1,2-Benzenedicarboxylic acid, di-C16-18-alkyl esters

Repeated dose toxicity: oral

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

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study 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

Test material information

Constituent 1

Reference substance name:

1,2-Benzenedicarboxylic acid, mixed cetyl and stearyl esters

EC Number:

270-487-4

EC Name:

1,2-Benzenedicarboxylic acid, mixed cetyl and stearyl esters

Cas Number:

68442-70-6

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 or test system 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 / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0, 500, 1000 and 10000 ppm
Basis:
nominal in diet

Doses / concentrations 2

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

Doses / concentrations 3

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

1

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;

2

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;

3

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'

4

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.