6-chlorohexan-2-one

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 10th to march 8th, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

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

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

6-chlorohexan-2-one

EC Number:

233-546-5

EC Name:

6-chlorohexan-2-one

Cas Number:

10226-30-9

Molecular formula:

C6H11ClO

IUPAC Name:

6-chlorohexan-2-one

Test material form:

liquid

Test animals

Species:

rat

Strain:

Wistar

Remarks:

(Cmdb: WI; outbred)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Husbandry of laboratory animals of the Experimental Medicine Centre at the Medical University in Białystok.
- Age and weight at study initiation: One 9-week-old animal weighing 233.0 g (dose of 300 mg/kg bw), and one 10-week-old animal weighing 220.0 g (dose of 2000 mg/kg bw) were used in the preliminary experiment. Four 11-week-old animals whose average body weight was 228.5 g (dose of 2000 mg/kg b.w.) were used in the main experiment.
- Fasting period before study: 19 hours
- Housing: The animals were kept in plastic cages covered with wire bar lids. The dimensions of the cages were 58 x 37 x 21 cm. In the preliminary experiment, the animals were caged individually. In the main experiment, there were four animals in one cage. UV-sterilized wood shavings were used as bedding. The environment of the animals was enriched by placing wooden blocks and nesting materials for laboratory animals in the cages.
- Diet (e.g. ad libitum): Murigran standard granulated laboratory fodder ad libitum.
- Water (e.g. ad libitum): tap water ad libitum.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 20-23ºC
- Humidity: 27-50%
- Air changes: 16 times/hour
- Photoperiod: 12 hours light/12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 60mg/mL (300 mg/kg bw) and 400 mg/mL (2000 mg/kg bw)
- Amount of vehicle (if gavage): 0.5 mL/100 g bw
- Justification for choice of vehicle: Low water solubility.
- Other: The suspensions were prepared on the administration day (shortly before the administration).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The starting dose for the sighting study was selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg bw as a dose expected to produce evident toxicity. Since no data were available, the sighting study commenced with the administration of the test item at a dose of 300 mg/kg bw to one female rat. Since no evident toxicity was produced, the test item at a single dose of 2000 mg/kg bw was administered to the second animal. On the grounds of the sighting study results (no evidence toxicity), the dose of 2000 mg/kg bw was selected to be used in the main study.

Doses:

Sighting study: 300 and 2000 mg/kg bw
Main study: 2000 mg/kg bw

No. of animals per sex per dose:

1 female for the preliminary experiment at 300 mg/kg bw.
5 females (included the one from preliminary) for the main experiment at 2000 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: General condition of the animals at least once a day, clinical detailed observations on day 0: 10, 30 and 60 minutes after the administration and then at hourly intervals up to 5th hour after administration. Thereafter once a day from the day 1 to the day 14.
Weight was determined on day 0 (directly before administration),7 and 14 before euthanasia.
- Necropsy of survivors performed: yes

Results and discussion

Preliminary study:

No evident toxicity was observed in dose 300 mg/kg bw and the animal survived the experiment. Following single administration of the test item at a dose of 2000 mg/kg bw, the following changes were observed: the rounded back, wavering gait, slightly, then distinct decreased locomotor activity, animal could be very east to catch, and dejection and bristled coat. The animal survived the experiment.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortalities were recorded, all animals survived the experiment.

Clinical signs:

On the day of administration of the test item, rounded back, wavering gait, and slightly decreased locomotor activity was observed.

Body weight:

Body weight gain was observed in all animals. See table 1 on "Any other information on results incl. tables"

Gross pathology:

Gross examinations did not reveal any pathological changes in the examined animals.

Any other information on results incl. tables

Table 1. Body weight of the animals (g) overall list

Dose (mg/kg body weight)	Animal number	Body weight (g)				Body weight gain (g) 0 to 14
		Day of the experiment
		0	7		14
300	1*	233	267	279		46
2000	1*	220	242	259		39
	2	233	260	266		33
	3	225	227	237		10
	4	221	247	255		34
	5	235	251	262		27

*Female from the preliminary experiment.

Table 2. Summary of results.

Dose of test item (mg/kg b.w.)	300	2000
	Preliminary experiment	Preliminary experiment	Main experiment
Mortality	0/1	0/1	0/4
Clinical signs	Not stated	rounded back, wavering gait, slight and then distinct decreases of locomotor activity, animal was very easy to catch, dejection, bristled coat.	In all animals: rounded back, wavering gait, slight decreases of locomotor activity.

Applicant's summary and conclusion

Interpretation of results:

other: Not classified (CLP Regulation EC no. 1272/2008)

Conclusions:

The oral-LD50 for the test substance in female rats was greater than 2000 mg/kg bw.

Executive summary:

An acute toxicity study was conducted with the test item according to the fixed dose procedure specified in OECD guideline 420 and EU Method B.1.bis, under GLP conditions. Since no data was available, the preliminary experiment started with the administration of the test item suspended in corn oil at a dose of 300 mg/kg b.w. to one animal. Since no evident toxicity was observed at 300 mg/kg bw, the test item at a single dose of 2000 mg/kg b.w. was administered to the second animal. On the grounds of the preliminary experiment results, the dose of 2000 mg/kg bw was selected to be used in the main experiment and it was applied to four further rats. The observation period consisted of 14 days and included detailed clinical observations, body weight gain records and gross examinations at termination. In the main experiment (2000 mg/kg bw) changes in the body posture, the gait, and the locomotor activity were observed in all animals. The animals survived the experiment. Gross examinations did not reveal any pathological changes in the examined animals. Under the conditions of this study, the acute oral LD50 was determined to be greater than 2000 mg/kg bw in female rats