Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
21 May 1971 - 06 Jan 1972
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (no rationale for dose selection given (middle dose level is 1/10 of highest dose level))

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1972
Report Date:
1972

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
: lack of details on test substance; as animals were housed in groups of 5 per cage the calculation of compound intake was not individually, but calculated as mean values from each cage; No rationale for dose selection given (middle dose level is 1/10 of
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): ester of long chain fatty alcohol C16-18 esterified with phthalic acid used as a slip additive
- Physical state: no data
- Analytical purity: no data
- Batch: no identification on received test compound given
- Other: Date of arrival: March 18th, 1971

Test animals

Species:
rat
Strain:
other: CFY strain
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Carworth Europe, Alconbury, Huntingdon
- Housing: 5 per cage with wire-mesh floor.
- Diet (ad libitum): Spiller's Laboratory Small Animals Diet
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 50 ± 5

IN-LIFE DATES: From: May, 21st, 1971 continued for 13 weeks.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was dispersed in corn-oil heated to 40ºC, and then incorporated into the diet.

DIET PREPARATION
A premix containing 20000 ppm and 4 % corn oil was prepared each week, and from this various dietary concentrations were obtained by direct dilution with further quantities of diet.
Corn oil was added so that all diets, including control diet, contained 2% corn oil. Homogeneity was achieved by mixing for 10 minutes in a rotary double-cone blender. The diets were then stored until use in heatsealed, opaque polythene bags.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
continuously with diet
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0, 500, 1000 and 10000 ppm
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
0, 34, 68 and 683 mg/kg bw/day
Basis:
other: group mean intake (males) as calculated from group mean bodyweight and food consumption
Remarks:
Doses / Concentrations:
0, 39, 75 and 772 mg/kg bw/day
Basis:
other: group mean intake (females) as calculated from group mean bodyweight and food consumption
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Positive control:
No

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
The quantity of food consumed by each cage (not by each animal) was recorded weekly.

FOOD EFFICIENCY:
The efficiency with which food was utilized was assessed by calculation of mean food conversion ratios (FCR values) during the period of most rapid growth, the ratio representing the weight of food consumed per unit gain in bodyweight.


WATER CONSUMPTION was assessed by inspection of the water bottles. Regular measurement of water intake was not introduced since there was no evidence of a treatment-related effect.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Before treatment, 6 and 12 weeks after treatment commenced.
- Dose groups that were examined: Animals of the vehicle control and highest dose groups.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at weeks 4, 8 and 12 from 10 male and 10 female animals from control and highest dose groups.
After 12 weeks, also 10 males from th 1000 ppm - group were analyzed for red cell parameters.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- Parameters checked: PCV, Hb, RBC, MCV, MCHC, PCV, Differential count,

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at weeks 4 and 12 from 5 male and 5 female animals of the control and highest dose group.
After 12 weeks additional animals from the 1000 ppm group were analyzed for AP and GPT. After 13 weeks, males of the 1000 ppm group were examined for serum proteins.
- Animals fasted: No data
- Parameters checked: Urea, Glucose, Total serum proteins, AP, GPT, GOT, Sodium, Potassium

URINALYSIS: Yes
- Time schedule for collection of urine: weeks 4, 8 and 12 from 5 male and 5 female animals of control and highest dose groups.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked: pH, specific gravity, protein, reducing substances, glucose, ketones, bile pigments, urobilin, blood pigments, urine sediment


NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes, adrenals, brain, colon, duodenum, eyes, heart, ileum, kidneys, liver, lungs, lymph nodes, ovaries, pancreas, pituitary, spleen, stomach, testes, thymus, thyroids, urinary bladder, uterus of animals of control and highest dose groups. Additionally pancreas of rats from the 1000 ppm group were analyzed.
Other examinations:
ORGAN WEIGHTS of adrenals, brain, heart, kidneys, liver, ovaries, pituitary, spleen, testes, thyroids, uterus.
For inter-group comparison, relative organ weights were calculated as percentages of bodyweight.
Statistics:
Student's t-test

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
two male rats of the highest dose died
Mortality:
mortality observed, treatment-related
Description (incidence):
two male rats of the highest dose died
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
decreased body weight gain in the highest dose group
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
The overall food intake of males of the highest dose group was slightly less than that of the controls.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
at highest dose
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
effects observed, treatment-related
Description (incidence and severity):
in one male of the highest dose group
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
non-adverse effects
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Increased levels of serum AP in both sexes of the highest dose group.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
slightly increased thyroid weights in males of the highest dose group
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
internal haemorrhages at highest dose males
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Haemorrhage and associated chronic inflammatory reaction in the pancreas and areas of fat, throughout the body in male rats of the highest dose group.
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
At 10000 ppm two deaths were observed among male rats.
Between weeks 6 and 10 only, slight pallor of the extremities was noted in several male rats of the highest dose group.
No other reaction to treatment was observed.

BODY WEIGHT AND WEIGHT GAIN
At 10000 ppm slight depression in rate of bodyweight gain in males and to a lesser extent in females was observed. No other significant differences were observed between control and treated group.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
The overall food intake of males of the highest dose group was slightly less than that of the controls. No other significant differences between control and treated groups were observed.

FOOD EFFICIENCY
Slightly decreased efficiency was seen in rats receiving 10000 ppm during the first week. This did not, however, affect the overall performance, this being comparable with that of the controls.
No decrease in efficiency was observed in the remaining treated groups.

WATER CONSUMPTION
no effects

OPHTHALMOSCOPIC EXAMINATION
In one male rat of the highest dose group extensive haemorrhage was seen in both eyes in the posterior chamber. This change was considered in relation to the changes seen during autopsy.

HAEMATOLOGY
no adverse effects were detected.

CLINICAL CHEMISTRY
Serum AP levels were abnormally high in both males and females of the highest dose group during weeks 4 and 12. Extension of the investigation to rats receiving 1000 ppm during week 12 revealed a normal value, although it was noted that the value in a single male rat was abnormally high.
The mean levels of GPT were also higher in males of the highest dose group compared to controls, but the individual values were well within the normal range, as also were the slightly depressed serum protein concentrations found in these animals during week 12. Animals of the 1000 ppm group showed values comparable to controls.
The only adverse reaction was considered to be the elevated serum AP levels in male and female rats receiving 10000 ppm in the diet.

URINALYSIS
no effects

ORGAN WEIGHTS
Slightly increased thyroid weights in males of the highest dose group.
Thyroids of male animals of the highest dose group were statistically significantly different from the control value.

GROSS PATHOLOGY
Internal haemorrhage in both male animals of the highest dose group, which died during the study period.
Change compatible with the occurence of haemorrhage were seen in all but one of the male rats that had received the highest dose in the diet.

HISTOPATHOLOGY: NON-NEOPLASTIC
Haemorrhage and associated chronic inflammatory reaction in the pancreas and in areas of fat throughout the body in male rats of the highest dose group. No other treatment-related histopathological changes were found.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
1 000 ppm
Based on:
test mat.
Remarks:
equivalent to approx. 68 mg/kg bw/d
Sex:
male
Basis for effect level:
other: No abnormalities based on clinical signs; mortality; body weight; food consumption; food efficiency; ophthalmoscopic examination; haematology; clinical chemistry; urinalysis; gross pathology; organ weights; histopathology;
Dose descriptor:
NOAEL
Effect level:
1 000 ppm
Based on:
test mat.
Remarks:
equivalent to approx. 75 mg/kg bw/d
Sex:
female
Basis for effect level:
other: No abnormalities based on clinical signs; mortality; body weight; food consumption; food efficiency; ophthalmoscopic examination; haematology; clinical chemistry; urinalysis; gross pathology; organ weights; histopathology;
Dose descriptor:
LOAEL
Effect level:
10 000 ppm
Based on:
test mat.
Remarks:
equivalent to approx. 683 mg/kg bw/d
Sex:
male
Basis for effect level:
other: see 'Remark'
Dose descriptor:
LOAEL
Effect level:
10 000 ppm
Based on:
test mat.
Remarks:
equivalent to approx. 772 mg/kg bw/d
Sex:
female
Basis for effect level:
other: Based on increased levels of serum AP

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Blood chemistry - group mean values

Group

Week No.

Urea mg%

Total Glucose mg%

SAP

KA units

GPT

SF units

GOT SF units

Electrolytes, mEq/l

Na+

K+

male control animals

4

32

113

89

39

159

142

4.7

male

10000 ppm

4

37

112

140**

51**

119

144

5.0

female control animals

4

43

128

86

34

148

141

4.3

female

10000 ppm

4

38

122

116*

37

136

139

4.5

male control animals

12

40

124

59

32

121

143

5.1

male

1000 ppm

12

 -

-

63

36

 -

 -

male

10000 ppm

12

42

128

98**

54***

96*

144

4.5

female control animals

12

51

121

54

45

128

142

4.8

female

1000 ppm

12

 -

 -

52

39

 -

 -

 -

female

10000 ppm

12

41

121

93***

48

128

143

5.2

 

   *P < 0.05 (when compared with control value)

  **P < 0.01 (when compared with control value)

***P < 0.001 (when compared with control value)

 

Dietary administration at 500 and 1000 ppm was without overt adverse effects in all treated animals.

 

Applicant's summary and conclusion

Conclusions:
It can be assumed, that the subchronic NOAEL for the test substance could have been higher than the values identified in this study - under the aspect, that the middle dose level was only one tenth of the highest dose level.