Hexa-1,5-diene

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental start date: 10 January 2018. Experimental completion date: 02 February 2018.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines

Version / remarks:

JMAFF Guidelines (2000), including the most recent revisions

Deviations:

no

GLP compliance:

yes

Test type:

up-and-down procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hexa-1,5-diene

EC Number:

209-754-7

EC Name:

Hexa-1,5-diene

Cas Number:

592-42-7

Molecular formula:

C6H10

IUPAC Name:

hexa-1,5-diene

Specific details on test material used for the study:

1,5-hexadiene. 99.4% purity.

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BSC-433-4-0488-5
- Expiration date of the lot/batch: 01 June 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature protected from light

Analyses conducted to support the information cited in the certificate of analysis for the test item were not conducted in compliance with the GLP or GMP regulations. The characterization of the test item was conducted under a sponsor or sponsor subcontractor quality system.

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adult animals (approximately 12 weeks old)
- Weight at study initiation: 172 to 211 g
- Fasting period before study: overnight prior to dosing and until 3-4 hours after administration of the test item
- Housing: On arrival and following assignment to the study, animals were individually housed (pilot and full study) or group housed (limit study, up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room(s) in which the animals were kept were documented in the study records. Animals were separated during designated procedures/activities. Each cage was clearly labeled.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water: Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period: The animals were allowed to acclimate to the Test Facility for at least 5 days before the commencement of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 21ºC
- Humidity (%): 44 to 51%
- Air changes (per hr): 10 or greater
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

IN-LIFE DATES: 16 January to 02 February 2018

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The Test Item, 1,5-hexadiene, was administered as received. Adjustment was made for specific gravity of the test item. No correction was made for the purity/composition of the test item. The dose volume for each animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows:
Dose level (g/kg) / spec.gravity or density (g/mL).
The dosing formulations were stirred continuously during dose administration. Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the test item. Water was available.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

The toxicity of the test item was assessed in a limit test by treatment of five females at a dose level of 2000 mg/kg body weight.
The toxicity of the test item was assessed by stepwise treatment of five females. The first animal was treated at a dose level of 2000 mg/kg. Based on the absence of mortality, four more females were dosed at 2000 mg/kg. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dosages.
A single dose of test item was administered to the appropriate animals by oral gavage on Day 1, using a syringe with a plastic gavage cannula attached.
Postdose observations. Postdose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days. All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate). Signs were graded for severity and the maximum grade was predefined at 3 or 4. Grades were coded as slight (grade 1), moderate (grade 2), severe (grade 3) and very severe (grade 4). For certain signs, only its presence (grade 1) or absence (grade 0) was scored.
Body weights. Animals were weighed individually on Day 1 (predose), 8 and 15. A fasted weight was recorded on the day of dosing.
Terminal procedures: All moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

Not required

Results and discussion

Preliminary study:

Not applicable

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

other: Hunched posture, uncoordinated movements, piloerection and/or salivation were noted for the animals on Day 1.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

No other findings reported.

Any other information on results incl. tables

Clinical signs, test day 1, females 2000 mg/kg:

Test day	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
Hours after treatment	0	2	4
Hunched posture (Max. Grade: 1)
Animal 1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 2	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 3	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 4	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 5	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Piloerection (Max. Grade: 1)
Animal 1	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 2	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 3	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 4	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 5	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Salivation (Max. Grade: 3)
Animal 1	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 2	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 3	N/A	N/A	N/A	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 4	N/A	N/A	N/A	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 5	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-

 

Body weights (gram):

Day	1	8	15
Animal 1	187	208	213
Animal 2	211	250	252
Animal 3	172	189	198
Animal 4	183	208	218
Animal 5	181	207	209
Mean (n=4, animals 2 to 5)	187	214	219
SD (n=4, animals 2 to 5)	17	26	23

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The oral LD50 value of 1,5-hexadiene in Wistar rats was established to exceed 2000 mg/kg body weight.
Based on these results, 1,5-hexadiene does not have to be classified for acute oral toxicity according to CLP Regulation.

Executive summary:

The objective of this study was to determine the potential toxicity of 1,5-hexadiene, when given by oral gavage at a single dose to rats of a single sex to evaluate the potential reversibility of any findings.

Initially, 1,5-hexadiene was administered by oral gavage to one female Wistar rat at 2000 mg/kg body weight. Based on the absence of mortality, four additional female Wistar rats were dosed at 2000 mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred.

Hunched posture, uncoordinated movements, piloerection and/or salivation were noted for the animals on Day 1.

The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

No abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50 value of 1,5-hexadiene in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, 1,5-hexadiene is not classified for acute oral toxicity according to the CLP Regulation.