N,N-dibutylbutan-1-aminium bis[2-(hydroxy-kO)benzoato(2-)-kO]borate(1-)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April to May 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline (OECD 423) compliant study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2014

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 (Acute Toxicity (Oral))

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:

4,4'-dioxo-4H,4'H-2,2'-spirobi[[1,3,2]benzodioxaborinin]-2-uide; tributylazanium

EC Number:

700-906-2

Cas Number:

22450-96-0

Molecular formula:

C26H36BNO6

IUPAC Name:

4,4'-dioxo-4H,4'H-2,2'-spirobi[[1,3,2]benzodioxaborinin]-2-uide; tributylazanium

Constituent 2

Reference substance name:

SABoTBA

IUPAC Name:

SABoTBA

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): SABoTBA
- Purity: 98.77%
- Purity test date: 08 April 2014
- Lot/batch No.: 12342
- Expiration date of the lot/batch: Stable under recommended storage conditions
- Storage condition of test material: Room temperature in the dark

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd.
- Age at study initiation: 8-9 weeks
- Weight at study initiation: 230-270 g
- Fasting period before study: Overnight prior to dosing and approximately four hours after dosing
- Housing: The cages were solid bottomed polycarbonate cages with a stainless steel mesh lid. Each cage contained a quantity of autoclaved softwood bark-free fibre bedding. Cages, food hoppers, water bottles and bedding were changed at appropriate intervals.
- Diet (e.g. ad libitum): Rat and Mouse No. 1 Maintenance Diet was available ad libitum, with the exception stated above
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): target 19-23°C
- Humidity (%): 40 to 70%
- Air changes (per hr): not reported, however periodic checks performed
- Photoperiod (hrs dark / hrs light): 12 h light/12 h dark

IN-LIFE DATES: From: 09 April 2013 To: 03 May 2013

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 1% methylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: A vehicle trial performed in 1% methylcellulose resulted in a white suspension that was suitable for dosing.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The dose levels for the study were chosen in compliance with the study guidelines. As no previous toxicological information was available the initial dose was 300 mg/kg.

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

2 groups of 3 females at each dose level (animals R1-R3 and R4-R6 at 300 mg/kg and R7-R9 and R10-R12 at 2000 mg/kg)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations twice daily and body weights weekly or at death
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs

Results and discussion

Preliminary study:

Not applicable

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 - <= 5 000 mg/kg bw

Based on:

test mat.

Mortality:

One female (R9) dosed at 2000 mg/kg died on Day 1. Clinical signs prior to death comprised fast breathing, flat posture, tremors, convulsion, partially or fully closed eyelids, irregular breathing, underactive behaviour, hindlimbs splayed, increased body tone, shallow breathing, salivation and reduced body temperature. These signs were seen from approximately 20 minutes after dosing. A loss in bodyweight was noted for the decedent. Macroscopic examination of the animal revealed congestion (characterised by darkened tissues/organs) of the subcutaneous tissue, liver and small and large intestines, pallor of the lungs and white liquid fluid contents in the stomach.
One female (R10) dosed at 2000 mg/kg died on Day 2. Clinical signs prior to death comprised fast breathing, hunched posture, tremors, piloerection, partially closed eyelids, irregular breathing, underactive behaviour, reduced body tone, flat posture, salivation and reduced body temperature. These signs were seen from approximately 20 minutes after dosing. A loss in bodyweight was noted for the decedent. Macroscopic examination of the animal revealed congestion (characterised by darkened tissues/organs) of the subcutaneous tissue, lungs and small and large intestines, a small caecum and dark red liquid fluid contents in the small intestines and yellow liquid fluid content in the large intestines.

Clinical signs:

other: Clinical signs of reaction to treatment comprised fast breathing, tremors, piloerection, hunched posture and unsteady gait seen in all females dosed at 2000 mg/kg. Underactive behaviour, irregular breathing, and reduced body temperature were seen in three

Gross pathology:

Macroscopic examination at study termination on Day 15 revealed a small stomach for one female (R8) treated at 2000 mg/kg. No abnormalities were revealed in any other animal at the macroscopic examination at this time.

Any other information on results incl. tables

Table 1 Mortality Data

Dose (mg/kg)	No. of deaths in groups of 3	Day
		1		2		3 to 15
		<5m	ca 4.5 h	a	b	a	b
300	0/3F	-	-	-	-	-	-
300	0/3F	-	-	-	-	-	-
2000	1/3F	0	1	0	0	0	0
2000	1/3F	0	0	1	0	0	0

a = first observation, b = second observation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute median lethal oral dose (LD50) to rats of SABoTBA was demonstrated to be between 2000 and 5000 mg/kg bodyweight.

Executive summary:

The study was performed to assess the acute oral toxicity of SABoTBA to the rat using the acute toxic class method (OECD Guideline 423). Two groups of three fasted female rats received a single oral gavage dose of the test substance, formulated in 1% methylcellulose, at a dose level of 300 mg/kg bodyweight. As results at this dose level indicated the acute (median) lethal oral dose of the test substance to be greater than 300 mg/kg bodyweight, in compliance with the study guidelines a further two groups of three fasted females were similarly dosed at 2000 mg/kg bodyweight to complete the study.

One female dosed at 2000 mg/kg died on Day 1. Clinical signs prior to death comprised fast breathing, flat posture, tremors, convulsion, partially or fully closed eyelids, irregular breathing, underactive behaviour, hindlimbs splayed, increased body tone, shallow breathing, salivation and reduced body temperature. These signs were seen from approximately 20 minutes after dosing. A loss in bodyweight was noted for the decedent. Macroscopic examination of the animal revealed congestion (characterised by darkened tissues/organs) of the subcutaneous tissue, liver and small and large intestines, pallor of the lungs and white liquid fluid contents in the stomach. One female dosed at 2000 mg/kg died on Day 2. Clinical signs prior to death comprised fast breathing, hunched posture, tremors, piloerection, partially closed eyelids, irregular breathing, underactive behaviour, reduced body tone, flat posture, salivation and reduced body temperature. These signs were seen from approximately 20 minutes after dosing. A loss in bodyweight was noted for the decedent. Macroscopic examination of the animal revealed congestion (characterised by darkened tissues/organs) of the subcutaneous tissue, lungs and small and large intestines, a small caecum and dark red liquid fluid contents in the small intestines and yellow liquid fluid content in the large intestines. Clinical signs of reaction to treatment in surviving animals comprised fast breathing, tremors, piloerection, hunched posture and unsteady gait seen in all females dosed at 2000 mg/kg. Underactive behaviour, irregular breathing, and reduced body temperature were seen in three females dosed at 2000 mg/kg. Partially closed eyelids, reduced body tone, flat posture and thin build were seen in two females dosed at 2000 mg/kg and lachrymation, brown staining on the head, convulsion and increased body tone were observed for individual animals dosed at 2000 mg/kg. These signs were first noted approximately 20 minutes after dosing. Recovery of surviving animals, as judged by external appearance and behaviour, was complete by Day 10. No clinical signs were seen in any animal dosed at 300 mg/kg. A low bodyweight gain was noted for two females receiving 300 mg/kg and two females receiving 2000 mg/kg on Day 15 and a bodyweight loss was recorded for one female receiving 300 mg/kg on Day 15. All other surviving animals were considered to have achieved satisfactory bodyweight gains throughout the study. Macroscopic examination at study termination on Day 15 revealed a small stomach for one female treated at 2000 mg/kg. No abnormalities were revealed in any other animal at the macroscopic examination at this time.

The acute median lethal oral dose (LD50) to rats of SABoTBA was demonstrated to be between 2000 and 5000 mg/kg bodyweight.