Reaction mass of (E)-3-methyl-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one and (E) -1-(2,6,6-trimethylcyclohex-2-en-1-yl)pent-1-en-3-one and (E) -1-(2,6,6-trimethylcyclohex-1-en-1-yl)pent-1-en-3-one

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline study with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

yes

Remarks:

BASF AG, Department of Toxicology

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ionone, methyl-

EC Number:

215-635-0

EC Name:

Ionone, methyl-

Cas Number:

1335-46-2

Molecular formula:

C14H22O

IUPAC Name:

Ionone, methyl

Details on test material:

- Name of test material (as cited in study report): Methylionone
- batch number: 09049610046
- physical state/appearance: liquid, yellowish clear
- storage conditions: refrigerator, exclusion of oxygen (store under nitrogen)
- analysis of test substance: the test substance has been characterized by analysis; the stability of the test substance over the study period has been proven by reanalysis; the test substance was monogeneous by visual inspection
- analysis of test substance preparations: the stability of the test substance in olive oil DAB 10 for a time period of 4 hours was confirmed by analysis
- conduct of analytics: central analytical department of BASF AG

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- age: young adult
- weights at start of study: 150-300 g (+/- 20% of mean weight)
- identification: individual identification using cage cards and group identification by tail marking
- housing: single housing in fully air-conditioned rooms, central air conditioning guaranteed a range of 20-24°C for temperature and of 30-70% for relative humidity. There were no deviations from these ranges which influenced the results of the study.
- day/night rhythm: 12:12
- type of cage: stainless steel wire mesh cages, type DK-III
- bedding: no bedding in the cages, sawdust in the waste trays
- drinking water: tap water ad libitum per day
- diet: Kliba-Labordiät 343, Klingentalmühle AG, Kaiseraugst, Switzerland, ad libitum
- analysis of drinking water: the drinking water is regularly assayed for chemical contaminants by municipal authorities of Frankenthal and the technical services of BASF AG as well as for the presence of germs by a contract laboratory
- analysis of feed: the feed used in the study was assayed for chemical and microbiological contaminants
- acclimatization period: acclimatization for at least 1 week
- fasting period: the animals were given no feed at least 16 hours before administration, but water was available ad libitum

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: olive oil DAB 10

Details on oral exposure:

- reason for the vehicle: test substance could not be homogeneously distributed in aqua bidest
- form of administration: solution
- amounts administered: dose: 2000 mg/kg; concentration: 40 g/100 ml; administration volume: 5 ml/kg
- time of day of administration: in the morning

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- observation period: 17 days for males / 14 days for females
- body weight determination: individual body weights shortly before application (day 0), weekly thereafter and at the end of the study (before fasting period)
- signs and symptoms: recording of signs and symptoms several times on the day of administration and at least once each workday for the indiviual animals
- mortality: a check for any dead or moribund animals was made twice each workday and once on saturdays, sundays and on public holidays
- pathology: necropsy at the last day of the observation period; withdrawal of food at least 16 hours before killing with CO2; then necropsy with grosspathology examination; necropsy of all animals that died before end of study as early as possible

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

male/female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: no deaths observed

Mortality:

no mortality up to day 14 (females) and day 17 (males)

Clinical signs:

see table

Body weight:

see table

Gross pathology:

necropsy findings of sacrificed animals: organs without particular findings (3 males, 3 females)

Other findings:

clinical symptoms reversible

Any other information on results incl. tables

animal symptoms:

	males		females
	cageside observations	number of animals	cageside observations	number of animals
impaired general state	H1 - H4	3	H0 - H2	2
poor general state			H0 - D2	3
dyspnoea	H1 - H4	3	H0 - D2	3
apathy			H0 - D2	3
staggering	H1 - H4	3	H0 - D2	3
twitching			H4	1
saltatory spasm	H1 - H3	1
spastic gait			D1 - D2	1
piloerection			H0 - D2	3
smeared fur			D2	1
exophthalmos			H0 - H1	3
S5			D2	1

H: hour

D: day

S5: red smeared fur in the anogenital area

body weights:

		day
		0	7	13
male	1	175	244	273
	2	175	230	263
	3	175	230	281
	mean	175	235	272
female	1	170	202	210
	2	171	194	215
	3	172	203	215
	mean	171	200	213

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The oral acute toxicity of methylionone was tested in a study under GLP performed according to OECD Guideline 423 (BASF, 1999). Three male and three female Wistar rats were once administered by gavage with 2000 mg/kg bw in 5 ml/kg bw olive oil DAB 10. The following observation time was 17 days for males and 14 days for females. Although signs of toxicity were noted during the first days after application, the animals appeared normal after five days. Since no mortality was observed, the LD50 cut-off value was determined to be 5000 mg/kg bw.