Phosphoric acid, 2-ethylhexyl ester, ammonium salt

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 March 2017 to 24 October 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted in accordance with international guidelines and in accordance with GLP. All guideline validity criteria were met.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Ammonium mono(2- ethylhexyl)phosphate

Molecular formula:

C8H22NO4P

IUPAC Name:

Ammonium mono(2- ethylhexyl)phosphate

Constituent 2

Reference substance name:

Ammonium bis(2-ethylhexyl) phosphate

EC Number:

225-615-3

EC Name:

Ammonium bis(2-ethylhexyl) phosphate

Cas Number:

4971-47-5

Molecular formula:

C16H38NO4P

IUPAC Name:

Ammonium bis(2- ethylhexyl)phosphate

Constituent 3

Reference substance name:

2-ethylhexyl diphosphate ammonium salts / esters of 2-ethylhexan-1-ol with diphosphoric acid and their corresponding ammonium salts

Molecular formula:

C8H23NO7P2

IUPAC Name:

2-ethylhexyl diphosphate ammonium salts / esters of 2-ethylhexan-1-ol with diphosphoric acid and their corresponding ammonium salts

Test material form:

liquid

Details on test material:

Storage Conditions: Room temperature.

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd., Margate
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 8-9 weeks
- Weight at study initiation: All within ±20 % of the mean value
- Fasting period before study: Yes
- Housing: The animals were housed in groups of up to five during the acclimatisation period in suspended, solid floor cages with wire lids. From the day prior to dosing (Day-1), the rats were housed in groups of three in similar cages. Bedding was provided on a weekly basis to each cage by use of clean European soft wood bedding (Datesand Ltd., Manchester, UK). Each batch of bedding was analysed for specific constituents and contaminants. No contaminants were present in bedding at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Diet (e.g. ad libitum): Throughout the study the animals had access to 5LF2 EU Rodent Diet 14%, which was freely available to the animals at all times, except for a period of fasting from the evening of the day prior to dosing (Day-1) until approximately 3 hours after dosing. Each batch of diet had been analysed for specific constituents and contaminants by the manufacturer.
- Water (e.g. ad libitum): Mains water was provided ad libitum via cage mounted water bottles. The water was periodically analysed for specific contaminants.
- Acclimation period: The condition of the animals was assessed daily throughout the acclimatisation period of 7 to 10 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 ºC
- Humidity (%): 45-65 %
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: From: 14 March 2017 To: 27 April 2017

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 and 30 mg/mL for the 2000 and 300 mg/kg dose groups, respectively.
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: Not reported.
- Lot/batch no. (if required): n/a
- Purity: n/a

MAXIMUM DOSE VOLUME APPLIED: Dose volume applied at 10 mL/kg (actual volumes dispensed not reported)

DOSAGE PREPARATION (if unusual): n/a

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Data from the non-salt form of this test article indicated no deaths at dose levels of greater than 2000 mg/kg, therefore the first dose level for this study was 2000 mg/kg.

Doses:

2000 and 300 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical signs were recorded immediately post-dose, at approximately 15 and 30 minutes post-dose, hourly between 1 and 4 hours post-dose (inclusive), twice daily on Days 2, 3 and 4 and once daily from the fifth to last day of the observation period. Individual body weights were recorded on Day-1 (day before dosing) and on Days 1, 4, 8 and 15. Decedent carcass weights were also recorded prior to necropsy
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs and body weight.

Statistics:

n/a

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Three animals treated at 2000 mg/kg were found dead on Day 2. One other animal treated at this dose level was killed in extremis on this day.

No deaths were noted in animals treated at 300 mg/kg.

Clinical signs:

other: Mouth rubbing was noted immediately after dosing in three animals treated at 2000 mg/kg. Additional clinical signs noted in the animals that died during the study were piloerection, hunched posture, tremors, ptosis and lethargy. No clinical signs were se

Gross pathology:

Abnormalities noted at necropsy of one animal treated at 2000 mg/kg that died during the study were gaseous distension of the stomach, duodenum and jejunum and red mucosal surface of the cardia region of the stomach.

No abnormalities were noted at necropsy of all other animals that died or were killed in extremis during the study or at necropsy of those animals that were killed at the end of the study.

Any other information on results incl. tables

Table 2       Mortality Data

Dose (mg/kg bw)	Mortality (# dead/total)	Time range of deaths (day)
2000	4/6	2
300	0/6	n/a

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Based on the conditions of this study, the test article was classified as Category 4 in respect of its acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

Executive summary:

OECD 423 (2017) - In an acute oral toxicity study (OECD 423), groups of fasted, 8-9 week old, female Crl:WI(Han) rats were given a single oral dose of Reaction mass of Ammonium mono(2-ethylhexyl)phosphate, Ammonium bis(2-ethylhexyl)phosphate and 2-ethylhexyl diphosphate ammonium salts in water) at doses 2000 and 300 mg/kg bw and observed for 14 days.

Oral LD₅₀ Females = >300 - 2000 mg/kg bw.

According to the UN GHS classification, the test item can be classified as Category 4 in respect of its acute oral toxicity.