[2R-(2α,3Z,5α)]-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, compound with tert-butylamine (1:1)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study performed according to method equivalent to OECD and EU Guidlines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

Safepharm Standard Method Number SPL 59

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

Safepharm Standard Method Number SPL 59

Principles of method if other than guideline:

Study performed according to Safepharm Standard Method Number SPL 59.

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

TBA Clavulanate

IUPAC Name:

TBA Clavulanate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): BRL 14151C
- Substance type: pure substance
- Physical state: cream-coloured powder
- Storage condition of test material: room temperature over silica gel
Container : clear glass jar
Date recieved : 14 August 1991

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: supplied by Charles River (UK) Ltd., Manston, Kent.
- Age at study initiation: 6-9 weeks
- Weight at study initiation: Male: 149-172, Female: 143-151
- Fasting period before study: overnight fast
- Housing: solid-floor polypropylene cages with sawdust bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: minimum acclimation period of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23°C
- Humidity (%): 40-69%
- Air changes (per hr): 15changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: suspension in 0.5% gum tragacanth

Details on oral exposure:

VEHICLE
- Concentration in vehicle: suspension in 0.5% gum tragacanth
- Amount of vehicle (if gavage): as stated above

MAXIMUM DOSE VOLUME APPLIED: no data (however 2000mg/kg @ 172g = 358mg)

Doses:

2000 mg/kg and 200 mg/kg

No. of animals per sex per dose:

3M+3F @ 2000 mg/kg and 1M + 1F @ 200 mg/kg

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed for overt signs of systemic toxicity 1/2, 1, 2 and 4 hours after dosing and then once daily for 14 days. Individual bodyweights were recorded on Day 0 (the day of dosing), on Day 7 and Day 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:Any Macroscopic findings were recorded and the affected tissues preserved in fixative (10% neutral buffered formaldehyde). The carcases and tissues showing no macroscopic lesions were also maintained in fixative.

Statistics:

Evaluation of data
From the Mortality data obtained, an estimate of the acute oral median lethal dose (LD50) and maximum non-lethal dose level was made, the estimated LD50 was used to classify the test material as follows:
Approximate LD50 > 2000 EEC: Unclassified
Approximate LD50 >200 to 2000 EEC: Harmful
Approximate LD50 >25 to 200 EEC: Toxic
Approximate LD50 <25 EEC: Very Toxic
The clinical observations and necropsy findings were examined and if possible the maximum no-observed effect level was identified.

Results and discussion

Effect levels

Effect levels 1

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

male/female

Dose descriptor:

other: NOEL

Effect level:

200 mg/kg bw

Based on:

test mat.

Mortality:

For mortality data, see Table 1 (below).

Clinical signs:

other: Individual clinical observations are given in Table 2 (below). lethargy was noted in one male and two femals two or four hours after dosing. Ptosis was also noted in one female two hours after dosing. No Clinical signs of toxicity were noted in animals tr

Gross pathology:

Individual necropsy findings are given in Table 4 (below).
No macroscopic abnormalities were noted at necropsy.

Any other information on results incl. tables

Table 1: Mortality Data:

Dose Level mg/kg	Sex	Number of Animals Treated	Deaths
2000	M	3	0/3
	F	3	0/3
200	M	1	0/1
	F	1	0/1

Table 2: Individual Clinical Observations and Mortality Data

Dose Level mg/kg	Animal Number & Sex	Effects Noted after Dosing (hours)				Effects Noted During Period After Dosing (Days)
		1	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	1-0 M	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-0 M	0	0	L	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-1 M	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-0 F	0	0	0	L	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	4-0 F	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	4-1 F	0	0	L Pt	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
200	5-0 M	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	6-0 F	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Where L = lethargy,                     Pt = ptosis,                0 = no signs of systemic toxicity

Table 3: Individual Bodyweights and Weekly Bodyweight Increases

Dose level mg/kg	Animal Number & Sex	Bodyweight (g) at Day			Increment (g) During Week
		0	7	14	1	2
2000	1-0  M	149	220	276	71	56
	3-0  M	152	214	273	62	59
	3-1  M	166	239	305	73	66
	2-0  F	149	195	210	46	15
	4-0  F	151	187	215	36	28
	4-1  F	151	199	233	48	34
200	5-0  M	172	265	334	93	69
	6-0  F	143	201	227	58	26

Table 4: Individual Necropsy Findings

Dose Level mg/kg	Animal Number & Sex	Macroscopic Observations
2000	1-0  M	No abnormalities detected
	3-0  M	No abnormalities detected
	3-1  M	No abnormalities detected
	2-0  F	No abnormalities detected
	4-0  F	No abnormalities detected
200	4-1  F	No abnormalities detected
	5-0  M	No abnormalities detected
	6-0  F	No abnormalities detected

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The approximate LD50 was found to be greater than 2000 mg/kg bodyweight. the test mateial was therefore classified as non-toxic if swallowed.
The no-observed effect level was found to be 200 mg/kg bodyweight.

Executive summary:

This study was performed to assess the acute oral lethality of the test material in the rat and to identify an approximate no-observed effect level. The results may be used as a basis for classification.

The acute oral toxicity of the test material was assessed by treatment of a group of fasted animals (three males and three females) at a dose level of a group of fasted animals (three males and three females) at a dose level of 2000 mg/kg bodyweight. The test material was administered as a suspension in 0.5% gum tragacanth.

There were no deaths. Lethargy was noted in three animals two or four hours after dosing.

All animals showed expected gain in bodyweight during the study.

No macroscopic abnormalities were noted at necropsy.

The no-observed effect level was determined by treatment of a pair of animals (one male and one female) at a lower dose level of 200 mg/kg bodyweight.

There were no deaths or clinical signs of toxicity. No macroscopic abnormalities were noted at necropsy.

The approximate LD50 was found to be greater tha 2000 mg/kg bodyweight.

The no-observed effect dose level was found to be 200 mg/kg bodyweight.

The test material was therefore classified as non-toxic if swallowed, according to 67/548/EEC Dangerous Substances Directive (DSD) and the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP)