Reaction mass of C12-14 tert-alkylamines and dimethyl hydrogen phosphate and methyl dihydrogen phosphate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 January 2018 to 10 May 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NouSan No. 8147, 24 November 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of C12-14 tert-alkylamines and dimethyl hydrogen phosphate and methyl dihydrogen phosphate

EC Number:

948-071-5

IUPAC Name:

Reaction mass of C12-14 tert-alkylamines and dimethyl hydrogen phosphate and methyl dihydrogen phosphate

Test material form:

liquid

Specific details on test material used for the study:

- Purity: >98% (nominal); This substance has a Unknown or Variable composition, is a Complex reaction product, or a Biological material (UVCB)
- Description: Yellow liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 8 to 12 weeks of age
- Weight at study initiation: Ranged from 170 to 179 g
- Fasting period before study: Yes, overnight immediately before dosing
- Housing: Suspended solid floor polypropylene cages with stainless steel mesh lids and furnished with softwood flake bedding (Datesand LTD., Cheshire, UK).
- Diet (e.g. ad libitum): Free access to 2014C Teklad Global Certified Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK
- Water (e.g. ad libitum): Free access to tap drinking water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19 to 25 °C
- Humidity: 30 to 70% respectively
- Air changes: At least fifteen changes per hour
- Photoperiod: 12 hours continuous light and 12 hours darkness

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

- The test item was freshly prepared, as required, as a solution in dimethyl sulfoxide.
- Dimethyl sulfoxide was used as it produced the most suitable formulation at the required concentration.
- The test item was formulated with no correction for the test item purity within 2 hours of being applied to the test system. It was assumed that the formulation was stable for this duration.
- No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. (This is an exception with regard to GLP and has been reflected in the GLP compliance statement within the study report).

Doses:

2000 and 300 mg/kg

No. of animals per sex per dose:

One animal per dose, followed by five animals per dose

Control animals:

no

Details on study design:

Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose and a single animal was treated with this dose.

Due to mortality at a dose level of 2000 mg/kg, an additional animal was treated at a dose level of 300 mg/kg. In the absence of toxicity at a dose level of 300 mg/kg, an additional group of animals was treated at this dose level. A total of five animals were therefore treated at a dose level of 300 mg/kg in the study.

All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. At least 24 hours was allowed between each dose group to confirm the survival of the previously dosed animals.

Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.

Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.

At the end of the observation period the surviving animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

300 mg/kg bw

Based on:

test mat.

Mortality:

At the dose level of 2000 mg/kg, the (one) animal was found dead thirty minutes after dosing.
At the dose level of 300 mg/kg, there were no unscheduled deaths.

Clinical signs:

At the dose level of 2000 mg/kg, no signs of systemic toxicity were noted during the observation period.
At the dose level of 300 mg/kg, hunched posture was noted in one animal during the day of dosing. No other signs of systemic toxicity were noted.

Body weight:

Dose level of 2000 mg/kg - see tables section, below.
At the dose level of 300mg/kg, based on historical data from the supplier for this strain all animals showed expected gains in body weight over the observation period.

Gross pathology:

At the dose level of 2000 mg/kg, abnormalities noted at necropsy were dark liver, dark kidneys and reddened gastric mucosa.
At the dose level of 300 mg/kg, no abnormalities were noted at necropsy.

Any other information on results incl. tables

Individual Necropsy Findings -2000 mg/kg

Dose Level (mg/kg)	Animal Number and Sex	Time of Death	Macroscopic Observations
2000	1-0 Female	Found dead 30 minutes after dosing	Liver: Dark Kidneys: Dark Gastric Mucosa: Reddened

 

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 to 2000 mg/kg body weight and classed as Category 4, according to GHS.

Executive summary:

The study was performed to the standardized guideline OECD 420, under GLP conditions to assess the acute oral toxicity of the test item in the Wistar strain rat.

 

Following a sighting test at dose levels of 2000 mg/kg and 300 mg/kg body weight, an additional four fasted females was given a single oral dose of the test item, as a solution in dimethyl sulfoxide, at a dose level of 300 mg/kg body weight followed with a 14-day observation period. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

The animal treated at a dose level of 2000 mg/kg was found dead thirty minutes after dosing and all animals dosed at 300 mg/kg survived to the scheduled sacrifices. Hunched posture was noted in one animal treated at a dose level of 300 mg/kg during the day of dosing. No other signs of systemic toxicity were noted. Based on historical data from the supplier for this strain, surviving animals showed expected gains in body weight over the observation period. Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg that was found dead during the study were dark liver, dark kidneys and reddened gastric mucosa.

 

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 to 2000 mg/kg body weight and classed as Category 4, according to GHS.