Phosphoric acid, 2-ethylhexyl ester, ammonium salt

Endpoint summary

Description of key information

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 at doses of 2000 and 300 mg/kg bw and observed for 14 days. Oral LD₅₀ (female) = >300 - 2000 mg/kg bw.

 

OECD 402 (2018) - In an acute dermal toxicity study (OECD 402), groups of 9 - 11 week old, female Crl:WI(Han) rats were given a single semi-occlusive dose of Reaction mass of Ammonium mono(2-ethylhexyl)phosphate, Ammonium bis(2-ethylhexyl)phosphate and 2-ethylhexyl diphosphate ammonium salts in water) at a dose of 2000 mg/kg bw and observed for 14 days. Dermal LD₅₀ (female) = >2000 mg/kg bw.

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:

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.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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

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

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

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.

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

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

> 300 - < 2 000 mg/kg bw

Quality of whole database:

A key study is submitted that was conducted in accordance with international guidelines and in accordance with GLP. The presented data are consistent with data requirements for the registrant’s tonnage band

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
In accordance with Annex VIII, Section 8.5.3, Column 2 of REACH, testing by the inhalation route does not need to be conducted if:
If exposure of humans via inhalation is unlikely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size.
The vapour pressure of the substance is 0.00018 Pa at 20 ºC. As per ECHA Guidance - Chapter R7.a (v6.0, July 2017), Section R.7.4.4.1.1; waiving of the inhalation route should be considered for substances with a vapour pressure of < 0.01 Pa. The vapour pressure of the substance falls below this cut off and therefore the inhalation route can be waived.

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 December 2017 to 21 May 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study performed under GLP. All relevant validity criteria were met.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

09 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Han Wistar obtained from Charles River (UK) Ltd., Margate

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: 9 - 11 weeks
- Weight at study initiation: All within ±20 % of the mean value
- Fasting period before study: No
- 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. 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: not stated

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: the day before treatment Electric clippers were used to remove all hair from the dorsum .
- % coverage: Approximately 10% of total body surface
- Type of wrap if used: A dense gauze patch was placed over the treated skin and retained in place by an elasticated, open-weave, adhesive compression bandage

REMOVAL OF TEST SUBSTANCE
- The dermal test site of each rat was lightly brushed clean of any solid residues and swabbed with water-moistened cotton wool before the animal was returned to the holding cage.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 2000 mg/kg bw
- Constant volume or concentration used: 1.049 g/mL

Duration of exposure:

24h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations and mortality checks were conducted at approximately 0.15, 0.5, 1, 2, 3, 4 hours and subsequently twice daily on Days 2, 3 and 4 and once daily from the fifth to last day of the observation period. Local effects were examined once daily for 14 days after the completion of the 24-hour exposure period. Full details on the scoring and criteria (consistent with Draize) are given in the full study report. Individual bodyweight were recorded prior to application of the test item on Day -1 (before dosing) and on Days 1, 4, 8 and 15.
- Necropsy of survivors performed: yes

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

There were no deaths

Clinical signs:

other: There were no clinical signs of reaction to treatment

Gross pathology:

No macroscopic changes were noted at necropsy.

Table  1. Dermal Reactions at Dose level 2000 mg/kg

Day	Dermal Reaction	Animal Number
		2	3	4
2	Erythema Oedema Other	2 0 -	2 0 -	2 0 -
3	Erythema Oedema Other	3 0 SC	2 0 SC	2 0 SC
4	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
5	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
6	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
7	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
8	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
9	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
10	Erythema Oedema Other	1 0 SC	2 0 SC	2 0 SC
11	Erythema Oedema Other	0 0 SC, FU	2 0 SC	2 0 SC
12	Erythema Oedema Other	0 0 SC, FU	2 0 SC	2 0 SC
13	Erythema Oedema Other	0 0 SC, FU	1 0 SC	1 0 SC
14	Erythema Oedema Other	0 0 SC, FU	1 0 SC	1 0 SC
15	Erythema Oedema Other	0 0 SC, FU	0 0 SC	0 0 SC

Key:

-       No other dermal changes apparent

SC       Scabbing

FU       Patchy hair growth

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in female Han Wistar (Crl:WI(Han)) strain rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Executive summary:

A study was performed according to OECD TG 402; Acute Toxicity (Dermal) and in accordance with GLP to assess the acute dermal toxicity of the test substance in the Han Wistar (Crl:WI(Han)) strain rat. Three animals were given a single dose of the test item, 24 hour under occluded dermal application of the undiluted test item to intact skin at a dose level of 2000 mg/kg body weight followed by 14 days observation.

There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. All animals showed expected gains in body weight during the study. There was well defined to moderate erythema was noted at the application sites of all treated animals on Days 2 and 3, with very slight to well-defined erythema persisting up to Day 14. Other adverse dermal reactions noted were scabbing and patchy hair growth which developed from Day 3 and persisted to the end of the observation period.

Under the conditions of this study, the dermal LD50 was established to exceed 2000 mg/kg bw in female Han Wistar (Crl:WI(Han)) strain rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

A key study is submitted that was conducted in accordance with international guidelines and in accordance with GLP. The presented data are consistent with data requirements for the registrant’s tonnage band

Justification for classification or non-classification

Based on the results of the acute oral toxicity study, conducted in accordance with OECD 423, the substance meets the classification criteria for Acute Tox. 4 (H302: Harmful if swallowed), in accordance with Regulation (EC) No 1272/2008 (CLP). No other criteria for acute toxicity were met.