeding system (SPF). According to Art. 9.2, No.7 of the German Act on Animal Welfare the animals were bred for experimental purposes.
Route of administration:
oral: gavage
Vehicle:
other: Carboxymethylcellulose 1% in water
Details on oral exposure:
The animals were marked for individual identification by tail painting. Prior to the administration a detailed clinical observation was made of all
animals. Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing. The test item was administered at a single dose by gavage using an intubation cannula.
The test item was administered at a dosing volume of 10 mL/kg body weight.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Six
Control animals:
no
Details on study design:
All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality. The animals were weighed on day 0 (prior to the administration) and on days 7 and 14. A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as the symptoms noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded. Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, letharg , sleep and coma. At the end of the observation period the animals were sacrificed by an overdosage of pentobarbital injected intraperitoneally (Narcoren®, manufacturer: Merial; batch no.: 185118; expiry date: November 2011) at the dosage of approx. 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded. All of the animals were subjected to gross necropsy. All gross pathological changes were recorded.
Statistics:
not applicable
Sex:
female
Dose descriptor:
LD0
Effect level:
> 2 000 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No mortality occured.
Clinical signs:
No clinical signs were observed.
Body weight:
No changes in body weight occured.
Gross pathology:
No gross pathology findings were observed.
Other findings:
No other treatment-related findings were observed.
Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Migrated information
Conclusions:
No indication of toxicity was observed after application of a single oral dose of 2000 mg/kg bw. The test item is therefore considered to be of low acute oral toxicity.
Executive summary:

All animals survived a single bolus dose of 2000 mg/kg bw. No adverse signs were noted for clinical observations, body weight and gross pathology.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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:
2013-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and OECD guideline compliant study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
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
- Source: Harlan Laboratories B.V., 5961 NM Horst, The Netherlands
- Age at study initiation: male animals approx. 9 weeks, female animals approx. 11 weeks
- Weight at study initiation: Animals of comparable weight (± 20% of the mean weight)
- Fasting period before study:
- Housing: Single housing
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 – 24°C
- Humidity (%): 30 – 70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2013-12-02 To: 2013-12-18
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Head nose inhalation system INA 10 (glass steel construction)
- Exposure chamber volume: 34L
- Method of holding animals in test chamber: restraining tubes
- Source and rate of air: Air passed through activated charcoal and a HEPA filter. Flow rates: 0.8 m³/h (during the first 2 hours), 1.0 m³/h (during hour 3), 1.3 m³/h (during hour 4)
The flow was adjusted and continuously measured with a flowmeter (Yokogawa).
During technical trial, an atmospheric concentration of slightly higher than 5 mg/L had been achieved using a supply air flow of 0.8 m³/h and an exhaust air flow of 0.6 m³/h. During the study however, the measured concentrations were below 5 mg/L, probably due to the presence of animals. In order to achieve a higher atmospheric concentration, the supply air flow was increased to 1.0 mg/m³ in hour 3, and subsequently increased to 1.3 m³/h.

- Method of conditioning air: Dosing-wheel dust generator
- System of generating particulates/aerosols: The test substance was des-agglomerated in a mixer (mixing for 10 seconds) under addition 1% AEROSIL® 200 and 1% AEROSIL® R 972 before introduction into the dust generator.) The dust was produced inside the dust pre-chamber using a
dosing wheel dust generator and compressed air. The dust was passed via the cyclonic separator into the inhalation system.

- Method of particle size determination: Stack Sampler Mark III (Andersen).
Before sampling, to reduce rebounding effect, the impactor stages were coated with parafine oil. Thereafter, the impactor were assembled with preweighed glass fiber collecting discs, and equipped with a backup particle filter. The impactor was connected to the vacuum pump and two samples were taken from the breathing zone of the animals sampling occurred 30 minutes (or later) after the beginning of the exposure.
The sample volume for each sample was 6 L.

- Treatment of exhaust air: The exhaust air was filtered and conducted into the exhaust air of the building.
- Temperature, humidity, pressure in air chamber: 21.4 °C, 25.2% humidity. Lower amount of exhaust air, which was adjusted by means of a separate exhaust air system, achieved a positive pressure inside the exposure system. This ensured that the mixture of test substance and air was not diluted with laboratory air in the breathing zones of the animals

TEST ATMOSPHERE
- Brief description of analytical method used: Preweighed filters were placed into the filtration equipment (MN 85/90 BF (d = 47 mm)). By means of the vacuum pump metered volumes of the dust were drawn through the filter. For each sample the dust concentration in mg/L was calculated from the difference between the preweight of the filter and the weight of the filter after sampling, with reference to the sample volume of the inhalation atmospheres. Mean and standard deviation were calculated for the concentration from the results of the individual measurements. The concentration was corrected for the amount of additive used. Samples were taken every hour from a total sample volume of 6L air.

- Samples taken from breathing zone: yes

VEHICLE
- Composition of vehicle (if applicable): To improve the flowability, 1 % AEROSIL® 200 was added to the test substance. In addition 1% AEROSIL® R 972 was added as an antihydroscopic agent.


TEST ATMOSPHERE
- Particle size distribution: See figure

Sample MMAD (µm) Geometrical standard deviation
1 4.4 3.3
2 2.8 2.3


CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: The limit dose was chosen because the substance was found to be non-toxic in all available toxicity studies.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Gravimetric measurement
Duration of exposure:
4 h
Concentrations:
4.651 mg/L +/- 0.581 (measured; >5 mg/L nominal)
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: Individual body weights once during the acclimatization period, shortly before exposure (day 0) and at least on days 1, 3 and 7, and before the sacrifice of the animals at the end of the observation period. 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.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Statistics:
not required
Sex:
male/female
Dose descriptor:
LC0
Effect level:
>= 4.651 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4.651 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
none
Clinical signs:
other: See table 1
Body weight:
The mean body weights of the animals decreased during the first post exposure observation day and increased from study day 3 onward.
Gross pathology:
No adverse findings were observed.

Table 1: Clinical signs

Test group (4.651 mg/L)

Male animals

Female animals

Fur, yellow discolored

d6 - d14

d5 – d14

Fur, substance contaminated

d0 – d11

d0 – d4

Respiration, intermittent

d0 – d1

d0 – d4

Respiration, labored

h2 – h4

h2 – h4

Piloerection

d0 – d7

d0 – d5

Table 2: Concentration in the test chamber

time point of sampling measured concentration
1h 4.06
2h 4.246
3h 5.101
4h 5.196
Mean 4.6508
Mean (rounded) 4.651
Standard deviation of the mean 0.581
Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
No mortality was observed in rats exposed to a dust at a measured concentration of 4.651 mg/L for 4hours.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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:
2009-03-11, 209-03-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Performed according to OECD guideline and under GLP
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF Guidelines (2000), including the most recent revisions
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approx. 8 weeks
- Weight at study initiation: did not exceed +/- 20% of the sex mean
- Housing: Individually in labeled Macrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet: ad libitum, pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: ad libitum, tap water
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5 – 20.7
- Humidity (%): 41 - 65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12
Type of coverage:
occlusive
Vehicle:
CMC (carboxymethyl cellulose)
Details on dermal exposure:
TEST SITE
- Area of exposure: back
- % coverage: approx. 10 %
- Type of wrap if used: the test substance formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with tap water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg (10 mL/kg) body weight
- Constant volume or concentration used: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 1 % aqueous carboxymethyl cellulose
Duration of exposure:
24 hours
Doses:
1
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: twice daily (mortality/viability); days 1 (pre-administration), 8 and 15 (body weights)
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs
Statistics:
Observations/measurements in the study were recorded electronically using the following programme(s):
REES Centron Environmental Monitoring system version SQL 2.0 (REES scientific, Trenton, NJ, USA);
TOXDATA version 8.0 (NOTOX B.V., ‘s-Hertogenbosch, The Netherlands): Clinical signs, Body weights
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No mortality occurred
Clinical signs:
Hunched posture, piloerection, chromodacryorrhoea and/or hypothermia were noted among animals on Days 1 and/or 2.
Scales were observed in the treated skin-area of one animal during the observation period.
Yellow staining of the treated skin-area was noted for all animals during the observation period. This was considered related to the staining properties of the test substance and therefore of no toxicological relevance.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Two groups, each of three female WISTAR RjHan: WI (SPF) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight (BSL 2009a). The study followed OECD testing guideline 423 and the principles of GLP. The test item was suspended in a vehicle (CMC 1%) at a concentration of 0.2 g/mL and administered at a dosing volume of 10 mL/kg. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 0 (prior to the administration) and on days 7 and 14. All animals were necropsied and examined macroscopically. No signs of toxicity were recorded in any of the animals. Throughout the 14-day observation period, the weight gain of the animals was within the expected range. No test item related findings were recorded at necropsy.

The test substance was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours (NOTOX 2009a). The study followed OECD testing guideline 402 and the principles of GLP. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). No mortality occurred. Hunched posture, piloerection, chromodacryorrhoea and/or hypothermia were noted among animals on Days 1 and/or 2. Scales were observed in the treated skin-area of one animal during the observation period. Yellow staining of the treated skin-area was noted for all animals during the observation period. This was considered related to the staining properties of the test substance and therefore of no toxicological relevance. The body weight gain during the observation period was within the range expected for rats used in this type of study. No abnormalities were found at macroscopic post mortem examination of the animals.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for acute oral, inhalation or dermal toxicity under Directive 67/548/EEC, as amended for the 31st time in Directive2009/2/EG.

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for acute oral, inhalation or dermal toxicity under Regulation (EC) No. 1272/2008, as amended for the seventh time in Regulation (EC) No 2015/1221.