SUMILIZER GP

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 May 1998 - 28 Jan 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Chemical Substances Control Law, Partial amendments of the "Testing Methods of New Chemical Substances"

Version / remarks:

adopted in 1986

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: "Rule to add a New Chemical Substance to the Positive Lists", attachment No. 3, published by JOSPA (Japan Hygenic Olefin and Styrene Plastic Association)

Version / remarks:

adopted in 1992

Test guideline 3

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

adopted in 2018

Deviations:

yes

Remarks:

animals too young (4 weeks), temperature out of range ( 24 ± 2 instead of 22 ± 2 °C), no detailed clinical observation, no ophthalmoscopic or neurobehavioural examination, no T3/T4/TSH levels, no HDL or LDL levels, no histopathology of coagulating gland

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

442-450-4

EC Name:

-

Cas Number:

203255-81-6

Molecular formula:

C42H61O4P

IUPAC Name:

6-(3-(3-tert-Butyl-4-hydroxy-5-methylphenyl)propoxy)-2,4,8,1 0-tetra-tert-butyldibenz(d,f)(1,3,2)dioxaphosphepin

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan, Inc.
- Age at study initiation: 4 weeks
- Weight at dosing: 194.7 - 222.9 g (males) and 140.9 - 175.6 g (females)
- Housing: Individually in stainless steel hanger cages (260 x 200 x 380 mm)
- Diet: MF pellet diet (Oriental Yeast Co., Ltd.) ad libitum
- Water: well water from an automatic watering system, supplied with 2 ppm sodium hypochlorite, ad libitum
- Acclimation period: 13 days (males) and 14 days (females)

DETAILS OF FOOD AND WATER QUALITY:
The diet was analysed by Japan Food Research Laboratories (Tokyo, Japan) and found to contain impurities below the permissible limit. The water was analysed at Kakujo Nakyukaken Center Ltd. (Kumamoto, Japan) and it was verified that impurities were below the permissible limit.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 2
- Humidity (%): 55 ± 10
- Air changes (per hr): 13 - 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 12 May 1998 To: 22 Sep 1998

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5% formulation

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance was suspended in 0.5% methylcellulose solution at the concentrations of 1.0, 3.0 and 10.0% (w/v). The formulations were prepared 2-3 times a week and placed in a brown container and stored at room temperature until use.

VEHICLE
- Concentration in vehicle: 1, 3 and 10% (w/v)
- Amount of vehicle: 10 mL/kg bw/day

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Uniformity tests and stability tests of the test substance in vehicle were conducted at two concentrations (0.1% and 10% w/v, corresponding to 1 and 100 mg/mL), including the concentration range of dosing solutions used in the preliminary dose finding study before start of administration. Samples were taken from the upper, middle and lower layer of the formulation.The tests confirmed that the test substance suspended in 0.5% methylcellulose solution was stable in a brown container stored at room temperature for 8 days and that uniformity was not affected. Concentrations of the dosing solutions at the respective concentrations prepared for the first time were measured to confirm that the concentrations were within the allowable range of ± 10%. Mean concentrations for the 1 and 100 mg/mL formulations were 0.954 and 99.99.789 mg/mL on Day 0, 0.936 and 97.753 mg/mL on Day 5 and 0.903 and 102.619 mg/mL on Day 8. Uniformity of all samples available ranged from 98.1 – 101.8% on Day 0, 98.5 – 100.6% on Day 5 and 98.6 – 101.6% on Day 8.

Duration of treatment / exposure:

42 days (interim sacrifice control and test groups)
90 days (control and test groups)
90 days and 28 days post-exposure observation period (satellite control and high dose group)

Frequency of treatment:

once daily, 7 days/week

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10 (main study)
10 (satellite control and high dose group)
6 (interim sacrifice group)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Dose levels were selected based on the results of a preliminary 2-weeks repeated dose toxicity study, in which male and female rats received an oral dose of 100, 300 and 1000 mg/kg bw/day. No noticable toxicological changes were observed in male or female rats at 1000 mg/kg bw/day. Therefore, 1000 mg/kg bw/day was set as the maximum dose in the study, which was the upper limit under the guideline of Chemical Substances Control Law (MITI, Japan), and 300 and 100 mg/kg bw/day were then set according to the common ratio of about 3.

- Rationale for animal assignment: The rats were allocated to the treatment groups and the control group in the day before start of administration by the stratified sequential randomization method on the basis of their body weights. Further, 6 each of the male and female rats were selected from the rats remaining after the group allocation in such a manner to minimise the standard deviation of body weights with respect to each sex for use for blood collection before start of administration.

- Fasting period before blood sampling for clinical biochemistry: 18 - 24 h

- Post-exposure recovery period in satellite groups: 28 days

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily, before and after administration during the administration period and in the morning and afternoon every day during the recovery period.
- Examinations included: clinical signs and mortality

BODY WEIGHT: Yes
- Time schedule for examinations: Before administration on the day of start of administration, once weekly thereafter, on the day previous to necropsy and on the day of necropsy.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, food consumption was calculated by measuring the amount of diet supplied and that remaining.

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: Yes, efficiency of food conversion was calculated based on food consumption and the weekly increase in body weight.

WATER CONSUMPTION: Yes
- Time schedule for examinations: Water consumption was measured on Day 2 of administration and then once a week and on the day previous to necropsy. Daily water consumption was calculated by measuring the amount of water supplied and that remaining.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Before start of administration, at completion of 42 days of administration, at completion of 90 days of administration and at completion of the recovery period.
- Anaesthetic used for blood collection: Yes (intraperitoneal administration of 30 mg/kg bw sodium pentabarbiturate)
- Animals fasted: Yes, the rats to be employed for blood collection were fasted for about 18 – 24 h.
- How many animals: all animals
- Parameters examined: erythrocyte count, leukocyte count, differential leukocyte count (neutrophil, eosinophil, basophil, lymphocyte and monocyte), hemoglobin, hematocrit, platelet count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MHC), mean corpuscular hemoglobin concentration (MCHC), microscopical examination of blood smear, prothrombin time and activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Before start of administration, at completion of 42 days of administration, at completion of 90 days of administration and at completion of the recovery period.
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: total protein, albumin, A/G ratio, total bilirubin, GOT, GPT, -glutamyl transpeptidase (-GTP), alkaline phosphatase (ALP), total cholesterol, triglyceride, glucose, urea nitrogen (BUN), creatinine, inorganic phosphorus (IP), calcium (Ca), sodium (Na), potassium (K) and chloride (Cl)

URINALYSIS: Yes
- Time schedule for collection of urine: The urine was collected during the period for 8 to 12 o’clock in the morning (the period before administration during the administration period) before administration, in Weeks 6 and 13 of administration and in Week 4 of recovery.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- How many animals: Urinalysis was conducted for all male and female rats before start of administration, for 10 male and female rats each in the groups for which administration was terminated after 90 days in Weeks 6 and 13 of treatment, and for 10 male and female rats each in the respective groups in the recovery study in Week 4 of recovery.
- Parameters examined: Fresh urine was examined for pH, protein, glucose, keton bodies, bilirubin, occult blood and urobilinogen. Urine accumulated for 24 h was collected under conditions of diet and water supply after the collection of fresh urine and examined for color tone.

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes. At the times of completion of 90 days of administration and completion of the recvoery period, the rats were euthanized by exsanguination and necropsied. All organs and tissues were observed macroscopically.
The following organs were weighed: brain, pituitary, thyroids (including the parathyroid glands), heart, lungs (including bronchi), thymus, liver, spleen, kidneys, adrenals, testes and ovaries

HISTOPATHOLOGY: Yes.
The following organs were collected and fixed for histophatological examination: brain, pituitary, thyroids (including the parathyroid glands), heart, lungs (including bronchi), thymus, liver, spleen, kidneys, adrenals, testes, ovaries, spinal cord (thoracic region), eyes, Harderian gland, submandibular lymph nodes, salivary glands (sublingual gland and submandibular gland), larynx, trachea, tongue, pancreas, esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, mesenteric lymph nodes, urinary bladder, seminal vesicles, prostate, epididymides, uterus, vagina, femur (including bone marrow), sternum (including bone marrow), skin (lower abdomen), mammary glands (female only), aorta (thoracic region), sciatic nerve, biceps femoris muscle and tissues with gross lesions
For the brain, pituitary, thyroids, heart, lungs, thymus, liver, pancreas, spleen, kidneys, adrenals, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, submaxillary lymph nodes, mesenteric lymph nodes, bone marrow, testes, ovaries and tissue with gross lesions of the rats in the respective groups collected at completion of 90 days of administration, paraffin sections were prepared, stained with hematoxylin-eosin staining, and examined microscopilcally. For a part of the male rats in the control group and the maximum dose group, PAS staining was performed for the kidneys. Since increases in the weights of the thyroids, liver, and kidneys were observed in the maximum dose group at completion of the recovery period, these organs from male and female rats in control and maximum dose groups were microscopically examined. In addition, the tissues with gross lesions of the respective groups were also examined microscopically.

Statistics:

Statistical analyses were performed for the body weight, food consumption, efficiency of food conversion, water consumption, urinalysis, hematological examination, blood biochemical examination, organ weights, the ratio of organ weight to body weight, necropsy and histopathological examination. Numerical data were compared the variance among groups of data. Bartlett’s test was used to compare the variance among groups of data. If the groups were accepted to be homogeneous, Dunnett’s multiple comparison test was used for comparison of groups of data. If the groups of data were heterogeneous in Bartlett’s test, Steel’s multiple comparison test was used. A cumulative chi-square test was used for comparison of the results of the urinalysis, a Fisher’s accuracy establishment test for comparison of the results of necropsy, and Mann-Whitney’s U-test for comparison of the results of the histopathological examination with those of the control group. In all cases, the levels of p < 0.01 and p < 0.05 were considered to be significant, and two-sided tests were performed.

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Hair loss was noted in 1/26 males of the control group and 1/26 males of the 1000 mg/kg bw/day group and in 1/26 females of the 100 mg/kg bw/day group during the administration period or the recovery period. The observation was considered to be incidental.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

100 mg/kg bw/day: statistically significant increase in absolute food consumption on Day 57 in males (28.5 ± 2.8 vs. 25.7 ± 2.2 g/rat/day in the control group)
300 mg/kg bw/day: statistically significant decrease in absolute food consumption on Day 64 in females (16.0 ± 2.1 vs. 18.2 ± 2.3 g/rat/day in the control group)
1000 mg/kg bw/day: statistically significant increase in absolute food consumption on Day 15 in males (28.2 ± 2.3 vs. 26.5 ± 2.3 g/rat/day in the control group); statistically significant increase in absolute food consumption (28.8 ± 2.8 vs. 25.7 ± 2.2 g/rat/day in the control group) and food consumption per body weight (5.7 ± 0.3 % vs. 5.3 ± 0.4 % in the control group) on Day 57 in males; statistically significant decrease in absolute food consumption (15.0 ± 2.2 vs. 18.2 ± 3.1 g/rat/day in the control group) and in food consumption per body weight (4.9 ± 0.8 % vs. 5.6 ± 0.6 % in the control group) on Day 22 of recovery in females
All effects were transient, without dose-response relationship and/or occurred only during the recovery period and not during treatment and were therefore considered not treatment-related.

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

effects observed, non-treatment-related

Description (incidence and severity):

100 mg/kg bw/day: statistically significant decrease in absolute water consumption (27.2 ± 6.8 vs. 33.8 ± 5.6 g/rat/day in the control group) and water consumption per body weight (9.9 ± 2.4% vs. 11.9 ± 2.0% in the control group) on Day 64 in females
1000 mg/kg bw/day: tendency towards decrease in absolute water consumption during the recovery period in females (please refer to Table 1 under “Any other information on results incl. tables”).
The changes were transient, not dose-dependent and/or occurred only during the recovery period but not during treatment and were therefore considered to be incidental but not related to treatment.

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

100 mg/kg bw/day: statistically significant decrease in hemoglobin (-4.5%, 13.9 ± 0.4 g/dL vs. 14.4 ± 0.5 g/dL in the control group) in females after 90 days of administration
1000 mg/kg bw/day: statistically significant decreases in hemoglobin (-4.2%, 13.8 ± 0.3 g/dL vs. 14.4 ± 0.5 g/dL in the control group) and hematocrit (-3-3%, 44.2 ± 1.1 % vs. 45.7 ± 1.1 % in the control group) and statistically significant increase in platelet count (+16.6%, 105.4 ± 13.3 x 10E4/µL vs. 90.4 ± 5.7 x 10E4/µL in the control group) in females after 90 days of administration.
All changes were slight, within the range of physiological changes specified in historical control data and not accompanied by changes in other erythrocyte or blood coagulation parameters and therefore considered to be incidental but not related to treatment.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

100 mg/kg bw/day:
- statistically significant increase in glucose (+23.5%, 126 ± 20 mg/dL vs. 102 ± 12 mg/dL in the control group) in males of the interim sacrifice group after 42 days of treatment, not treatment-related
- statistically significant increase in inorganic phosphorus (+15.2%, 7.6 ± 0.6 mg/dL vs. 6.6 ± 0.9 mg/dL in the control group) in females of the interim group after 42 days of treatment, not treatment-related
- statistically significant increase in glucose (+17.8%, 139 ± 15 mg/dL vs. 118 ± 7 mg/dL in the control group) in males after 90 days of treatment, not dose-related, within historical background data and without histopathological abnormalities in glucose metabolism regulating organs

300 mg/kg bw/day:
- statistically significant increase in total cholesterol (+44.2%, 75 ± 16 mg/dL vs. 52 ± 10 mg/dL in the control group) in females of the interim sacrifice group, not treatment-related
- statistically significant increase in glucose (+14.4%, 135 ± 8 mg/dL vs. 118 ± 7 mg/dL in the control group) in males after 90 days of treatment, not dose-related, within historical background data and without histopathological abnormalities in glucose metabolism regulating organs
- statistically significant decrease in GPT (-19%, 17 ± 2 IU/L vs. 21 ± 4 IU/L in the control group) in females, not dose-related and within historical background data

1000 mg/kg bw/day:
- statistically significant decrease in sodium (-0.7%, 145.6 ± 0.9 mEq/L vs. 146.6 ± 0.6 mEq/L in the control group) in females of the interim sacrifice group after 42 days of treatment, not treatment-related as no change was observed after 90 days of treatment in the main group
- statistically significant increase in total cholesterol (+38.2%, 94 ± 20 mg/dL vs. 68 ± 14 mg/dL in the control group) in females after 90 days of treatment, toxicologically not relevant due to the absence of histopathological changes and full reversibility after the 28-day recovery period
- statistically significant increase in glucose (+13.5%, 134 ± 12 mg/dL vs. 118 ± 7 mg/dL in the control group) in males after 90 days of treatment, not dose-related, within historical background data and without histopathological abnormalities in glucose metabolism regulating organs
- statistically significant decreases in alkaline phosphatase (-16.4%, 112 ± 22 IU/L vs. 134 ± 21 IU/L in the control group), blood urea nitrogen (-8.3%, 14.4 ± 1.0 mg/dL vs. 15.7 ± 1.3 mg/dL in the control group) and creatinine (-20%, 0.4 ± 0.1 mg/dL vs. 0.5 ± 0.1 mg/dL in the control group) in males after 28-days of recovery, without toxicological significance as the observations were not made during the treatment-period
- statistically significant decrease in GOT (-20.4%, 74 ± 11 IU/L vs. 93 ± 22 IU/L in the control group) and statistically significant increase in total bilirubin (0.1 ± 0.0 mg/dL vs. 0.0 ± 0.0 mg/dL in the control group) in females after 28-days of recovery, without toxicological significance as the observations were not made during the treatment-period

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

After 90 days of treatment, statistically significant increases were observed in the relative liver weight for females at ≥ 300 mg/kg bw/day (+7.9% and +11.1% at 300 and 1000 mg/kg bw/day, respectively). The effect in females was fully reversible within the 28-days recovery period. In males, absolute and relative liver weights (+10.9% and +11.4%) and absolute kidney weights (+7.2%) after 90 days of treatment were statistically significantly increased at 1000 mg/kg bw/day. The observations were still present at the end of the recovery period. (For details please refer to Table 2 under “Any other information on results incl. tables”.) The observations were considered as treatment-related, but in the absence of histopathological findings or renal changes, the effect was considered to be of less toxicological relevance.
After the recovery period, there were statistically significant increases in absolute (+23.4%) and relative (+19.1%) thyroid weights, statistically significant increases in absolute (+16.9%) and relative (+14.2%) spleen weight and statistically significant increases in absolute lung weight (+8.2%) in males at 1000 mg/kg bw/day. In females of the high dose recovery group, statistically significant decreases in the absolute (-13.8%) and relative (-11.1%) heart weight were noted. (For details please refer to Table 3 under “Any other information on results incl. tables”.) The findings in the recovery group animals were slight, well within historical background ranges and not observed during the treatment period and therefore considered non-adverse.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

100 mg/kg bw/day: light gray nodule in the femur of 1/10 males and loss of hair in 1/10 females after 90 days of treatment, not treatment-related.
1000 mg/kg bw/day: light yellow nodule in the epididymides in 1/10 males after 90 days of treatment, not treatment-related; dark red macula in the lung of 1/10 females and loss of hair in 1/10 males at the end of the recovery period, not treatment-related.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

100 mg/kg bw/day: After 90 days of treatment, hyperplasia of the squamous cells in the skin in was noted in 1/10 females (evident as hair loss at macroscopical examination), localized hemorrhage in the liver was observed in 1/10 males and osteochondroma was noted in 1/10 males (appearing as light gray nodule in the femur at macroscopical examination). As no dose-response relationship was evident, the findings were considered not treatment-related.
1000 mg/kg bw/day: After 90 days of treatment, spermatic granuloma in the epididymides (appearing as light yellow nodule in macroscopic examination) was observed in 1/10 male rats. The finding is a commonly observed spontaneous lesion and therefore considered not treatmend-related. In the recovery animals, localized hemorrhage was noted in the alveoli in 1/10 females (appearing as dark red macula in the lung at macroscopical examination). The finding was considered not treatment-related because it was observed in 1 rat only and was not observed during the treatment.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Target system / organ toxicity

Key result

Critical effects observed:

no

Any other information on results incl. tables

Table 1: Water consumption in female rats treated for 90 days followed by 28 -day withdrawal (recovery) period

	Days of withdrawal
	1	8	15	22	28
Control
n	10	10	10	10	10
mean ± SD	33.9 ± 6.2	34.8 ± 6.4	36.9 ± 2.3	36.9 ± 7.1	34.8 ± 6.5
1000 mg/kg bw/day
n	10	10	10	10	10
mean ± SD	28.4 ± 2.7 **	30.4 ± 4.7	31.8 ± 5.0 **	29.7 ± 5.4 *	35.5 ± 8.0
n =number of animals examined SD: standard deviation * = p < 0.05, ** = p < 0.01 statistical significance

Table 2: Absolute and relative organ weights after 90 days of treatment

	Dose level (mg/kg bw/day)
Organ	Males				Females
	0	100	300	1000	0	100	300	1000
Liver weight
absolute (g)	14.10 ± 1.84	14.63 ± 1.96	14.06 ± 1.24	16.22 ± 1.35*	6.87 ± 0.89	7.10 ± 0.60	7.15 ± 0.70	7.62 ± 0.60
relative to bw (g/100g bw)	2.71 ± 0.31	2.71 ± 0.23	2.77 ± 0.19	3.02 ± 0.19*	2.53 ± 0.16	2.62 ± 0.09	2.73 ± 0.18*	2.81 ± 0.16**
Kidney weight
absolute (g)	3.05 ± 0.32	3.18 ± 0.31	3.25 ± 0.31	3.41 ± 0.38*	1.77 ± 0.14	1.83 ± 0.19	1.78 ± 0.12	1.92 ± 0.15
relative to bw (g/100g bw)	0.59 ± 0.07	0.59 ± 0.03	0.64 ± 0.07	0.64 ± 0.07	0.66 ± 0.05	0.67 ± 0.07	0.68 ± 0.04	0.71 ± 0.05

 * = p < 0.05 statistical significance

Table 3: Absolute and relative organ weight after 90 days of treatment followed by a 28-day recovery period

Organ	Males		Females
	0	1000	0	1000
Liver weight
absolute (g)	13.45 ± 1.21	15.35 ± 1.89 *	7.60 ± 0.79	7.09 ± 0.67
relative to bw (g/100g bw)	2.47 ± 0.09	2.71 ± 0.17 **	2.48 ± 0.23	2.42 ± 0.19
Spleen weight
absolute (g)	0.77 ± 0.07	0.90 ± 0.14 *	0.60 ± 0.08	0.60 ± 0.12
relative to bw (g/100g bw)	0.14 ± 0.01	0.16 ± 0.02 *	0.20 ± 0.02	0.20 ± 0.03
Lung weight
absolute (g)	1.46 ± 0.08	1.58 ± 0.15 *	1.13 ± 0.06	1.12 ± 0.05
relative to bw (g/100g bw)	0.27 ± 0.07	0.28 ± 0.01	0.37 ± 0.03	0.38 ± 0.05
Heart weight
absolute (g)	1.56 ± 0.16	1.74 ± 0.23	1.09 ± 0.11	0.94 ± 0.08 **
relative to bw (g/100g bw)	0.29 ± 0.02	0.31 ± 0.04	0.36 ± 0.04	0.32 ± 0.03 *
Thyroid weight
absolute (g)	25.6 ± 3.3	31.6 ± 4.9 **	17.8 ± 3.2	19.6 ± 3.5
relative to bw (g/100g bw)	4.7 ± 0.6	5.6 ± 0.9 *	5.8 ± 1.1	6.8 ± 1.6

 * = p < 0.05, ** = p < 0.01 statistical significance

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the NOAEL for systemic toxicity was 1000 mg/kg bw/day in male and female rats.