Reaction mass of ethyl (2S,3S)-3,4-dibromo-2-methylbutanoate and ethyl (2R,3R)-3,4-dibromo-2-methylbutanoate and ethyl (2S,3R)-3,4-dibromo-2-methylbutanoate and ethyl (2R,3S)-3,4-dibromo-2-methylbutanoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-08-03 to 2009-10-01; day of dosing: 2009-08-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001-12-17

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008-05-30

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nosan No. 8147 as this in line with OECD 423

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:

ethyl (2R,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2R,3S)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3S)-3,4-dibromo-2-methylbutanoate

EC Number:

700-225-0

Cas Number:

1160806-44-9

Molecular formula:

C7 H12 Br2 O2

IUPAC Name:

ethyl (2R,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2R,3S)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3S)-3,4-dibromo-2-methylbutanoate

Details on test material:

- Name of test material (as cited in study report): 2-Methyl-3,4-dibrombuttersäureethylester
- Physical state: liquid/ colorless, clear
- Analytical purity: 98.1 wt%
- Lot/batch No.: 35684-12-5
- Stability under test conditions: The stability under storage conditions over the study period was guaranteed by BASF SE
- Storage condition of test material: Refrigerator; under light exclusion
- Other: density: 1.640 g/ml (determined by Bioassay)
No further data.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
female Wistar/Crl:WI (Han) rats
Rats were selected since this rodent species is recommended in the respective test guidelines. Wistar rats were selected since there is extensive experience available in the laboratory with this strain of rats. As suggested by the OECD guideline nulliparous and non-pregnant female animals were used for the test, because there is no indication that male animals are likely to be more sensitive to the acute effects of the test substance.
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: ca. 10 weeks
- Weight at study initiation: group mean body weights were in the range between 177 and 190 g.
- Fasting period before study: yes; feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: single housing in Makrolon cage, type III
- Diet (ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany)
- Water (ad libitum): tap water
- Acclimation period: at least 5 days before administration

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24°C
- Humidity (%): 20 - 80%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2009-07-31 To: 2009-09-02

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: unchanged (no vehicle) or olive oil

Details on oral exposure:

For the high dose, the liquid test substance was administered unchanged. For the low dose, the test substance was dissolved in olive oil.

1) 2000 mg/kg dose level
VEHICLE : no vehicle
MAXIMUM DOSE VOLUME APPLIED: 1.22 ml/kg

2) 50 mg/kg dose level
VEHICLE: Olive oil Ph.Eur.
- Concentration in vehicle: 4.1 g/100 ml; i.e. 4.1% (w/v)
- Amount of vehicle (if gavage): 1.22 ml/kg bw
- Justification for choice of vehicle: solubility of the test substance
Because no analyses of the test substance preparation were requested by BASF SE, no samples have been taken.
MAXIMUM DOSE VOLUME APPLIED: 1.22 ml/kg bw
DOSAGE PREPARATION (if unusual): The test-item preparation of the 50 mg/kg bw was produced for each test group shortly before administration by stirring with a magnetic stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
By the request of BASF SE, a starting dose of 50 mg/kg bw was chosen in the first step with 3 female animals. As no animal died, 2000 mg/kg bw was administered to 3 female rats in the second step. Because no animal died at the second step, 2000 mg/kg bw was administered to another group of 3 female animals in the third step. As no animal died in this step the study was terminated.

Doses:

2000 mg/kg bw, undiluted
50 mg/kg bw, as a 4.1% (w/v) solution in olive oil

No. of animals per sex per dose:

6 females for the 2000 mg/kg dose level (2 subgroups of 3)
3 females for the 50 mg/kg dose level

Control animals:

no

Details on study design:

Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
-- Individual body weights were determined shortly before administration (day 0), weekly thereafter and on the last day of observation.
-- Recording of signs and symptoms was made several times on the day of administration, and at least once daily thereafter each workday for the individual animals.
-- A check for any dead or moribund animal was made at least once each workdays.
- Necropsy of survivors performed: yes
- Other: No histological examinations were performed.

Statistics:

Calculations were performed using Microsoft Excel 2003 and checked with a calculator.

Results and discussion

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: No deaths occurred. Clinical symptoms were noted at 2000 mg/kg.

Effect levels 2

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: The combined acute oral LD50 was calculated.

Mortality:

No mortality occurred.

Clinical signs:

other: 2000 mg/kg bw groups: Clinical observation in two animals of the 2000 mg/kg bw test groups revealed impaired general state, dyspnea, piloerection, staggering and reduced feces from hour 3 until study day 2 after the administration. In one animal dosed w

Gross pathology:

There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period.

Any other information on results incl. tables

Table: individual and group mean body weights

Dose [mg/kg bw]	2000				2000				50
Animal no. Animal code	1 R130	2 R131	3 R132	mean ± SD	4 R148	5 R149	6 R150	mean ± SD	7 R108	8 R109	9 R110	mean ± SD
Body weight [g] at study day
0	190	185	195	190.0± 5.00	176	177	178	177.0± 1.00	178	183	179	180.0± 2.65
7	198	190	198	195.3± 4.62	182	167	190	179.7 ± 11.68	194	200	195	196.3± 3.21
14	215	203	209	209.0 ± 6.00	189	180	201	190.0± 10.54	207	208	202	205.7± 3.21

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Remarks:

Migrated information

Conclusions:

Under the conditions of this study the median lethal dose of 2-Methyl-3,4-dibrombuttersäureethylester after oral administration was found to be greater than 2000 mg/kg bw in rats.