Phosphoric acid, mono- and di-C12-14-alkyl esters

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29th August 2019 - 1st October 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Adopted 17 December 2001.

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

30 May 2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

17 December 2001

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, Acute oral toxicity (2-1-1), 12 Nousan No 8147, Agricultural Production Bureau.

Version / remarks:

November 24, 2000.

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Water

Constituent 2

Reference substance name:

C12-C14 alcohol

IUPAC Name:

C12-C14 alcohol

Constituent 3

Reference substance name:

Mono (C12-C14 alkyl) phosphate ester

IUPAC Name:

Mono (C12-C14 alkyl) phosphate ester

Constituent 4

Reference substance name:

Di (C12-C14 alkyl) diphosphate ester

IUPAC Name:

Di (C12-C14 alkyl) diphosphate ester

Constituent 5

Reference substance name:

Tri (C12-C14 alkyl) triphosphate ester

IUPAC Name:

Tri (C12-C14 alkyl) triphosphate ester

Constituent 6

Reference substance name:

C12-C14 alkyl pyrophosphate esters

IUPAC Name:

C12-C14 alkyl pyrophosphate esters

Constituent 7

Reference substance name:

Ortho-phosphoric acid

IUPAC Name:

Ortho-phosphoric acid

Constituent 8

Reference substance name:

unknown

Molecular formula:

unknown

IUPAC Name:

unknown

Test material form:

solid: bulk

Specific details on test material used for the study:

Appearance: Solid, whitish
Storage conditions: Room temperature (10-30°C) in the dark
Supplier: Sponsor
Batch number: 1023S17201
Expiry date: 09 July 2020
Purity: 100%

Test animals

Species:

rat

Strain:

other: WIST albino

Remarks:

RccHan™

Sex:

female

Details on test animals or test system and environmental conditions:

Animal information.
Healthy nulliparous and non-pregnant female RccHan™:WIST albino rats were obtained from Envigo RMS (UK) Ltd.
The animals were allocated without conscious bias to cages within the treatment groups. They were housed in groups of one or four rats of the same sex.
Each animal was identified uniquely within the study by tail marking. Each cage label was color-coded and was identified uniquely with the study number, dose level and animal mark.
The animals were allowed to acclimatize to the conditions described below for at least five days before treatment. For those animals selected for this study, their body weights were in the range 159 to 175 g and they were approximately eight to twelve weeks of age prior to dosing (Day 1). The body weight variation did not exceed +/- 20% of the mean body weight of any previously treated animals.

Animal care and husbandry.
Animals were housed inside a limited access rodent facility. The facility was designed and operated to minimize the entry of external biological and chemical agents and to minimize the transference of such agents between rooms.
The animal room was kept at positive pressure with respect to the outside by its own supply of filtered fresh air, which was passed to atmosphere and not re-circulated. The temperature and relative humidity controls were set to maintain the range of 20 to 24 deg.C and 40 to 70%, respectively. Any minor deviations from these ranges would not have had an adverse effect on the animals and would not affect the integrity or validity of the study. Artificial lighting was controlled to give a cycle of 12 hours continuous light and 12 hours continuous dark per 24 hours. Environmental parameters are archived with the departmental raw data.
Periodic checks were made on the number of air changes in the animal rooms. Temperature and humidity were monitored daily.
Alarms were activated if there was any failure of the ventilation system, or temperature limits were exceeded. A stand-by electricity supply was available to be automatically brought into operation should the public supply fail.
The cages were solid bottomed polycarbonate cages with a stainless steel mesh lid. Each cage contained a quantity of autoclaved softwood bark-free fiber bedding. Cages, food hoppers, water bottles and bedding were changed at appropriate intervals.
The animals were allowed free access to a standard rodent diet (Teklad 2014C Diet), except for overnight prior to and approximately four hours after dosing. This diet contained no added antibiotic or other chemotherapeutic or prophylactic agent.
Potable water taken from the public supply was freely available via polycarbonate bottles fitted with sipper tubes.
Each cage of animals was provided with Aspen chew blocks or balls for environmental enrichment. Chew blocks or balls were provided throughout the study and were replaced when necessary. Each cage of animals was provided with a plastic shelter for environmental enrichment, which was replaced at the same time as the cages.
Each batch of diet was analyzed routinely by the supplier for various nutritional components and chemical and microbiological contaminants. Supplier’s analytical certificates were scrutinized and approved before any batch of diet was released for use. The quality of the water supply is governed by regulations published by the Department for Environment, Food and Rural Affairs. Certificates of analysis were received routinely from the water supplier.
No other specific contaminants that were likely to have been present in the diet or water were analyzed, as none that may have interfered with or prejudiced the outcome of the study was known.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

In the absence of data regarding the toxicity of the substance, 300 mg/kg was chosen as the starting dose.
In the preliminary study: 1 animal/ dose administered 300 mg/kg (as 30 mg/ml of corn oil, dose volume 10 mL/kg)
1 animal/ dose administered 2000 mg/kg (as 200 mg/ml of corn oil, dose volume 10 mL/kg).
In the absence of mortality or toxicity at a dose level of 2000 mg/kg in the preliminary test, an additional group of 4 animals treated at 2000 mg/kg (as 200 mg/kg of corn oil, dose volume 10 mL/kg).

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

For 300 mg/kg dose: 1 female rat in the preliminary study.
For 2000 mg/kg dose: five female rats treated in total (1 female rat in preliminary study and 4 female rats in the main study).

Control animals:

no

Details on study design:

Dose Administration.
The appropriate dose volume of the substance was administered to each rat by oral gavage using a plastic syringe and plastic catheter.
A record of the weight of each formulation dispensed and the amount remaining after dosing was made. The balance of these two weights was compared with the predicted usage as a check that the doses had been administered correctly.
Formulations were stirred before and throughout the dosing procedure.

Serial Observations.

1) Mortality: Cages of rats were checked at least twice daily for any mortalities.

2) Clinical Observations: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1. On subsequent days, animals were observed once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only). The nature and severity, where appropriate, of any clinical signs and the time were recorded at each observation. All animals were observed for 14 days after dosing.

3) Body Weight: The weight of each rat was recorded on Days -1, (prior to dosing), 8 and 15. Individual weekly body weight changes and group mean body weights were calculated.

Terminal Investigations.
Method to kill:
All animals were humanely killed on Day 15 by carbon dioxide asphyxiation and subsequently exsanguinated.

Macroscopic Pathology:
All animals were subject to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities. The macroscopic appearance of the brain, cecum, duodenum, heart, kidneys, small and large intestine, liver, lungs and bronchi, spleen, stomach, subcutaneous tissue and urinary bladder was recorded.

Statistics:

N/A

Results and discussion

Preliminary study:

Mortality: No deaths reported during the study.

Clinical signs: Piloerection was observed on Day 1 for female number 2 dosed at 2000 mg/kg. This sign was seen approximately 4 hours after dosing and recovery, as judged by external appearance and behavior, was complete by 5 hours after dosing.

Body weight: All animals were considered to have achieved satisfactory body weight gains throughout the study.

Macroscopic examination: Macroscopic examination at study termination on Day 15 revealed a yellow liquid present in the stomach, duodenum, small and large intestines in female number 1 dosed at 300 mg/kg.

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50

Effect level:

300 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

Macroscopic examination on day 15 revealed yellow liquid present in the stomach, duodenum, small and large intestines.

Effect levels 2

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 during the study.

Clinical signs:

other: Piloerection was observed on Day 1 for female number 2 dosed in the preliminary study at 2000 mg/kg. This sign was seen approximately 4 hours after dosing and recovery, as judged by external appearance and behavior, was complete by 5 hours after dosing.

Gross pathology:

Macroscopic examination at study termination on Day 15 revealed a yellow liquid present in the stomach, duodenum, small and large intestines in female number 1 dosed in the preliminary study at 300 mg/kg. No abnormalities were revealed in any other animal at the macroscopic examination at this time.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

LD50> 2000 mg/kg bw

Conclusions:

The LD50 of the substance is considered to be greater than 2000 mg/kg body weight after single oral administration to female WIST albino (RccHan) rats.

Executive summary:

In a reliable in vivo acute oral toxicity study conducted in accordance to OECD guideline 420, the substance was administered to female RccHan^TM WIST albino rats in corn oil (vehicle) as a single dose via oral gavage route. In preliminary study female rats administered 300 mg/kg bw (one animal) and 2000 mg/kg bw (one animal) dose. No deaths or signs of toxicity have been observed at any dose level. Therefore, further 4 female rats were administered the substance at 2000 mg/kg bw. All animals survived in the highest dose group of 2000 mg/kg body weight. The substance did not cause death or evident signs of toxicity. During the observation period of 15 days, no other signs of intoxication, change of health, nor any other adverse reactions were seen. Macroscopic examination of the animals at the end of the study noted that one animal dosed at 300 mg/kg bw had yellow liquid present in the stomach, duodenum, small and large intestines. No other abnormalities were revealed in any other animal at the macroscopic examination. The LD50 of the substance is considered to be greater than 2000 mg/kg bw after single oral administration to female WIST albino (RccHan) rats.