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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Experimental data published in peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1978

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
Study not designed to meet a particular guideline.
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name of test material (as cited in study report): Diethyl phthalate
- Analytical purity: ester content min. 99%,

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River UK Ltd., Margate, Kent UK
- Age at study initiation: Not specified
- Weight at study initiation: Not specified
- Fasting period before study: N/A
- Housing: Not specified for standard short term study; singly housed for pair feeding study
- Diet (e.g. ad libitum): yes
- Water (e.g. ad libitum): yes
- Acclimation period: Not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 50-60%
- Air changes (per hr): Not Specified
- Photoperiod (hrs dark / hrs light): Not specified

Administration / exposure

Route of administration:
oral: feed
Vehicle:
other: Spratts laboratory diet No.1
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): Not specified
- Mixing appropriate amounts with (Type of food): Not specified
- Storage temperature of food: Not specified
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
N/A
Duration of treatment / exposure:
Two, 6 and 16 weeks for standard study
16 weeks for pair feeding study
Frequency of treatment:
continuously in the diet for 2, 6 and 16 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
0 (control), 0.2, 1.0 and 5.0 % for standard study and 0 (control) and 5% for pair feeding study a
Basis:
nominal in diet
No. of animals per sex per dose:
5 males and 5 females for 2 and 6 week phases
15 males and 15 females for 16 week phase
6 males and 6 females for pair feeding study
Control animals:
yes, plain diet
Details on study design:
Standard study: Groups of 15 rats of each sex were given diets containing 0 (control), 0.2, 1.0 or 5.0% DEP for 16 weeks.Additional groups of 5 rats of each sex were fed similar diets for 2 or 6 weeks. Body weight, food and water consumption consumption were measured at intervals. On completion of the designated treatment period the rats were deprived of food overnight and killed by exsanguination from the aorta under barbiturate anaesthesia. Blood samples were collected from the aorta for haematological and serum (at 16 weeks only) examination.
An autopsy was performed and macroscopic lesions were noted and selected organs weighed, sampled and the samples preserved in 10% buffered formalin. These tissues were processed to paraffin wax blocks, sectioned and stained with haematoxylin and eosin for histological examination.

Pair feeding study:
Groups of 6 rats of each sex, individually housed and fed untreated diet (controls) or diet containing 5% DEP for 16 weeks. Each control animal was from the same litter as one of the treated rats of the same sex. The treated rats were fed ad libitum and the weight consumed by each one was recorded daily. Each control rat was given a weight equal to that consumed by its paired litter mate in the previous 24 h period.Body weights were recorded at intervals during the treatment period.
Positive control:
N/A

Examinations

Observations and examinations performed and frequency:
Standard short term study:
Body weight, food and water intake at weekly intervals.
Blood samples for haematological and serum (after 16 weeks only) investigation and were taken at necropsy.
Urine samples were collected for examination during weeks 2, 6 and 13.

Pair feeding study:
Food consumed daily
Body weights weekly
Sacrifice and pathology:
The rats were killed by exanguination from the aorta under barbiturate anaesthesia. Macroscopic lesions were noted and the brain, pituitary, thyroid, heart, liver, kidney, adrenal glands, spleen, gonads, stomach, small intestine and caecum (with and without it's contents) were weighed. Samples of these organs and of salivary glands, trachea, lung, aorta, lymph nodes, yhymus, urinary bladder, oesophagus, colon, rectum, pancreas, uterus or prostate and seminal vesivcles, skeletal muscle, eye, hardarian gland and sciatic nerve were preserved in 10% buffered formalin. Parafin wax sections of the tissues were stained with haematoxylin and eosin for histological examination.
Other examinations:
Heamatology parameters examined after 2, 6 or 16 weeks included: haemoglobin concentration, packed cell volume, red blood cell and total white cell counts. Differential leucocyte counts were evaluated in control and 5.0% rats on blood fims made after 2, 6 and 16 weeks of treatment. In addition marrow smears were prepared and stained but not examined as there were no effects of treatment on the other haematological parameters examined.
The serum collected at week 16 was examined for effects on SGPT, SGOT and lactic dehydrogenase.
The urine collected during weeks 2, 6 and 16 of treatment was examined for the presence of cells and other microscopic constituents and semi quantitative tests were carried out for protein, glucose, ketones, bile and blood.
A concentration test was performed using the volume and SG of urine collected over a 6 h period of water deprivation.and in a 4h period commencing after 16 h without water.
In weeks 6 and 13 a dilution test was performed by bmaking the same measurements on urine produced in a 2 h period following a 25 ml/kg water load.
Statistics:
Student's t test for haematology, urine analysis, relative organ weights
Wilcoxon Rank Test for body weight and food consumption

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
Mortality not specifically mentioned
Mortality:
no mortality observed
Description (incidence):
Mortality not specifically mentioned
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Standard short term study: Throughout the treatment period both sexes given 5% DEP and female rats given 1% DEPin the diet gained significantly less weight than the controls. Transient lower body weight gains were seen in males given 1% DEP.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Overall food consumption was significantly lower than that of controls in rats of both sexes given 5% and females given 1%.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
Slight changes, some of which attained statistical but not biological significance.
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Description (incidence and severity):
Slight changes, some of which attained statistical but not biological significance.
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Increases in the absolute and relative weights of stomach and small intestine in rats of both sexes and in female rats given 0.2 and 1% DEP in the diet.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Abnormalities seen in testis of 1 male rat in 0.2 mgkg/day group.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
only abnormalities commonly seen in this strain of rat
Histopathological findings: neoplastic:
not examined
Details on results:
BODY WEIGHT AND WEIGHT GAIN
Throughout the treatment period both sexes given 5% DEP and female rats given 1% DEP in the diet gained significantly less weight than the controls. Transient lower body weight gains were seen in males given 1% DEP.

FOOD CONSUMPTION AND COMPOUND INTAKE
Overall food consumption was significantly lower than that of controls in rats of both sexes given 5% and females given 1%. Total group mean intakes of DEP were 150, 770 & 3160 and 150, 750 & 3170 mg/kg/day in treated males and females respectively.

WATER CONSUMPTION AND COMPOUND INTAKE
No effects
HAEMATOLOGY
After 6 weeks of treatment the erthrocyte count in male rats given 5% DEP in the diet was significantly higher than in their control counterparts. This change was associated with a non significant increase in haemoglobin levels when compared with the relevant controls.

CLINICAL CHEMISTRY
No effects

URINALYSIS
Male rats given 5% DEP excreted significantly fewer cells than controls in week 13. After a 6 hour water deprivation period in Weeks 2 & 6 female& male rats given 5% DEP respectively produced more urine of a slightly higher SG than the controls, though this effect was not seen after the 20 hour period of deprivation.
In the 2 hour dilution test in week 6 male & female rats given 5% DEP produced significantly larger volumes of more dilute urine & smaller volumes of less dilute urine than controls. In week 13 the urine of male rats given 5% DEP had a significantly higher SG.

ORGAN WEIGHTS
In rats of both sexes given 5% DEP in the diet for 16 weeks the absolute weights of the brain, heart, spleen & kidneys were statistically significantly lower than controls. There were also reductions in the absolute weights of the gonads (females) & heart and the spleen & kidneys of males given 5% DEP in the diet after 2/6 weeks of treatment. The weights of stomach of rats given DEP at 5% in the diet increased at times during the study but attained statistical significance with respect to controls after 2 (males) and 2 & 16 (females) weeks of treatment. The weight of the full caecum was also increased in females given 5% DEP in the diet for 16 weeks.
In the 1% DEP dietary group after 2 weeks of treatment, increased kidney weights were seen in males & lower pituitary weights in females. Increased gonad weights were also seen in females after 6 weeks of treatment at the 1% dietary level.
The relative weights of the brain, kidney (week 16 only) liver, stomach, small intestine and full caecum were increased in rats of both sexes & of testes of males given 5% DEP in the diet with respect to the controls. In the rats given 0.2 or 1.0% there were similar increases in the relative weights of the liver, stomach & small intestine.

GROSS PATHOLOGY
Abnormalities seen in testis of 1 male rat in 0.2 mgkg/day group but no histological changes were seen.

HISTOPATHOLOGY: NON-NEOPLASTIC
Some fatty degeneration and slight vacuolation of the liver, pyleonephritis & lymphocytic infiltration of the kidney were observed. Incidence not dose related and all common findings of this strain of rat.

OTHER FINDINGS
None reported


Pair feeding study
CLINICAL SIGNS AND MORTALITY
Not reported
BODY WEIGHT AND WEIGHT GAIN
Rats given 5% DEP in the diet lost more weight than their respective controlsover the 1st day of treatment. Weight gain in the 5% group was also lower than in controls there after. The difference attained statistical significance in week 16.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
The total food consumed by the rats given 5% DEP in the diet was greater than that consumed by their respective controls.
FOOD EFFICIENCY
Not reported
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)
Not reported


Effect levels

Dose descriptor:
NOAEL
Effect level:
150 mg/kg diet
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Effects on body weight and organ weight in 1.0 (750 mg/kg) and 5.0% (3160 mg/kg) dose groups

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion