Fatty acids, C16 and C16-18-unsatd., mixed esters with adipic acid and pentaerythritol

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Jun - 06 Jul 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18 (even numbered) and C18-unsatd., adipic acid esters with pentaerythritol

EC Number:

921-836-0

Cas Number:

1190402-12-0

Molecular formula:

not available, UVCB substance

IUPAC Name:

Fatty acids, C16-18 (even numbered) and C18-unsatd., adipic acid esters with pentaerythritol

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Physical state: yellow liquid
- Analytical purity: 100%
- Composition of test material: Fatty acids, C16-18, even numbered and C18-unsatd. esters with Propylidynetrimethanol
- Lot/batch No.: 15651
- Stability under test conditions: the test substance was expected to be stable for the duration of testing.
- Storage condition of test material: at room temperature

Test animals

Species:

rat

Strain:

other: Sprgaue-Dawley-derived, albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 252-272 g (males), 180-200 g (females)
- Housing: animals were housed individually in suspended stainless steel cages with mesh floors. Litter paper was placed beneath the cage and was changed three times per week. During exposure, animals were individually housed in polycarbonate holding tubes.
- Diet: Harlan Teklad global 16% Protein Rodent Diet® #2016, ad libitum (except during exposure)
- Water: filtered tap water, ad libitum (except during exposure)
- Acclimation period: 9 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22
- Humidity (%): 54-76
- Air changes (per hr): 13
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 22 Jun 2012 To: 06 Jul 2012

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Nose-Only Inhalation Chamber (ADG Developments LTD)
- Exposure chamber volume: approx. 6.7 L
- Method of holding animals in test chamber: animals were individually housed in polycarbonate holding tubes which seal to the chamber with an “O” ring during exposure.
- Source and rate of air: filtered air supplied by an air compressor (Powerex Model: SES05) at 30L/min (measured mean air flow: 36L/min)
- System of generating particulates/aerosols: the test atmosphere was generated using a 1/4 inch JCO atomizer (Spraying Systems Co.), FC3 fluid cap (Robert Miller Associates) and 70SS air cap (Spraying Systems Co.).
- Method of particle size determination: an eight-stage ACFM Andersen Ambient Particle Sizing Sampler was used to assess the particle size distribution of the test atmosphere. Samples were withdrawn from the breathing zone of the animals at two intervals. The filter paper collection stages were weighed before and after sampling to determine the mass collected upon the stage. The aerodynamic mass median diameter and the geometric standard deviation were determined graphically using two-cycle logarithmic probit axis.
- Temperature, humidity, pressure in air chamber: 23-25 °C, 59-62%

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric samples were withdrawn at 6 intervals form the breathing zone of the animals. Samples were collected using 37 mm glass fibre filters (GF/B Whatman) in a filter holder attached by 1/4 inch Tygon tubing to a vacuum pump (GAST Model #1531-107B-G557X). Filter papers were weighed before and after collection to determine the mass collected. This value was divided by the total volume of air sampled to determine the chamber concentration. The collections were carried out for 1 min at airflows of 4 L per min. Sample airflows were measured using a Mass Flowmeter (Aalborg Model #GFC-17).
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- Particle size distribution: the particle size distribution of the test aerosol is shown in Table 1 under “Any other information on materials and methods incl. tables”).
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): individual values: 2.41 µm / 1.97 µm and 2.11 µm / 1.84 µm; mean value MMAD: 2.3 µm

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetric

Duration of exposure:

4 h

Concentrations:

50.74 mg/L (nominal concentration)
5.06 mg/L (analytical concentration)

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: on the day of and prior to exposure, the rats were examined for health. All animals were observed for mortality during the exposure period. Furthermore, the animals were examined for signs of gross toxicity, and behavioural changes upon removal from the exposure tube and at least daily thereafter during the 14-day observation period. Individual body weights were determined prior to test substance exposure on SD 0, and again on SD 1, 3, 7 and 14 (study termination).
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, other: behaviour

Statistics:

The mean value and standard deviation of the gravimetric chamber concentration was determined.

Results and discussion

Preliminary study:

Preliminary trials were performed to establish suitable generation procedures for the test aerosol in order to achieve the desired chamber concentration (5.0 mg/L) and the desired particle distribution (MMAD between 1 and 4 µm). Procedures and aerosolisation equipment used in the main experiment were based on the results of a pre-test, in which a gravimetric concentration of 5.15 mg/L and a MMAD of 2.51 of the test aerosol were achieved.

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.06 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortalities were observed during the study.

Clinical signs:

other: Immediately following exposure, all rats exhibited irregular respiration, but recovered from this symptom by SD 3 and appeared active and healthy for the remainder of the 14-day observation period (see Table 3 under "Any other information on results incl.

Body weight:

Although 9/10 rats lost body weight or failed to gain body weight by Day 1, all animals showed a continued weight gain thereafter through Day 14 of the study period.

Gross pathology:

No gross abnormalities were noted at necropsy in any of the treated animals.

Any other information on results incl. tables

Table 2. Exposure chamber atmosphere concentrations

Sample number	Sampling time [h]	Mass collected [mg]	Airflow sampled [L per min]	Collection time [min]	Chamber concentration [mg/L)
1	0.50	20.3	4	1	5.08
2	1.00	20.3	4	1	5.08
3	2.00	20.3	4	1	5.08
4	2.50	20.1	4	1	5.03
5	3.50	20.2	4	1	5.05
6	3.75	20.1	4	1	5.03
Average chamber concentration ± standard deviation					5.06 ± 0.02

Table 3. Table for acute inhalation toxicity

Target concentration [mg/L air]	Number of animals with clinical signs*	Mortality (%)	Time of death	Duration of clinical signs	Gross pathology
Males
5.06	0/5/5	0	---	CR - Day 2	no abnormalities
Females
5.06	0/5/5	0	---	CR - Day 2	no abnormalities
LC50 > 5.06 mg/L air

* first number = number of dead animals

second number = number of animals with clinical signs

third number = number of animals used

CR = removal from exposure tube

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified
DSD: not classified