Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 - 24 November 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): 4,4'-(9H-fluoren-9-ylidene)bis(2-chloroaniline)
- Substance type: White powder
- Physical state: powder
- Purity: 97.3%
- Lot/batch No.: DALA101444 (8601054-003; Jet-Milled Material)
- Expiration date of the lot/batch: 18 April 2016
- Storage condition of test material: At room temperature in the dark
- Purity/composition correction factor required: No
- Test substance handling: No specific handling conditions required
- Stability at higher temperatures: Yes, maximum temperature: 300 °C

Test animals

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: Young adult animals (approximately 10 weeks old) were selected.
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Housing: Group housing of three animals per sex per cage in labelled Makrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance.
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance.
- Acclimatization period: at least 5 days before start of treatment under laboratory conditions.
- Health inspection: A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health.

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES: 10 - 24 November 2014

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
The design of the exposure chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of three animal sections with eight animal ports each. Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained at a slight negative pressure.
Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
4 h
Concentrations:
3 mg/L
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Test substance preparation: The test substance was used as delivered by the sponsor.

The test substance was administered as an aerosol by inhalation for 4 hours to two groups of three male and three female Wistar rats each. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15 and at death. Macroscopic examination was performed on the day of death or after terminal sacrifice (day 15).

The actual time weighted mean concentration was 3 ± 0.1 mg/L. The nominal concentration (amount of test substance used divided by the volume of pressurized air used) was 44 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 8%.

The concentration measurements (n=15) equally distributed over time showed that it was difficult to maintain a stable level at this technically maximum attainable concentration. The generation was interrupted in order to remove test substance deposits from the system at four occasions with a total interruption time of approximately 30 minutes. The generation time was elongated (total generation time 253 minutes) to compensate for these interruptions, resulting in an actual exposure time of at least 227 minutes. Since the interruptions and variations in concentration were taken into account by calculation a time weighted mean concentration, it was considered that the results of this study are representative for a single exposure of 3 mg/L for 4 hours. This was the technically maximum attainable concentration.
Statistics:
No statistical analysis was performed.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 3 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortality occurred.
Clinical signs:
other: Rapid breathing was seen for the animals during exposure. No clinical signs were noted following the exposure.
Body weight:
Body weight loss was seen for all females during the first week post-exposure (up to Day 8). Two females regained weight during the second week resulting in an overall body weight gain during the complete observation period, while one female remained below the initial body weight at the start of the study. Overall body weight gain in males was within the range expected for rats of this strain and age used in this type of study.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
The LC50, 4h value of 4,4’-(9H-fluoren-9-ylidene)bis(2-chloroaniline) in Wistar rats value was established to exceed the maximum obtainable concentration of 3 mg/L.
Based on these results 4,4’-(9H-fluoren-9-ylidene)bis(2-chloroaniline) does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).