Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 August 2014 to 24 March 2016
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP status of study not documented.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Deviations:
no
GLP compliance:
not specified
Remarks:
The study was conducted in compliance with Metrology Accreditation of P.R.C.
Limit test:
no

Test material

Constituent 1
Test material form:
solid
Details on test material:
- Appearance: black solid
- Storage conditions: at room temperature, kept in a dry place and sealed

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Zhejiang Center of Laboratory Animals, Hangzhou, Zhejiang
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 4 - 5 weeks old
- Weight at study initiation: 186 – 244 g (the intragroup weight variation did not exceed ±10 % of the mean weight between-groups; the intergroup weight variation did not exceed ± 5 % of the mean weight between-groups)
- Housing: animals were housed in pairs, by sex in suspended, wire bottom, stainless steel cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 3 °C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 12 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours darek / 12 hours light

Administration / exposure

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Remarks:
the test material was mixed with distilled water (mass ratio of 1:1.1) to form a paste
Details on exposure:
TEST SITE
- Area of exposure: the dorsal surface of the trunk of each animal was clipped free of fur on the day prior to treatment
- % coverage: 10% (minimum)
- Type of wrap if used: the area of application was covered with a cellophane patch and secured with non-irritating adhesive tape. The rats were fixed with rat fixator to prevent possible ingestion of the test material.
- Time intervals for shavings or clipplings: at least once a week

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test material was gently washed with tap water
- Time after start of exposure: 6 hours after application

TEST MATERIAL
- For solids, paste formed: yes
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
316 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
2 groups of 15 males and 15 females received applications of 0 (control) and 1000 mg/kg bw/day

2 groups of 10 males and 10 females received applications of 100 and 316 mg/kg bw/day test material
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale:
Doses were based on the acute dermal LD50 values (> 20000 mg/kg bw/day in male and female rats)

- Post-exposure recovery period in satellite groups: Ten rats (five per sex per group) in the control and high groups (two additional satellite groups) were observed for at least 14 days post treatment, for observation of reversibility or persistence of any toxic effects

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: animals were observed twice daily for signs of morbidity/mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once before the first exposure and at least once a week thereafter
Signs noted included, but not be limited to, changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling) or bizarre behaviour (e.g. self-mutilation, walking backwards) were recorded.

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Once before the first exposure and at least once a week thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: prior to the initiation of the study, weekly thereafter, and at death/or sacrifice

FOOD CONSUMPTION: Yes

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule: Ophthalmological examination was made prior to the administration of the test material and at the termination of the study

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the study, blood samples were obtained from abdominal aorta of each rat
- Anaesthetic used for blood collection: Yes (10% chloral hydrate )
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked included: white blood cell (WBC), red blood cell (RBC), haemoglobin (HGB), haematocrit (HCT) , lymphocyte (LYM), granulocyte (GRA) , monocyte (MID), Eosinophil (EOS), basophil (BAS), red blood cell volume distribution width (RDW), platelet (PLT), prothrombin time (PT), activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the study, blood samples were obtained from abdominal aorta of each rat
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked included: total bilirubin (BIL-T), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP), albumin (ALB), globulin (GLB), alkaline phosphatase (ALP), urea nitrogen (BUN), creatinine (CREA), glucose(GLU), potassium (K), sodium (Na), chlorine (Cl), calcium (Ca)

URINALYSIS: Yes
At the end of the study, urinalyses were conducted from each rat and results were read using CF-U180 urine analyser. Parameters measured included appearance (colour and clarity), glucose (uGLU), bilirubin (uBIL), ketone (KET), occult blood (RBC), acidity (PH), protein (PRO), urobilinogen (uBG), nitrite (NIT), and leukocytes (LEU).
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals in the study was subjected to a full, detailed gross necropsy which includes careful examination of the external surface of the body, all orifices, and the cranial, thoracic and abdominal cavities and their contents.

ORGAN WEIGHTS: Yes
The liver, kidneys, adrenals, testes, epididymis, uterus, ovaries, thymus, spleen, brain and heart of all animals were trimmed of any adherent tissue, as appropriate, and their wet weight taken as soon as possible. Organ coefficients were calculated by expressing organ weight as a percentage of body weights.

HISTOPATHOLOGY: Yes
Tissues and organs (all gross lesions, brain, spinal cord, stomach, thyroid, thymus, small and large intestines, pancreas, liver, kidneys, adrenals, spleen, heart, trachea and lungs, gonads, uterus, accessory sex organs, prostate, urinary bladder, lymph nodes, peripheral nerve, bone marrow, skin) samples from control and treated rats were cut into thin slices, and then fixed into 4% formaldehyde for more than one week. The tissues and organs were then processed through a graded series of ethanol and xylene, and embedded in paraffin. Organs and tissues sections were stained with hematoxylin and eosin (H&E) for histopathological examination under a microscope.
Statistics:
A parametric or non-parametric test was selected based on the results of normality test and homogeneity of variance test. One-way analysis of variance and Dunnett’s t test were used in parameter test. Kruskal-Wallis rank sum test and Wilcoxon-Wilcox rank sum test were used in non-parametric test. Student’s t test was adopted by comparing data between additional high dose and additional control group, or nonparametric Wilcoxon test. Fisher's exact probability test was used for enumeration data. The BMDS2.50 (US EPA) software was used to calculate benchmark dose.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
no systemic clinic signs were observed throughout the study period.
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
During exposure period, erythema and scab were observed on the skin of rats in animals of the high dose group (15 males and 15 females), intermediate dose group (8 males and 8 females), and in the low dose group (2 males and 5 females). Compared with the control group, there were statistically significant differences in all groups of rats except for the low dose group of male rats. Skin irritation symptoms in the high dose group disappeared 7 days later after exposure ended.
No oedema was noted.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
During exposure period, for male rats, body weight gains in the low and high dose group was lower than the control group, the differences both had statistical significance (p < 0.05), but with no obvious dose-effect relationship (Spearman rank correlation: correlation coefficient rs = 0.2283, p = 0.1920).
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effects on food consumption were noted during the exposure period or during the recovery period.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
Ophthalmological examination was made prior to the administration of the test material and at the termination of the study. The results showed that no abnormal changes were found in each dose group for males and females.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
At the end of exposure period, for female rats, WBC, GRA, EOS, BAS of the high dose group animals were higher than for the animals in the control group, LYM, PT of high dose group were lower than the control group; for male rats, RDW, APTT of high dose group was higher than the control group, WBC was lower than the control group (p < 0.05).
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
For female rats, GLU of low dose group was increased, Cl-, Na+ of the intermediate dose group were increased and ALB, A/G were decreased; Cl-, Na+, Ca+ of high dose group were increased and ALB, A/G were decreased at the end of the exposure; ALB,Cl-, Na+, Ca+ of high dose group were increased and A/G was decreased at the end of additional observation. For male rats, AST, ALT of low dose group were increased and Ca+ was decreased, ALT, Cl-, Na+, BUN of high dose group were increased at the end of the exposure; A/G of high dose group were decreased and ALB was increased at the end of additional observation, the differences had statistical significance, but no obvious dose-effect relationship and/or biological significance were found.
Urinalysis findings:
no effects observed
Description (incidence and severity):
Compared the treatment groups with the control group, the differences had no significance (p > 0.05).
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
At the end of exposure period, for female rats, the organ weight and the organ coefficient (liver, adrenal) of high dose group were higher than control group, the differences both had statistical significance (p < 0.05). In addition, the organ coefficient of the brain in the low dose group was higher than in the control group, the differences had statistical significance (p < 0.05), but no obvious dose-effect relationship was found.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No abnormalities were observed at gross necropsy.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No abnormal changes were observed in any treated groups compared with the control group.
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined

Effect levels

open allclose all
Dose descriptor:
BMDL10
Effect level:
11.35 mg/kg bw/day
Based on:
test mat.
Sex:
male
Basis for effect level:
dermal irritation
Dose descriptor:
BMDL10
Effect level:
15.41 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
dermal irritation
Dose descriptor:
NOAEL
Remarks:
systemic effects
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical biochemistry
Dose descriptor:
NOAEL
Remarks:
systemic effects
Effect level:
316 mg/kg bw/day
Based on:
test mat.
Sex:
male
Basis for effect level:
clinical biochemistry

Target system / organ toxicity

Critical effects observed:
no

Any other information on results incl. tables

Table 1: The statistics of local skin irritation signs

Sex

Group

Number

erythema, scab

Number

 

average credit

Female

Control

15

0

 

0.00 ± 0.00

 

Low dose

10

5#

 

1.10 ± 1.16*

 

Intermediate dose 

10

8#

 

2.17 ± 1.15*

 

High dose 

15

15#

 

3.32 ± 0.15*

Male

Control

15

0

 

0.00 ± 0.00

 

Low dose

10

2

 

0.19 ± 0.52

 

Intermediate dose 

10

8#

 

1.69 ± 1.17*

 

High dose 

15

15#

 

3.09 ± 0.35*

Note:

# Fisher probabilities, compared with the control group, P < 0.0170

*Dunnett’s t testcompared with the control group, P < 0.05

Table 2: Skin irritation (erythema) gradings

Number

Sex

Group

Observation timeweek

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

F1

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F2

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F3

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F4

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F5

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F6

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F7

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F8

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F9

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F12

Female

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F13

Female

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

F14

Female

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

F15

Female

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

F16

Female

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

F17

Female

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

F18

Female

Low

0

0

0

0

1

1

1

2

3

4

4

4

4

4

-

-

-

-

F19

Female

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F23

Female

Low

0

0

0

1

1

1

2

2

4

4

4

4

4

4

-

-

-

-

F24

Female

Low

0

0

0

0

1

1

2

2

4

4

4

4

4

4

-

-

-

-

F25

Female

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F26

Female

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F27

Female

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F28

Female

Low

0

0

0

1

1

1

2

4

4

4

4

4

4

4

-

-

-

-

F29

Female

Low

0

0

0

0

1

1

2

4

4

4

4

4

4

4

-

-

-

-

F34

Female

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

Number

Sex

Group

Observation timeweek

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

F35

Female

Intermediate

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F36

Female

Intermediate

0

0

1

0

1

2

4

4

4

4

4

4

4

4

-

-

-

-

F37

Female

Intermediate

0

0

1

1

1

2

4

4

4

4

4

4

4

4

-

-

-

-

F38

Female

Intermediate

0

0

1

1

2

4

4

4

4

4

4

4

4

4

-

-

-

-

F39

Female

Intermediate

0

0

0

1

1

2

4

4

4

4

4

4

4

4

-

-

-

-

F45

Female

Intermediate

0

0

1

1

0

3

4

4

4

4

4

4

4

4

-

-

-

-

F46

Female

Intermediate

0

0

0

2

2

3

4

4

4

4

4

4

4

4

-

-

-

-

F47

Female

Intermediate

0

0

1

1

2

4

4

4

4

4

4

4

4

4

-

-

-

-

F48

Female

Intermediate

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

F49

Female

Intermediate

0

0

1

0

2

4

4

4

4

4

4

4

4

4

-

-

-

-

F56

Female

high

0

2

1

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F57

Female

high

0

2

1

3

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F58

Female

high

0

2

1

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F59

Female

high

0

0

2

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F67

Female

high

0

2

4

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F68

Female

high

0

0

1

3

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F69

Female

high

0

1

2

3

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F78

Female

high

0

1

2

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F79

Female

high

0

0

2

3

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F89

Female

high

0

1

1

2

4

4

4

4

4

4

4

4

4

4

-

-

-

-

F123

Female

Additional high

0

1

1

2

4

4

4

4

4

4

4

4

4

4

0

0

0

0

F124

Female

Additional high

0

2

4

4

4

4

4

4

4

4

4

4

4

4

0

0

0

0

F125

Female

Additional high

0

2

4

4

4

4

4

4

4

4

4

4

4

4

0

0

0

0

F126

Female

Additional high

0

1

2

4

4

4

4

4

4

4

4

4

4

4

0

0

0

0

F127

Female

Additional high

0

0

1

3

4

4

4

4

4

4

4

4

4

4

0

0

0

0

Number

Sex

Group

Observation timeweek

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

M1

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M2

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M3

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M4

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M5

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M6

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M7

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M8

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M9

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M12

Male 

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M13

Male 

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

M14

Male 

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

M15

Male 

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

M16

Male 

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

M17

Male 

Additional control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

M18

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M19

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M23

Male 

Low

0

0

0

0

0

0

1

1

1

0

0

0

0

0

-

-

-

-

M24

Male 

Low

0

0

0

0

0

0

1

1

3

3

3

4

4

4

-

-

-

-

M25

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M26

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M27

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M28

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M29

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M34

Male 

Low

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

 

Number

Sex

Group

Observation timeweek

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

M35

Male 

Intermediate

0

0

0

0

0

2

3

4

4

4

4

4

4

4

-

-

-

-

M36

Male 

Intermediate

0

0

0

0

0

2

3

4

4

4

4

4

4

4

-

-

-

-

M37

Male 

Intermediate

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M38

Male 

Intermediate

0

0

0

0

1

1

3

4

4

4

4

4

4

4

-

-

-

-

M39

Male 

Intermediate

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M45

Male 

Intermediate

0

0

0

0

1

2

4

4

4

4

4

4

4

4

-

-

-

-

M46

Male 

Intermediate

0

0

0

0

1

2

4

4

4

4

4

4

4

4

-

-

-

-

M47

Male 

Intermediate

0

0

0

0

2

2

3

4

4

4

4

4

4

4

-

-

-

-

M48

Male 

Intermediate

0

0

0

0

0

1

3

4

4

4

4

4

4

4

-

-

-

-

M49

Male 

Intermediate

0

0

0

0

0

0

0

0

0

0

0

0

0

0

-

-

-

-

M56

Male 

high

0

0

0

1

1

4

4

4

4

4

4

4

4

4

-

-

-

-

M57

Male 

high

0

0

0

1

1

4

4

4

4

4

4

4

4

4

-

-

-

-

M58

Male 

high

0

1

3

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

M59

Male 

high

0

0

1

2

2

4

4

4

4

4

4

4

4

4

-

-

-

-

M67

Male 

high

0

1

2

3

4

4

4

4

4

4

4

4

4

4

-

-

-

-

M68

Male 

high

0

1

2

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

M69

Male 

high

0

1

3

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

M78

Male 

high

0

2

4

4

4

4

4

4

4

4

4

4

4

4

-

-

-

-

M79

Male 

high

0

0

1

3

3

3

4

4

4

4

4

4

4

4

-

-

-

-

M89

Male 

high

0

0

1

2

2

4

4

4

4

4

4

4

4

4

-

-

-

-

M123

Male 

Additional high

0

1

2

4

4

4

4

4

4

4

4

4

4

4

0

0

0

0

M124

Male 

Additional high

0

0

0

1

1

4

4

4

4

4

4

4

4

4

0

0

0

0

M125

Male 

Additional high

0

2

4

4

4

4

4

4

4

4

4

4

4

4

0

0

0

0

M126

Male 

Additional high

0

0

0

2

1

2

4

4

4

4

4

4

4

4

0

0

0

0

M127

Male 

Additional high

0

0

0

3

1

2

4

4

4

4

4

4

4

4

0

0

0

0

 

 

Applicant's summary and conclusion

Conclusions:
The BMDS2.50 (US EPA) software was used to calculate the benchmark dose which was considered to be a suitable replacement to a NOAEL value. Under the conditions of the study the BMDL of the test material in female and male rats were 11.35 mg/kg bw/day and 15.41 mg/kg bw/day, respectively.
Executive summary:

The repeated dose toxicity of the test material was investigated in a study which was conducted under GLP conditions and in accordance with the standardised guideline OECD 411.

During the study 50 Sprague Dawley rats per sex were randomly assigned to four groups: one control group, one low dose group, one intermediate dose group and one high dose group representating test material concentrations of 0.0, 100.0, 316.0, 1000.0mg/kg, respectively. All test animals were exposed to test material and observed for 90 days. Ten rats (five per sex per group) in the control and high groups (two additional satellite groups) were observed for at least 14 days post treatment, for observation of reversibility or persistence of any toxic effects. Clinical signs, mortality, food consumption and body weight change were evaluated. When the study was terminated, urinalyses, gross necropsy, haematology, clinical biochemistry and histopathology were carried out.

Compared with the control group, for female rats the intermediate dose group, Cl- and Na+ were increased while ALB, A/G were decreased. In the high dose group, for female rats, WBC, GRA, EOS, BAS, Cl-, Na+, Ca+, organ weight and organ coefficient (liver, adrenal) were increased, LYM, PT, ALB, A/G were decreased at the end of the exposure period, Cl-, Na+, Ca+ were increased, total food efficiency, A/G were decreased at the end of additional observation period; for male rats, RDW,APTT, Cl-, Na+, BUN were increased, WBC was decreased at the end of exposure period, A/G was decreased at the end of additional observation day. During exposure period,erythema and scab were observed on the skin of rats in the high dose group(15 males and 15 females, the average score of male rats was 3.09 and female rats was 3.32), in intermediate dose group (8 males and 8 females, the average score of male rats was 1.69 and female rats was 2.17), and in the low dose group (2 males and 5 females, the average score of male rats was 0.19 and female rats was 1.10). Compared with the control group, there were statistically significant differences in all groups of rats except low dose group of male rats.Skin irritation symptoms in the high dose group disappeared 7 days later after exposure finished. During exposure and additional 28-day observation period, no systemic clinic signs were observed in rats. There were no abnormal changesin each dose group for males and females on ophthalmological examination prior to the administration of the test material and at the termination of the study. Compared with the control, there was no statistical significance in any treated groups on gross anatomy. At the end of exposure or additional 14-days, no significant histopathological changes were observed in any treated groups compared with control group.

The systemic NOEL was 100 mg/kg bw/day. For local skin irritation effects, the BMDS2.50 (US EPA) software was used to calculate the benchmark dose which was considered to be a suitable replacement to a NOAEL value. Under the conditions of the study the BMDL of the test material in female and male rats were 11.35 mg/kg bw/day and 15.41 mg/kg bw/day, respectively.

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