Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental Phase: 16 August 2018 to 02 October 2018.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Updated information. Study now available and provided ahead of extended submission date of 31/05/2019.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

There was a deviation from the study plan where mean humidity dropped below the minimium target level on two days. This deviation to the study plan is not considered to have any impact on the integrity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

There was a deviation from the study plan where mean humidity dropped below the minimum target level on two days. This deviation to the study plan is not considered to have any impact on the integrity of the study.

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

There was a deviation from the study plan where mean humidity dropped below the minimium target level on two days. This deviation to the study plan is not considered to have any impact on the integrity of the study.

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

EC Number:

271-672-2

EC Name:

Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

Cas Number:

68603-74-7

Molecular formula:

The substance is a UVCB so there is no molecular formula

IUPAC Name:

Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

Test material form:

solid

Test animals

Species:

rat

Strain:

other: Wistar Han

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: approximately 8-12 weeks old
- Weight at study initiation: 136 to 221 g
- Housing: Animals were group housed (up to 3 animals of same dosing group together) in polycarbonate cages containing sterilized sawdust as bedding material. Animals were separated during designated procedures/activities.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): Relative humidity of 37 to 69%
- Air changes (per hr): Ten or greater air changes per hour with 100% fresh air (no air recirculation)
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 1% Aqueous carboxymethyl cellulose for 2000 mg/kg (200 mg/mL) and propylene glycol for the dose level of 300 mg/kg (30 mg/mL),

Details on oral exposure:

VEHICLE
1% Aqueous carboxymethyl cellulose was initially selected as the vehicle for formulations at 2000 mg/kg (200 mg/mL). For the dose level of 300 mg/kg (30 mg/L) formulations were observed to not be homogenous. Based on additional trial formulations, propylene glycol was selected as suitable vehicle at this dose level.

- Concentration in vehicle: 2000 mg/kg = 200 mg/L, 200 mg/kg = 30 mg/L
- Amount of vehicle (if gavage): The dose volume for each animal was based on the body weight measurement prior to dosing. A dose volume of 10 mL/kg body weight was used for each dose.
- Justification for choice of vehicle: Based on trial preparations performed at the Test Facility to select the suitable vehicle and to establish a suitable formulation procedure.

MAXIMUM DOSE VOLUME APPLIED: of 10 mL/kg body weight

Doses:

The toxicity of the test item was assessed by stepwise treatment of groups of 3 females. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines.

The first group was treated at a dose level of 2000 mg/kg. Based on the results, two additional groups were dosed at 300 mg/kg.

No. of animals per sex per dose:

Three animals at 2000 mg/kg
Six animals at 300 mg/kg (dose as two groups of three)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Post dose observations were made at least three times on the day of dosing and once daily thereafter. Animals were weighed on Day 1 (predose) and on days 8 and 15. Terminal body weights were collected from animals found dead or euthanized moribund after Day 1.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs

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:

There were three unscheduled deaths on the study. At 2000 mg/kg, one animal was found dead on Day 8. One animal was killed in extremis on Day 8 and another on day 10.

Clinical signs:

At 2000 mg/kg, lethargy, hunched posture, piloerection, yellow discoloration of the genital region, chromodacryorrhoea (nose), diarrhoea, lean appearance and/or red discolouration of the mouth and nose were noted for the animals between Days 1 and 10. At 300 mg/kg, Lethargy, hunched posture, uncoordinated movements, shallow respiration, piloerection and/or ptosis were noted for the animals between Days 1 and 8.

Body weight:

The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

Gross pathology:

Abnormalities of the adrenal glands (enlarged, both sides), thymus (reduced in size) and forestomach (irregular surface) were found in the animals that were sacrificed for humane reasons during the study, at macroscopic post mortem examination. Macroscopic post mortem examination of the surviving animals at termination did not reveal any abnormalities.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The oral LD50 value of Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates in Wistar Han rats was established to be within the range of 300-2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

Executive summary:

Introduction

The study was conducted todetermine the oral acute toxicity of Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates, to the rat. The study was designed to meet the requirements of the following guidelines:

·        OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method"

·        EC No 440/2008, part B: "Acute Oral Toxicity, Acute Toxic Class Method"

·        EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

·        JMAFF Guidelines (2000), including the most recent revisions.

 

Method

Initially, the test item was administered by oral gavage to three female Wistar Han rats at 2000 mg/kg body weight. In a stepwise procedure, two additional groups of three females were dosed at 300 mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

Results

At 2000 mg/kg, one animal was found dead on Day 8. One animal was killed in extremis on Day 8 and another on day 10.  At 300 mg/kg, no mortality occurred.

At 2000 mg/kg, lethargy, hunched posture, piloerection, yellow discoloration of the genital region, chromodacryorrhoea (nose), diarrhoea, lean appearance and/or red discoloration of the mouth and nose were noted for the animals between Days 1 and 10. At 300 mg/kg, Lethargy, hunched posture, uncoordinated movements, shallow respiration, piloerection and/or ptosis were noted for the animals between Days 1 and 8.

The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

Abnormalities of the adrenal glands (enlarged, both sides), thymus (reduced in size) and forestomach (irregular surface) were found in the animals that were sacrificed for humane reasons during the study, at macroscopic post mortem examination. Macroscopic post mortem examination of the surviving animals at termination did not reveal any abnormalities.

Conclusion

The oral LD50 value of Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates in Wistar Han rats was established to be within the range of 300-2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

According to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.