Reaction products of 12-hydroxyoctadecanoic acid with ethane-1,2-diamine and hexane-1,6-diamine and 1,3-phenylenedimethanamine

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2013-09-24 to 2013-12-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of 12-hydroxyoctadecanoic acid with ethane-1,2-diamine and hexane-1,6-diamine and 1,3-phenylenedimethanamine

EC Number:

700-825-2

Molecular formula:

Not applicable (UVCB substance).

IUPAC Name:

Reaction products of 12-hydroxyoctadecanoic acid with ethane-1,2-diamine and hexane-1,6-diamine and 1,3-phenylenedimethanamine

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan (UK) Ltd.
- Age at study initiation: 10 weeks
- Weight at study initiation: 247 to 277 g (males) and 176 to 200 g (females)
- Fasting period before study:no
- Housing: The animals were housed three of one sex per cage
- Diet: Ad libitum, standard rodent diet (Rat and Mouse n°1 Maintenance Diet). This diet contained no added antibiotic or other chemotherapeutic or prophylactic agent.
- Water: Ad libitum, potable water taken from the public supply.
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19 to 23°C
- Humidity: 40 to 70%
- Air changes (per hr):Each animal room was supplied with filtered fresh air, which was passed to atmosphere and not re-circulated.
- Photoperiod (hrs dark / hrs light): 12 h continuous light and 12 h continuous dark/24 h.

Administration / exposure

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: flow through nose only chamber. This system was an aluminium alloy construction comprising a base unit, a single animal exposure section with 20 ports and a top section incorporating a central aerosol inlet.
- Method of holding animals in test chamber: Rats were held in polycarbonate tubes with their snouts protruding from the end of the tubes into the exposure chamber.
- Source and rate of air: From in-house compressed air system(breathing quality), Generator flow: 19 L/minute
- System of generating particulates/aerosols:Rotative Brush Generator mechanism designed to produce and maintain test atmospheres
containing dust by suspending material brushed from the surface of a lightly compressed powder in a stream of dry air. The concentration of dust in the air was altered by selecting different size pistons and by changing the delivery rate. A regulated flow of compressed air conducted the aerosol to the inhalation chamber.
- Temperature, humidity, pressure in air chamber:
Chamber air temperature was measured using an electronic thermometer probe placed in the breathing zone of the animals via an unused exposure port. The mean chamber temperature value was 23.4°C.
- Humidity: Chamber relative humidity was measured using an electronic hygrometer probe inserted into the breathing zone of the animals via an unused exposure port. The mean chamber relative humidity was 38.8%
Pressure in air chamber was not reported.
- Air flow rate: Airflow were 19 L/minute.
- Method of particle size determination: Particle size analysis of generated atmospheres was performed using a 8-stage cascade impactor (Marple 298). A measured volume of air was drawn at a rate of 2 litres/minute from an unused exposure port on the exposure chamber through the cascade impactor and measured using a wet-type gas meter in line with a pump. Three samples were collected during the exposure, however the first
sample was not calculated as the total capture was below the pre-determined minimum capture due to generation problems around the time of the sample. The collection substrates and the backup filter were weighed before and after sampling and the weight of test item, collected at each stage, was calculated by this difference. The total amount collected 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 < 0.52, 0.93, 1.55, 3.5, 6.0 , 9.8, 14.8 and 21.3 µm (aerodynamic diameter) was calculated. From this data, the Mass Median Aerodynamic Diameter (MMAD), and Geometric Standard Deviation (GSD) were calculated. The mean MMAD was 2.5 µm and the geometric standard deviation (GSD) was 2.37.
- Treatment of exhaust air: Extract airflow was drawn by in-house vacuum system at a flow rate of 20 L/minute. the airflow was filtered locally.

TEST ATMOSPHERE
- Brief description of analytical method used: A measured volume of air was drawn at a rate of 2 litres/minute from an unused exposure
port on the exposure chamber through a glass microfibre filter, mounted in an open face filter holder and measured using a wet-type gas meter in line with a pump. Thirteen samples were collected during the exposure. The filters were weighed before and after sampling.. The difference in the pre- and post-sampling weights, divided by the volume of atmosphere sampled, was equal to the actual achieved test atmosphere concentration. The mean achieved test atmosphere concentration was 5.14 +/- 1.66 mg/L

- Samples taken from breathing zone: yes, samples were collected from a vacant animal exposure port.

VEHICLE (if applicable)
- none

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Target concentration: 5.0 mg/L
Mean achieved atmosphere concentration: 5.14 mg/L ; Standard deviation: 1.66

No. of animals per sex per dose:

3/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d

OBSERVATIONS:
- Morbidity/Mortality: Animals were checked hourly during exposure, immediately following exposure and then at 1 h and 2 hrs post-exposure. During the 14-day observation period, the animals were observed twice daily (early and late in the working day) for morbidity and/or mortality.

- Clinical Signs: All animals were observed for clinical signs at hourly intervals during exposure, as soon as practically possible following removal from restraint at the end of exposure, 1 h and 2-hrs after exposure and subsequently twice daily for 14 d.

- Bodyweight: Individual bodyweights were recorded prior to treatment, on the day of exposure (Day 1) prior to dosing and on Days 2, 4, 8 and 15.

- Necropsy: At the end of the 14-d observation period, the animals were euthanised by exsanguination under anaesthesia (injection of pentobarbital solution) and gross macroscopic examination was performed. All animals were subject to a gross necropsy which consisted of opening the cranial, thoracic and abdominal cavities.any macroscopic abnormalities in appareance of the organs were recorded.

Statistics:

None

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.14 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no unscheduled deaths.

Clinical signs:

other: Immediately after the exposure 2/3 females were noted to have a partially closed right eye, this remained evident in 1 female at 1 hour after the exposure. Furthermore, at 1 hour after exposure 1 female was noted to be underactive with irregular breathing

Body weight:

Bodyweight losses were evident in all males and females on the day following the exposure. This loss of bodyweight is not unusual following exposures of this duration as food and water were unavailable during the exposure period, and is therefore considered not to be test substance related.
Recovery from the bodyweight loss was observed at the next weighing occasion, Day 4, after which body weight gains continued for the remainder of the study.

Gross pathology:

The macroscopic examination performed after a single administration followed by a 14 day observation period revealed enlargement of the tracheobronchial lymph nodes in all animals.

Any other information on results incl. tables

Table 7.2.2/1: Body weights - individual and group mean values (g)

Group	Male					Female
Animal number	A1	A2	A3	Mean	SD	A4	A5	A6	Mean	SD
. Day -7	246	227	241	238	9.8	166	176	184	175	9.0
. Day 1	277	247	269	264	15.5	176	190	200	189	12.1
. Day 2	269	237	248	251	16.3	172	182	189	181	8.5
. Day 4	275	249	262	262	13.0	177	188	201	189	12.0
. Day 8	290	260	280	277	15.3	183	198	212	198	14.5
. Day 15	304	278	297	293	13.5	192	210	218	207	13.3

SD : Standard Deviation

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The LC50 of the test substance was in excess of 5.14 mg/L for male and female rats.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute inhalation toxicity according to the OECD 436 guideline and in compliance with Good Laboratory Practice.

A group of three male and three female Wistar rats was exposed, nose-only, to an atmosphere of the test item using a flow-through exposure system. The animals were exposed for four hours to a target concentration of 5 mg/L followed by a fourteen day observation period. Each animal was observed for mortality and clinical signs at hourly intervals during exposure, then one hour and two hours after exposure and at least twice a day during the 14 -day observation period. Bodyweights were recorded before treatment then on the day of exposure (day 1) and on days 2, 4,8 and 15. All surviving animals were necropsied at the end of the observation period.

The mean achieved atmosphere concentration was 5.14 mg/L and the mean mass median aerodynamic diameter was 2.5 µm. No deaths occurred during the study. Immediately after the exposure two out of three females were noted to have a partially closed right eye, this was still present in one female at one hour after the exposure. Furthermore, at one hour after exposure one female was noted to be underactive with irregular breathing, the irregular breathing persisted and remained evident at two hours after the exposure. From two hours after exposure and for the remainder of the observation period all males were considered to be clinically normal; females were considered normal from Day 2.

Bodyweight losses were evident in all males and females on the day following the exposure. Recovery from the bodyweight loss was observed at the next weighing occasion, Day 4, after which body weight gains continued for the remainder of the study.

Enlargement of the tracheobronchial lymph nodes was noted in all animals.

The 4 -hour inhalation LC50 was found to be greater than 5.14 mg/L (ie 5140 mg/m3).