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Description of key information

Oral subacute and subchronic repeated dose toxicity of the reaction mass of Stearoylbenzoylmethane and Palmitoylbenzoylmethane were assessed in a 28-day rat study and a 90-day dog study, respectively. In the absence of relevant toxicity at any dose levels in both studies, the NOAEL was therefore the highest dose tested i.e 10000 mg/kg/day in the 28-day rat study and 1350 mg/kg/day in the 90-day dog study.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
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: Study conducted before GLP principles (1977). limited information on test conditions and no data on purity of test substance. However, the study is conducted according to scientific principles of guidelines
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 409 (Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents)
Deviations:
not specified
GLP compliance:
no
Remarks:
Study before GLP guidelines
Limit test:
no
Species:
dog
Strain:
Beagle
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS:
- Source: no data
- Age at study initiation: 10-13 months
- Weight at study initiation: no data
- Fasting period before study: no data
- Housing: two animals were housed per cage
- Food consumption: U.A.R. N°103 dry food given in meal form, ad libitum
- Water consumption: no data
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS:
- Temperature: no data
- Humidity: no data
- Air changes: no data
- Photoperiod: no data

- IN-LIFE DATES: no data
:
Route of administration:
oral: feed
Vehicle:
not specified
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: no data


DIET PREPARATION
- Rate of preparation of diet (frequency): test substance dietary concentrations in each diet mix were prepared each 2 weeks
- Mixing appropriate amounts with (Type of food): no data
- Storage temperature of food: no data


VEHICLE: no data

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
No data but the stability of the test substance was verified in the diet, over one month for both concentrations.
Duration of treatment / exposure:
3 months
Frequency of treatment:
7 days/week
Remarks:
Doses / Concentrations:
6000, 18000 and 54 000 ppm (approx. 150, 450 and 1350 mg/kg bw/day) for both sexes
Basis:
other: Diet mix test substance concentrations confirmed by analysis
No. of animals per sex per dose:
3
Control animals:
yes, plain diet
Details on study design:
-Dose selection rationale: no
-Rationale for animal assignment (if not random): no data
-Rationale for selecting satellite groups: no
-Post-exposure recovery period in satellite groups: no
-Section schedule rationale (if not random): /
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked in table [No.7.5.1-2] were included.


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily


BODY WEIGHT: Yes
- Time schedule for examinations: weekly


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes


FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data


OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: once before treatment, then after 1 and 3 months. Examination by indirect ophthalmoscopy and/or retinoscopy
- Dose groups that were examined: all groups


HAEMATOLOGY: Yes
- Time schedule for collection of blood: 1 week before treatment, 4,8 and 12-weeks after treatment
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 3 males and 3 females
- Parameters checked in table [No. 7.5.1-3] were examined.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: 1 week before treatment, 4,8 and 12-weeks after treatment
- Animals fasted: No data
- How many animals: 3 dogs/sex
- Parameters checked in table [No. 7.5.1-3] were examined.


URINALYSIS: Yes
- Time schedule for collection of urine: 1 week before treatment, 4,8 and 12-weeks after treatment
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.7.5.1-3] were examined.


NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations: /
- Dose groups that were examined: /
- Battery of functions tested: sensory activity / grip strength / motor activity / other: /


OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table 7.5.1-4)
HISTOPATHOLOGY: Yes (see table 7.5.1-4)

- included organ examination for tongue, oesophagus, stomach, small intestine, colon, liver, pancreas, salivary glands, trachea, lungs, heart, aorta, spleen, mesenteric and popliteal lymph nodes, thymus, thyroid, parathyroids, adrenals, pituitary, striated muscle, kidneys, bladder, gonads, epididymis, prostate, uterine horns, brain, spinal cord, eyes and optic nerves.
- included organ weight for liver, lungs, thyroid, adrenals, heart, spleen, kidneys, prostate, brain, gonads and uterine horns.
Statistics:
Analysis of variance followed by Student's "t" test or analysis of covariance for the mean absolute organ weight.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY: no deaths were observed and all animals showed a normal appearance and behaviour through out the treatment period.


BODY WEIGHT AND WEIGHT GAIN: no effects by comparison with negative control animals


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): no abnormality was observed


FOOD EFFICIENCY: no data


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): not applicable


OPHTHALMOSCOPIC EXAMINATION: no abnormality was observed


HAEMATOLOGY: no significant abnormality was observed


CLINICAL CHEMISTRY: no significant abnormality was observed


URINALYSIS: no abnormality was observed


NEUROBEHAVIOUR: not examined


ORGAN WEIGHTS: a slight but significant (P=0.05) increase in absolute liver weights in the 2 highest dose treated dogs as compared with controls. This was not associated with any abnormality on histological examination.


GROSS PATHOLOGY: no effects related with the test substance administration at both doses


HISTOPATHOLOGY: NON-NEOPLASTIC: no abnormality was observed. In particular, the hepatic parenchyma of those animals showing increase liver weights, was histologically normal.


HISTOPATHOLOGY: NEOPLASTIC (if applicable): /


HISTORICAL CONTROL DATA (if applicable)


OTHER FINDINGS
Dose descriptor:
NOAEL
Effect level:
>= 1 350 mg/kg bw/day (actual dose received)
Sex:
male/female
Critical effects observed:
not specified

Table 7.5.1 -5:   Mean food consumption (in g/day)

Conditions

Negative control

Low dose

6000 ppm

Mid dose

18 000 ppm

High dose

54 000 ppm

Before treatment

Week 3 (or 2*)

250

250

250

250

Week 1

250

250

250

250

During treatment

Week 1

250

250

250

250

Week 2

250

250

250

245

Week 3

250

250

250

250

Week 4

250

250

250

248

Week 5

250

250

250

243

Week 6

250

250

250

241

Week 7

250

250

250

245

Week 8

250

250

250

248

Week 9

250

250

250

250

Week 10

250

250

250

250

Week 11

250

250

250

250

Week 12

250

250

250

250

*Significantly different (p 0.05) from the control

**Significantly different (p 0.01) from the control

Table 7.5.1 -6:   Mean bodyweights (in kg)

Conditions

Negative control

Low dose

6000 ppm

Mid dose

18 000 ppm

High dose

54 000 ppm

Before treatment

Week 3

10

8.9

9.3

9.7

Week 1

10.1

8.8

9.3

9.7

During treatment

Week 1

10.2

8.8

9.5

9.8

Week 2

9.2

8.7

9.2

9.5

Week 3

9.9

8.7

9.3

9.4

Week 4

10.0

8.9

9.3

9.5

Week 5

10.1

8.9

9.4

9.4

Week 6

10.1

9.0

9.4

9.6

Week 7

10.4

9.1

9.5

9.8

Week 8

10.4

9.3

10.0

9.9

Week 9

10.8

9.4

10.2

10.1

Week 10

10.6

9.5

10.3

10.3

Week 11

10.9

9.2

10.3

10.3

Week 12

10.7

9.2

10.2

10.2

*Significantly different (p 0.05) from the control

**Significantly different (p 0.01) from the control

Table 7.5.1 -7:   Mean rectal temperature (in °C)

Conditions

Negative control

Low dose

6000 ppm

Mid dose

18 000 ppm

High dose

54 000 ppm

Before treatment

Week 3

38.8

38.8

38.7

38.9

Week 1

38.4

38.8

38.7

38.7

During treatment

Week 1

38.4

38.5

38.3

38.5

Week 2

38.4

38.4

38.3

38.5

Week 3

38.3

38.5

38.4

38.5

Week 4

38.8

38.8

38.8

38.7

Week 5

38.7

38.7

38.6

38.7

Week 6

38.7

38.6

38.6

38.7

Week 7

38.7

38.6

38.7

38.7

Week 8

38.7

38.7

38.9

38.8

Week 9

38.6

38.8

38.9

38.7

Week 10

38.6

38.7

38.7

38.7

Week 11

38.4

38.6

38.4

38.7

Week 12

38.7

38.7

38.7

38.7

Haematological analysis was performed up to 12 weeks after treatment. In the absence of abnormalities, only results observed up to 4 weeks after treatment were reported in the following table (but all data were presented in the report).

Table 7.5.1 -8:   Haematological results before (1 week) and after (4weeks) treatment

Conditions

Circulating blood

Platelets

Clotting time

Hb*

(g/100 mL)

mHc*

(%)

Erythrocytes

(106/mm3)

Leucocytes

(103/mm3)

Differential count

R*

Neutrophils

(%)

Eosinophils

(%)

Basophils

(%)

Monocytes

(%)

Lymphocytes

(%)

NRC*

Howell time

(s)

Quick time

(s)

Thrombin time

(s)

1 week before treatment

Negative control group

16.2

51

6.59

18.8

61

5

0

4

30

0

1.1

540

1.31

7

17

Low dose group

6000 ppm

16.4

49

6.54

19.3

64

6

0

3

27

0

1.1

556

1.26

7

19

Mid dose group

18 000 ppm

16.1

49

6.45

19.9

65

4

0

3

28

0

1.3

563

1.34

7

19

High dose group

54 000 ppm

15.9

47

6.58

18.9

61

5

0

3

31

0

0.9

545

1.54

7

19

4 weeks after treatment

Negative control group

15.3

45

6.46

14.8

60

8

0

2

30

0

0.9

596

1.32

7

19

Low dose group

6000 ppm

15.3

46

6.85

16.5

64

5

0

2

29

0

0.6

548

1.38

7

18

Mid dose group

18 000 ppm

14.5

45

6.32

16.4

62

4

0

3

31

0

0.9

538

1.31

7

18

High dose group

54 000 ppm

15.4

45

6.49

17.6

59

3

0

3

35

0

0.5

482

1.43

7

18

* Hb: haemoglobin; mHc: microhaematocrit; NRC: Nucleated red cells / 100 leucocytes; R: reticulocytes / 100 erythrocytes

Conclusions:
NOAEL = 1350 mg/kg b.w./day
Executive summary:

In a subchronic toxicity study conducted before guidelines, Stearoybenzoylmethane (35 558 RP) was administered daily through diet for 90 days to Beagle dogs (3 dogs/sex/dose) at dose levels of 150, 450 and 1350 mg/kg bw/day (corresponding to 6000, 18 000 and 54 000 ppm respectively). Control group (3 dogs/sex) received diet without test substance.
 Examinations for mortality, clinical signs,  food consumption and body weight gain were performed during the 90-day study period once weekly for all groups. All surviving animals were necropsied at the end of observation period. Hematology, blood chemistry, urinalysis and organ weights (absolute and relative) were determined. Gross and histopathologic pathology were conducted.

All animals appeared normal following all dosing during the period of the study. Body weight and food consumption were unaffected by the test substance administration.
Clinical chemistry analysis and gross pathological analysis did not show any abnormalities in all treated dogs by comparison with controls. At the two highest doses, a slight but significant (P=0.05) increase in absolute liver weights in treated dogs as compared with controls. However, this increase was not associated with any abnormality on histological examination.

The NOAEL is equal to or higher than 1350 mg/kg b.w./day.

This subchronic toxicity study in the dog is considered acceptable as an alternative to a subchronic toxicity study in the rat (OECD 408), even if it doesn't satisfy all the guideline requirements for a subchronic oral study in dogs in current OECD 409 guidelines.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 350 mg/kg bw/day
Study duration:
subchronic
Species:
dog
Quality of whole database:
The study was conducted according to scientific principles of guideline OECD 409 (Kimish score = 2)

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The repeated-dose toxicity of the reaction mass of Stearoylbenzoylmethane and Palmitoylbenzoylmethane was tested by the oral route in rats (for up to 2 years) and in dogs (for 90 days).

Two study of reliability 2 according to Klimisch criteria are available and were selected as key studies (90-day dog study and 28-day rat study; Ganter et al. 1977). In the dog study, the test substance was administered daily through diet for 90 days to Beagle dogs at dose levels of 150, 450 and 1350 mg/kg bw/day. All animals appeared normal following all dosing during the study period. Body weight and food consumption were unaffected by the test substance administration. Clinical chemistry analysis and gross pathological analysis did not show any abnormalities in all treated dogs by comparison with controls. At the two highest doses, a slight but statistically significant increase in absolute liver weights was noted in treated dogs as compared with controls. However, this increase was not associated with any abnormality on histological examination. Therefore the NOAEL was considered to be equal to or higher than 1350 mg/kg b.w./day.

In the 28-day rat study, the test substance was administered daily through diet for 28 days to CD rats at dose levels of 2000 and 10000 mg/kg bw/day. All animals appeared normal following both dosing throughout the study period. Body weight and food consumption were unaffected by the test substance administration. Clinical chemistry analysis and gross pathological analysis including absolute and relative organ weight did not show any abnormalities in all treated rats by comparison with controls. Therefore the NOAEL was considered to be equal or higher than 10 000 mg/kg b.w./day.

The absence of relevant toxicity in rats was confirmed in the third study (Mossinger, 1977) assigned of reliability 3 owing to significant deviations to the technical guidelines currently in force (e.g. only one dose tested instead of at least three) and used as part of a weight of evidence approach. In this study, the test substance was administered in the diet for 24 months to Wistar rats (20/sex/dose) at a single dose level of 100 ppm (corresponding to approximately 5 mg/kg bw/day). Two treated males died during the exposure period: one for unknown reasons while incipient acute mycoplasmic bronchopneumonia was observed in the second animal. One female also died during the exposure and chronic typhlitis was observed at necropsy. No haematological abnormalities were observed. No differences were reported in incidence of any non-neoplastic lesions at macroscopic and microscopic pathological examinations when compared to controls.

No repeated dose toxicity data specific to the reaction mass of Stearoylbenzoylmethane and Palmitoylbenzoylmethane are available for the dermal and inhalative routes of exposure.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The study with the longest duration (90 days) and the lowest NOAEL was chosen.

Justification for classification or non-classification

There is no evidence for intrinsic toxic properties of the reaction mass of Stearoylbenzoylmethane and Palmitoylbenzoylmethane relevant to humans obtained from the results of reliable, relevant and adequate subacute and subchronic oral toxicity studies in rats and dogs, respectively. Therefore, no classification is required for repeated dose toxicity according to Directive 67/548/EEC or CLP, Regulation (EC) No 1272/2008 with respect to the systemic toxicity or Specific Target Organ Toxicity, repeated administrations.