Registration Dossier

Diss Factsheets

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no

Test material

Constituent 1
Reference substance name:
Fatty acids, C8-18 and C18-unsatd., reaction products with diethanolamine and propylene oxide
EC Number:
600-026-8
Cas Number:
1000817-22-0
Molecular formula:
Unspecified
IUPAC Name:
Fatty acids, C8-18 and C18-unsatd., reaction products with diethanolamine and propylene oxide
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): Kerocom FM 38
- Physical state: liquid
- Analytical purity: 100% (UVCB) Information of the comparable substance No. 08/0812-2 (see analytical report 11L00413)
- Lot/batch No.: 24B187
- Expiration date of the lot/batch: Expiry date: 23 Mar 2016
- Stability under test conditions: The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
- Storage condition of test material: Room temperature; avoid temperatures <0°C and >40°C

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories B.V.
- Age at study initiation: Young adult animals (male animals approx. 8 weeks, female animals approx. 10 weeks)
- Weight at study initiation: Animals of comparable weight
- Housing: Single housing or up to 5 animals
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Acclimatization for at least 5 days before exposure.

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

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF THE INHALATION ATMOSPHERE
Test-substance preparation: The test substance was dosed unchanged.
Equipment: Two-component atomizer Mod. 970 (stainless steel, Schlick) Continuous infusion pumps PHD Ultra (Harvard Apparatus, Inc., Holliston, Massachusetts, U.S.A.)
Generation technique: Liquid aerosols were generated. For each test group the aerosols were produced by continuously pumping amounts of the test substance to a two-component atomizer. Using compressed air, the aerosol was produced with the atomizer inside the exposure system.

EXPOSURE
Nose-only inhalation system INA 20 (glass-steel construction, BASF SE, volume V 55 L):
the animals were restrained in glass tubes and their snouts projected into the inhalation system.
The homogenous distribution of test substance atmosphere/s in this inhalation system has been verified with model aerosols.
Conditioned air:
The central air conditioning system provides cold air of about 15°C. This cold air passes through an activated charcoal filter, is adjusted to room temperature of 20 to 24°C and passes through a second particle filter (H13 (HEPA) Camfil Farr, Germany). The so generated conditioned air was used to generate inhalation atmospheres.
Compressed air:
Compressed air was produced by an oil-free compressor (HT 6, Josef Mehrer GmbH & Co KG, Germany). For this purpose, air is filtered by an inlet air strainer and introduced into the compressor. After passing through a second ultra-filter (SMF 5/3, 108 mm, Donalson), the compressed air (15 bar) is stored in a storage of 1500 or 5000 L. The compressed air is conducted to the laboratories via pipes, where the pressure is reduced to 6 bar. In the laboratory, the compressed air can be taken as required.
Exhaust air:
The exhaust air was filtered and conducted into the exhaust air of the building. The exposure system was located inside an exhaust cabin in an air-conditioned laboratory. During each exposure, the following scheduled parameters were recorded four times at about
1-hour intervals:
Supply air flows (compressed air): 1.5 m³/h (from a central air-conditioning system)
The flows were adjusted and continuously measured with a flowmeter (Yokogawa).

Exhaust air flows: 1.35 m³/h (test group 1); 1.3 m³/h (test group 2)
The flows were adjusted and continuously measured with a flowmeter (Yokogawa).

The lower amounts of exhaust air, which were adjusted by means of a separate exhaust air system, achieved positive pressures inside the exposure systems. This ensured that the mixtures of test substance and air were not diluted with laboratory air in the breathing zones
of the animals.
Air changes of about 27 times per hour can be calculated by dividing the supply air flows by the volume of the inhalation systems.

The animals were exposed to the inhalation atmosphere for 4 hours plus equilibration time of the inhalation systems (t99 about 10 min).
No surveillance of the oxygen content in the inhalation system was performed. The air change was judged to be sufficient to prevent oxygen depletion by the breathing of the animals, and the concentration of the test substance used could not have a substantial influence on oxygen partial pressure.
Temperatures: The temperatures in the inhalation system were measured continuously with a digital thermometer.
Relative humidities: The relative humidities in the inhalation system were measured with a dielectric probe (Testo).
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
1.028 mg/L and 5.30 mg/L
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: A check for the presence of feed and drinking water was made twice a day on workdays and once daily on weekends and public holidays. 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. Additionally, body weight was measured in animals that died from study day 1 onwards. Clinical observations were recorded for each animal before exposure, separately several times during exposure (usually
hourly) and after exposure. At least once daily on the preexposure day and during the post exposure observation period. A check for any dead or moribund animal was made twice each workday and once on Saturdays, Sundays and on public holidays.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,histopathology
Statistics:
Detailed statistical analysis were conducted

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.3 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
Test group 1 (1.028 mg/L):
No lethality was observed in male and female animals during the study period of 14 days.
Test group 2 (5.30 mg/L):
No lethality was observed in male animals and one of the five females died on study day 5 during the post exposure observation period.
Clinical signs:
other: 1,028 mg/L substance contaminated fur (d0-1), colorless nasal discharge (d0-3), abdominal and labored respiration (d0-6), respiration sounds (d0-6), piloerection (d0-1) No abnormalities were detected in the male animals during the post exposure observatio
Body weight:
The mean body weights of the surviving animals in test group 1 (1.028 mg/L) and test group 2 (5.30 mg/L) decreased on the first post exposure observation day and thereafter increased.
Gross pathology:
Animals that died during the study period:
Stomach, Jejunum, Cecum: dilation with gaseous content

To further evaluate the macroscopic findings, histopathological examination of the respiration
tract (nasal cavity, larynx, pharynx, trachea, lung) from female animal No. 409 which died on
study day 5 was performed.
The larynx, pharynx, trachea and the lung showed no findings.
The nasal cavity showed a moderate purulent inflammation, characterized by a neutrophilic
granulocytic exudate in the lumen at all levels. Foreign bodies were entrapped in the exudate,
most likely being the cause of the inflammation.
The stomach, jejunum and cecum showed no correlation with the gross findings. No other
findings were noted.

Applicant's summary and conclusion

Conclusions:
Under the current study conditions, the LC50 value was > 5.30 mg/L (calculated based on analytical concentration) in male and female Wistar rats after 4 hour inhalation exposure to liquid aerosol of Kerocom FM 38.