2,3-Epoxypropyl neodecanoate, oligomeric reaction products with phosphorous acid

Endpoint summary

Description of key information

Under the conditions of the study in rats according to OECD 423 the acute toxicity after oral application is greater than 2000 mg/kg bw for the test item (BSL Bioservice, 2004). Furthermore, an acute inhalation toxicity study was conducted in rats according to OECD 403 revealed LC50 of greater than 5.03 mg test item/L air/4 hours (LPT, 2016). An acute dermal toxicity study is not available for the test substance.

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:

2004-07-05 to 2004-07-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

PREPARATION of the TEST ITEM
- In the first and second step 2000 mg of the test item were diluted in Cottonseed Oil (Sigma Chemicals Co., Lot 103K0064) ad 10 mL.
- The test substance was freshly mixed prior to administration and stirred troughout dose administration to guarantee stability and homogenicity.
The vehicle was chosen due to its non-toxic characteristics.

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rats (HsdBrlHan: WIST), Sex: female
- Step 1: Body weight at the commencement of the study: 150 - 153 g
Step 2: Body weight at the commencement of the study: 140 - 165 g
- Three female animals were used for each step
- The animals were derived from a controlled full barrier maintained breeding system (SPF)
- Source: Harlan Winkelmann GmbH, D-33178 Borchen
- According to Art. 9.2, No.7 of the Greman Act on Animal Welfare the animals were bred for experimental purposes.

ANIMAL HUSBANDRY
The animals were barrier maintained (semi-barrier) in an air conditioned room
- Temperature: 22 +/- 3 °C
- Rel. humidity: 55 +/- 10%
- Artificial change: 10 x / hour
- Feeding ad libitum, Altromin 1324 maintenance diet for rats and mice, totally-pathogen-free (TPF)
- Free access to tap water (drinking water, municipal residue control, microbiol. controlled periodically)
- The animals were kept in Macrolon cages on Altromin saw fiber bedding
- Certificates of food, water and bedding are filed at BSL Bioservice
- Adequate acclimatization period

PREPARATION OF THE ANIMALS
- The animals were marked for individual identification by tail painting
- Prior to the adminstration a detailed clinical observation was made of all animals
- Prior to administration of the test item animals were fasted by withholding food overnight.
Following the period of fasting the animals were weighed and the test item was administered. Then the food was withheld for a further 3-4 hours.

Route of administration:

oral: gavage

Vehicle:

cotton seed oil

Details on oral exposure:

ADMINISTRATION:
- The test item was aministered in a single dose by gavage using an intubation cannula.
- Volume of administration: The test item was administered according to body weight at a volume of 10 mL/kg bw.

Doses:

The starting dose (step 1 and 2) was selected to be 2000 mg/kg body weight. According to the acute toxic class method regime no further testing was required since no compound-related mortality was found.

No. of animals per sex per dose:

6 (step 1 and 2)

Control animals:

no

Details on study design:

OBSERVATION:
- Animals were observed for 14 days after dosing.

WEIGHT ASSESSMENT:
- The animals were weighed prior to the administration and once a week thereafter.

CLINICAL EXAMINATION:
- A careful clinical examination was made several times on the day of dosing. Part of this were at least three observations within the first four hours post-dose.
Animals were observed once a day thereafter.
- 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, lethargy, sleep and coma.

PATHOLOGY:
- At the end of the observation period

Statistics:

EVALUATION OF RESULTS:
Individual reactions of each animal were recorded at each observation time. Toxic response data were recorded by sex and dose level. Nature, severity and duration of clinical observations were described. Body weight changes were summarized in tabular form. Necropsy findings were described.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured.

Clinical signs:

No clinical signs of toxicity were observed throughout the observation period for any animals of step 1 and 2.

Body weight:

Throughout the 14-days observation period no weight loss was recorded in any animals of step 1 and 2 (see tables "Absolute body weights in g" below).
The weight gain was within the expected range.

Gross pathology:

Beside acute injection of blood vessels in the adominal region, which is due to the euthanasia injection no special gross pathological changes were found in any animals of step 1 nad 2.

Other findings:

No other treatment related effects were observed.

Absolute Body Weights in g

Step 1 (2000 mg/kg bw)

Animal No./Sex	Day 0	Day 7	Day 14
1 / female	150	167	182
2 / female	153	179	193
3 / female	151	182	203

Step 2 (2000 mg/kg bw)

Animal No./Sex	Day 0	Day 7	Day 14
1 / female	147	164	167
2 / female	140	160	162
3 / female	165	176	188

Conclusions:

Under the conditions of this study the acute toxicity after oral application is greater than 2000 mg/kg bw.

Executive summary:

In this study the acute toxic class method according to OECD 423 was performed with the test item.

In the first step the test item was given at the limit dose of 2000 mg/kg body weight to a group of 3 female rats (HsdBrlHan:WIST) as a single administration by oral gavage at the volumne of 10 mL/kg bw. In the second step the test item was given at the same dose and volume to a further group of 3 female rats (HsdBrlHan:WIST).

The dosage of 2000 mg/kg bw caused no compound-related mortality in any animals of step 1 and 2 within 14 days post-dose.

A careful cinical examination was made several times on the day of dosing. Part of this were at least three observations within the first four hours post-dose. Animals were observed once a day thereafter.

At the end of the observation period the surviving animals were sacrified. Necropsy of all animals was carried out to record gross pathological changes.

No clinical signs of toxicity were observed throughout the observation period for any animals of step 1 and 2.

Beside acute injection of blood vessels in the abdominal region, which is due to the euthanasia injection no special gross pathological changes were found in any animals of step 1 and 2.

Throughout the 14 -days observation period no weight loss was recorded in any animals of step 1 and 2. The weight gain was within the expected range.

No other treatment related effects were observed.

Therefore, under the conditions of this study the acute toxicity after oral application is greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study is a guideline study with Klimisch score 1 (reliable without restrictions).

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:

2016-03-08 to 2016-05-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted September 7, 2009

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Acute Inhalation Toxicity, published in the Official Journal of the European Union L81, dated January 24, 2014.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Rat / CD / Crl:CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ORGANISMS: 
- Strain: Rat / CD / Crl: CD(SD)
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age: males: Approx. 8 weeks, females: approx. 10 weeks
- Weight at study initiation: males: 290 - 315 g, females: 250 - 290 g
- Number of animals: 3 males and 3 females
- Fasting period before study: 16 hours
- Housing: groups of three animals
- Diet: ad libitum, Commercial diet, ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany
- Water: ad libitum, tab water
- Acclimatisation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3 °C
- Humidity (%): 55 + / - 15 %
- Photoperiod (hrs dark / hrs light): 12 hours darkness, 12 hours artifical light

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: compressed filtered air

Mass median aerodynamic diameter (MMAD):

2.585 µm

Geometric standard deviation (GSD):

2.85

Remark on MMAD/GSD:

The Geometric Standard Deviation (GSD) of the MMAD was calculated from the quotient of the 84.1%- and the 50%-mass fractions, both obtained from the above mentioned non-linear regression analysis.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Diluent: Dimethylsulfoxyd (DMSO), The test item is an emulsion with a high viscosity. Hence, a mixture of the test item with water (1 + 1, m/m) was the highest possible concentration to obtain an appropriate aerosol.
- Type of exposure: Using a dynamic inhalation chamber (air changes/h (≥ 12 times)) with a nose-only exposure of the animals according to KIMMERLE & TEPPER .
- Method of holding animals in test chamber (volume 40 l): The apparatus consists of a cylindrical exposure chamber (volume 40 L) which holds a maximum of 20 animals in pyrex tubes at the edge of the chamber in a radial position.
- Source of air: compressed filtered air
- Method of conditioning air: A manometer and an air-flow meter were used to control the constant supply of compressed air and the exhaust, respectively. Flow rates were checked hourly and corrected if necessary.
- Oxygen content: The oxygen content in the inhalation chamber was 21% v/v. It was determined at the beginning and at the end of the exposure with a DRÄGER Oxygen-analysis test set (DRÄGER Tube Oxygen 67 28 081).
- Type or preparation of particles: The aerosol of the test item was obtained using a spray-jet. The spray-jet was fed with compressed air (5.0 bar) from a compressor and with the test item using a TSE infusion pump . At the bottom of the exposure chamber, the air was sucked off at a lower rate than created by the dust generator in order to produce a homogenous distribution and a positive pressure in the exposure chamber (inflow 900 L/h, outflow 800 L/h).
- Analysis of the aerosol concentration: The actual aerosol concentration in the inhalation chamber was measured 4 times gravimetrically with an air sample filter (Minisart SM 17598; 0.45 µm) and pump (Vacuubrand, MZ 2C ) controlled by a rotameter. Aerosol samples were taken once every hour during the exposure. For that purpose, a probe was placed close to the animals' noses in the inhalation chamber and air was sucked through the air sample filter at a constant flow of air of 5 L/min for 1 minute. The filters were weighed before and after sampling on an analytical balance (accuracy 0.1 mg).
- Method of particle size distribution: An analysis of the particle size distribution was carried out twice during the exposure period using a cascade impactor according to MAY .
The impactor is a device that classifies particles present in a sample of air or gas into known size ranges. It does this by drawing the air sample through a cascade of progressively finer nozzles. The air jets from these nozzles impact on pre-weighed plane sampling surfaces (slides).
Each stage represents an aerodynamic size range and collects finer particles than its predecessor. Each successive stage represents a special aerodynamic cut off diameter.
The dust from the exposure chamber was drawn through the cascade impactor for 1 minute at a constant flow rate of 5 L/min. The slides were removed from the impactor and weighed on an analytical balance (SARTORIUS, type 1601 004, precision 0.1 mg).
The mass median aerodynamic diameter (MMAD) was estimated by means of non-linear regression analysis. The 32 µm particle size range and the filter (particle size range < 0.5 µm) were not included in the determination of the MMAD in order not to give undue weight to these values.
The Geometric Standard Deviation (GSD) of the MMAD was calculated from the quotient of the 84.1%- and the 50%-mass fractions, both obtained from the above mentioned non-linear regression analysis.
- Temperature, humidity: The temperature (22°C ± 3°C) and humidity was checked and noted once every hour during the exposure period of the experiment.

TEST ATMOSPHERE
- Concentrations: 5.03 mg/l 
Air flow entrance (L/h): 900
Air flow exit (L/h): 800
Air change (changes per hour): 31,6

VEHICLE
compressed filtered air

TEST ATMOSPHERE
see table below

CLASS METHOD (if applicable)
Before initiating the study with the animals, a pre-test was carried out with the exposure system in order to verify that under the experimental settings chosen, the limit concentration of 5 mg/L air could be achieved by gravimetric analysis.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

The actual aerosol concentration in the inhalation chamber was measured 4 times gravimetrically with an air sample filter (Minisart SM 17598; 0.45 µm) and pump (Vacuubrand, MZ 2C ) controlled by a rotameter.

Duration of exposure:

4 h

Concentrations:

The actual aerosol concentrations of 5.03 mg /L air was measured at the animals’ nose.

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

EXAMINATIONS: 
- Post dose observation period: 14 days
- body weights: Individual body weights of animals were determined 1 day before administration (acclimatisation period), on test day 1 prior to exposure and on test days 2, 4, 8 and 15.
- mortality: once per hour during, and once after treatment on day of  exposure;  thereafter twice daily
- clinical signs: . A careful clinical examination was made at least once daily until all symptoms subsided, thereafter each working day.
Cageside observations included, but were not limited to: changes in the skin and fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, as well as somatomotor activity and behaviour pattern.
Particular attention was directed to observation of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Necropsy: all animals was carried out and all gross pathological changes were recorded. In addition, the weight of the lungs was determined.

Statistics:

not necessary

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.03 mg/L air

Based on:

act. ingr.

Exp. duration:

4 h

Mortality:

No animal died prematurely.

Clinical signs:

other: Slight ataxia and slight dyspnoea immediately until 30 minutes or 3 hours after end of exposure.

Body weight:

One of the 3 female animals appeared to be reduced in body weight gain since test day 8.

Gross pathology:

None findings

Other findings:

no other findings

The mean actual exposure concentration, MMAD and GSD of the test item were as follows:

actual concentration	nominal concentration	mass median aerodynamic diameter	respirable amount particle size ≤4 µm
[mg/L air]	[mg/L air]	[µm]	[mg/L air]	[%]
5.03	27.78	2.585	2.00	39.7

Conclusions:

Under the present test conditions, LC50-value for CD rats following inhalation of the test item for 4 hours was determined as follows (actual concentration): LC50: exceeding 5.09 mg test item/L air.

Executive summary:

The aim of the present experiment was to obtain information on the acute toxicity and respiratory irritation, following a single 4-hour inhalation exposure of rats to the test item in an acute inhalation toxicity study.

Under the present test conditions, a 4-hour exposure to test item at the concentration of 5.03 mg/L air revealed slight ataxia and slight dyspnoea on test day 1 in all 3 of 3 male and 3 of 3 female animals. No animal died prematurely. One of 3 male and 1 of 3 female animals appeared to be reduced in body weight gain. No pathological findings were noted at necropsy.

LC₅₀value for males and females combined (14 days):

           > 5.03 mg test item/L air/4 hours

The test item is a viscous liquid. A 50% (m/m) concentration of the test item in dimethylsulfoxyd (DMSO) was the most suitable concentration to obtain an appropriate aerosol.

All concentrations refer to the undiluted supplied test item.

 

Aerosol concentration and particle size distribution

The generated aerosol had mass median aerodynamic diameters (MMAD) of 2.585 µm as determined with a cascade impactor. The Geometric Standard Deviation (GSD) of the MMAD was calculated as 2.85.

The actual aerosol concentrations of 5.03 mg test item/L air was measured at the animals’ nose.

The mean actual exposure concentration of test item was as follows:

 

actual concentration	nominal concentration	mass median aerodynamic diameter	respirable amount particle size ≤4 µm
[mg/L air]	[mg/L air]	[µm]	[mg/L air]	[%]
5.03	27.78	2.585	2.00	39.7

 

Conclusion

Under the present test conditions, the LC₅₀value for rats following inhalation of test item for 4 hours was determined as follows (actual concentration, mean aerosol concentration):

LC₅₀ males and females combined (14 days) > 5.03 mg test item/L air/4 hours (actual concentration).

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 030 mg/m³

Quality of whole database:

The study is a guideline study with Klimisch score 1 (reliable without restrictions).

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

According to the EC Regulation 1272/2008 and subsequent regulations and based on the results of the acute oral and inhalation studies in rats the test substance must not be classified.