Phosphoric acid, iron(2+) lithium salt (1:1:1)

Repeated dose toxicity: dermal

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

Test material information

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 / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day

Doses / concentrations 3

Dose / conc.:

316 mg/kg bw/day

Doses / concentrations 4

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

1

Dose descriptor:

BMDL10

Effect level:

11.35 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

dermal irritation

2

Dose descriptor:

BMDL10

Effect level:

15.41 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

dermal irritation

3

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

4

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 test，compared with the control group, P < 0.05

Table 2: Skin irritation (erythema) gradings

Number	Sex	Group	Observation time（week）
			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 time（week）
			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 time（week）
			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 time（week）
			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.