1-chloro-4-[(prop-2-yn-1-yloxy)methyl]benzene

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-02-24 to 2015-11-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Experimental study according to guideline and GLP

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 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

2008

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: U.S. EPA Health Effects Test Guidelines. OPPTS 870.3050; Jul 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany

Test material

Test material information

Constituent 1

Reference substance name:

1-chloro-4-[(prop-2-yn-1-yloxy)methyl]benzene

EC Number:

813-130-5

Cas Number:

4039-86-5

Molecular formula:

C10H9ClO

IUPAC Name:

1-chloro-4-[(prop-2-yn-1-yloxy)methyl]benzene

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services GmbH, Sulzfeld, Germany
- Age at study initiation: 42 ± 1 days
- Weight at study initiation: mean values fomr treatment groups: male 152.8-155.6, females 127.9-133.0
- Fasting period before study: no
- Housing: 5 animals per cage, in H-Temp polysulfonate cages type 2000P
- Diet: ad libitum, ground Kliba maintenance diet mouse/rat “GLP”, meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerland
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
The weighted test substance was mixed with food. Preperation was mixed once per week and stored in freezer for 4 days and 4 days at room temperature.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analyses of the test-substance preparations were carried out in compliance with GLP. The stability of the test substance in the diet in a freezer for a period of 4 days and, thereafter, for 4 days at room temperature was proven before the start of the administration period. Homogeneity was verified in 3 samples in the highest and lowest concentration (was used as a concentration control at the same time) at the beginning of the study; additional concentration control analyses were done in the mid concentration. The samples were taken from the specific food containers.

Duration of treatment / exposure:

4 weeks

Frequency of treatment:

continuoulsy

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0, 150, 500 and 1500 ppm
Basis:
nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
132 (male) and 130 (female) mg/kg bw/d (= 1500 ppm)
Basis:
actual ingested

Doses / concentrations 3

Remarks:

Doses / Concentrations:
41 (male) and 42 (female) mg/kg bw/d (= 500 ppm)
Basis:
actual ingested

Doses / concentrations 4

Remarks:

Doses / Concentrations:
13 (male) and 14 (female) mg/kg bw/d (= 150 ppm)
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: In a previously performed test study (Supporting study) the test substance was administered via the diet to groups of 5 male and 5 female Wistar rats for 3 or 14 days at concentrations of 0, 3000 and 10000 ppm. At 10000 ppm, a reduction of food and water consumption as well as body weight loss was observed between study days 0 and 3. All animals were sacrificed in a moribund condition on study day 3. At 3000 ppm, a reduction of food and water consumption was also observed as well as an initial body weight loss between study days 0 to 3. After 14 days, body weights were significantly lower by -28 % in males and -16 % in females compared to control animals. Several hematology and clinical chemistry parameters were affected and absolute as well as relative liver weights were increased in both sexes. Therefore, based on these results the concentrations 1500, 500 and 150 ppm in the diet were selected for the present study. The oral route was selected since this was proven to be suitable for the detection of a toxicological hazard.

Positive control:

not applicable

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: The animals were examined for evident signs of toxicity or mortality twice a day (in the morning and in the late afternoon) from Mondays to Fridays and once a day (in the morning) on Saturdays, Sundays and public holidays.
- Cage side observations checked in table No. 1 were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the administration period and thereafter at weekly intervals

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight was determined before the start of the administration period in order to randomize the animals. During the administration period the body weight was determined on day 0 (start of the administration period) and thereafter at weekly intervals.

FOOD CONSUMPTION AND COMPOUND INTAKE:
Food consumption was determined weekly over a period of 3 days and calculated as mean food consumption in grams per animal and day.

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
Water consumption was determined weekly over a period of 3 days and calculated as mean water consumption in grams per animal and day.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after administration period
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes, at least 16 hours
- How many animals: all
- Parameters checked in table No. 5 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after administration period
- Animals fasted: Yes, at least 16 hours
- How many animals: all
- Parameters checked in table No. 6 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: after administration period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, at least 16 hours
- Parameters checked in table No. 7 were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at the end of the administration period
- Dose groups that were examined: all
- Parameters checked in tables No. 2,3,4 were examined.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, see table No. 8
HISTOPATHOLOGY: Yes, see table No. 9

Statistics:

Means and standard deviations of each test group were calculated. In addition other analysis were performed for some parameters.
- Body weight + change: A comparison of each group with the control group was performed using DUNNETT's test (two-sided) for the hypothesis of equal means.
- rearing, grip strength forelimbs + hindlimbs, footsplay test, motor activity: Non-parametric one-way analysis using KRUSKALWALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON test (two-sided) for the equal medians.
- Blood parameters: For parameters with bidirectional changes: Non-parametric one-way analysis using KRUSKAL-WALLIS test. If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the hypothesis of equal medians. For parameters with unidirectional changes: Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) with Bonferroni-Holm adjustment for the hypothesis of equal medians.
- Urinalysis parameters (not pH, urine volume, specific gravity, color and turbidity): Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians.
- Urine pH, volume, specific gravity: Non-parametric one-way analysis using KRUSKAL-WALLIS test. If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the hypothesis of equal medians.
- Pathology weight parameters: Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting pvalue was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the equal medians.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No findings were observed in animals of any test group including the controls.

BODY WEIGHT AND WEIGHT GAIN
Mean body weights of male and female animals (1500 ppm) were lower compared to the control group, significantly on study day 28 (-11 %) in males and on study day 21 (-10 %) in females. Mean body weight change values of male and female animals in (1500 ppm) were significantly lower over the entire study period, showing a maximum of -32 % on study day 7 in males and -34 % on study day 21 in females. In the other treatement groups (150 and 500 ppm) no effects were detected.

FOOD CONSUMPTION AND COMPOUND INTAKE
During numerous time intervals, food consumption values for male and female animals of test group 1-3 (150, 500 and 1500 ppm) were slightly lower when compared to the control group. However, these values reflected the normal range of biological variation inherent in the strain of rats used for this study.

WATER CONSUMPTION AND COMPOUND INTAKE
No test substance-related, adverse changes with regard to water consumption were observed.

HAEMATOLOGY
No treatment-related changes among hematological parameters were observed.

CLINICAL CHEMISTRY
No treatment-related, adverse changes among clinical chemistry parameters were observed.

URINALYSIS
No treatment-related, adverse changes among urinalysis parameters were observed.

NEUROBEHAVIOUR
No test substance-related effects were observed.

ORGAN WEIGHTS
Abolute organ weights: Females treated with 1500 ppm had statistically significant changes in liver (126 % ) and females treated with 500 ppm in spleen (113 %).
Relative organ weights: Statistically significant changes were seen in kidneys for all females (150 ppm: 109 %, 500 ppm: 111 %, 1500 ppm: 115 %) and in liver for males at 1500 ppm (122 %) and females (500 ppm: 108 %, 1500 ppm: 138 %).

GROSS PATHOLOGY
All findings occurred individually. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

HISTOPATHOLOGY: NON-NEOPLASTIC
In the liver of male animals of test group 3 (1500 ppm) all animals revealed a minimal to slight increase of karyocytomegaly in the intermediate zone (between the periportal and centrilobular area). There was an increase in nuclear size and cell size of hepatocytes. The karyocytomegaly was
regarded to be treatment-related and as the meaning of this finding was not clear an adverse effect could not be excluded.
Females of the same test group revealed a minimal to slight centrilobular hypertrophy. These findings were regarded to be treatment-related but not adverse. All other findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

Effect levels

Dose descriptor:

NOAEL

Effect level:

41 - 42 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight, organ weights (liver); histopathology (liver) / equivalent to 500 ppm

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Results

 

Absolute organ weights

When compared to control group 0 (set to 100%), the mean absolute weights of following organs were significantly changed (statistically significant changes printed in bold):

	Male animals			Female animals
Test group (ppm)	1 (150)	2 (500)	3 (1500)	1 (150)	2 (500)	3 (1500)
Terminal body weight	100%	95%	88%*	96%	99%	92%
Liver				95%	107%	126%**
Spleen				101%	113%*	96%

*p ≤ 0.05; **p ≤ 0.01

 

Relative organ weight

When compared to control group 0 (set to 100%), the mean relative weights of following organs were significantly changed (statistically significant changes printed in bold):

	Male animals			Female animals
Test group (ppm)	1 (150)	2 (500)	3 (1500)	1 (150)	2 (500)	3 (1500)
Liver				109%**	111%**	115%**
Spleen	98%	99%	122%*	100%	108%*	138%**

*p ≤ 0.05; **p ≤ 0.01

 

Histopathology

Treatment-related findings were observed in the liver of male and females with incidences and grading according to the table below:

Liver	Male animals				Female animals
Test group (ppm)	0 (0)	1 (150)	2 (500)	3 (1500)	0 (0)	1 (150)	2 (500)	3 (1500)
No. of animals	5	5	5	5	5	5	5	5
Karyocytomegaly, intermediate	0	0	0	5
Grade 1				4
Grade 2				1
Karyocytomegaly, intermediate	0	0	0	0	0	0	0	5
Grade 1								4
Grade 2								1