1,3-dihydro-5-methoxy-2H-benzimidazole-2-thione

Toxicity to reproduction

Administrative data

Endpoint:

toxicity to reproduction

Remarks:

other: subacute repeated dose study

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from study report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 407

Principles of method if other than guideline:

Repeated dose 28- days oral toxicity study of test chemical was performed in Sprague Dawley (SD) rats to assess the reproductive toxicity.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Methyl 2-naphthyl ether

EC Number:

202-213-6

EC Name:

Methyl 2-naphthyl ether

Cas Number:

93-04-9

Molecular formula:

C11H10O

IUPAC Name:

2-methoxynaphthalene

Details on test material:

- Name of test material
(as cited in study report): Methyl 2-naphthyl ether (MNE) (Batch no. 0001)
- Molecular formula
(if other than submission substance): C11H10O
- Molecular weight
(if other than submission substance): 158.19999999999999 g/mol
-Substancetype: Organic
-Physicalstate: Solid
-Impurities (identity and concentrations): Impurity : 1.34 %

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

No Data Available

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Central Animal Facility (CAF), NIPER, Sector-67, S.A.S. Nagar, Punjab, India
- Age at study initiation: 7 to 8 weeks old
- Weight at study initiation: Male 184.38-234.56 g, Female 176.90-208.56 g
- Fasting period before study: No data available
- Housing: Four rats per sex per cage were housed in sterilized solid bottom polypropylene cages with stainless steel grill tops with bedding of clean paddy husk. The cages were suspended on stainless steel racks in a controlled environment.
- Diet (e.g. ad libitum): Standard laboratory sterile extruded pelleted rodent feed (Provimi Animal Nutrition India Pvt. Ltd, Bangalore, India), ad libitum
- Water (e.g. ad libitum): Potable tap water filtered through Reviva Reverse Osmosis System (water filter cum purifier), adlibitum
- Acclimatization period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%):30-70 %
- Air changes (per hr): 25 ± 5 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: 11.04.2014 (Male), 13.04.2014 (Female); To: 17.05.2014 (Male), 19.05.2014 (Female)

Administration / exposure

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Dissolved the chemical in Groundnut oil.

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Groundnut oil
- Concentration in vehicle: 0, 125, 250 or 500 mg/kg body weight/day
- Amount of vehicle (if gavage): 5 ml/kg body weight
- Lot/batch no. (if required): No data available
- Purity: No data available 98.66%

Details on mating procedure:

- M/F ratio per cage: No data available
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: No data available
- After successful mating each pregnant female was caged (how): No data available
- Any other deviations from standard protocol: No data available

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No Data Available

Duration of treatment / exposure:

28 days

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

Doses / concentrations 4

Dose / conc.:

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

No. of animals per sex per dose:

Total animals: 56
Control: 7 males, 7 females
125 mg/kg/day: 7males, 7 females
250 mg/kg/day: 7 males, 7 females
500 mg/kg/day: 7 males, 7 females

Control animals:

yes, concurrent vehicle

Details on study design:

No Data Available

Positive control:

No Data Available

Examinations

Parental animals: Observations and examinations:

The mortality and changes in body weight, food consumption and water consumption were recorded.

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

No data available

Postmortem examinations (parental animals):

The testes, ovaries and uterus were pathologically examined after termination. Examinations also included weight of brain, testes and ovaries (incl. paired ovaries and uterus, including cervix).

Postmortem examinations (offspring):

No data available

Statistics:

Statistical analysis was carried out by using Microsoft Excel and IBM SPSS statistics version -20. All analyses and comparisons were evaluated at the 5 % level, statistically significant differences (p0.05) indicated by appropriate notation. PAIRED T-Testing procedure was used to check the significance between above mentioned groups. For multiple comparisons Turkey’s HSD test was applied.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Abnormalities such as nasal discharge, red crust around the nostrils was observed in all treatment groups. In addition, a few cases of snuffling, eye lid swelling and hunched back posture were also observed in the treated groups.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, non-treatment-related

Description (incidence):

All animals except two animals of high dose group survived throughout the treatment period.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Treatment with 500 mg/kg body weight/day showed a significant reduction in body weights in male rats on the first, second and fourth week of treatment. In female rats, a significant decrease in body weight was observed on the first, second and third week of treatment.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No difference was observed among the groups.

Food efficiency:

not specified

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

no effects observed

Description (incidence and severity):

No difference in water consumption was observed among the groups.

Ophthalmological findings:

no effects observed

Description (incidence and severity):

No difference in opthalmological changes was observed among the groups.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

In male rats, significant increase in the platelet count was observed along with decrease in MCV and MCH levels at 125 mg/kg body weight/day.
Treatment with 250 mg/kg body weight/day showed significantly decreased values of hemoglobin and MCHC in male rats. The level of MCHC was also found to be significantly decreased when treated with 500 mg/kg body weight/day.

In female rats, treatment with 125 mg/kg body weight/day showed significant alterations in platelet count and MCV. At 250 mg/kg body weight/day, a significant decrease was noticed in neutrophils along with altered values of MCV in female rats.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

In female animals, the level of total bile acid was decreased at 250 mg/kg body weight/day. A significant increase was noticed in the level of estrogen in the same group. A similar trend was also noticed at 125 and 500 mg/kg body weight/day, but was statistically non-significant.

At 250 and 500 mg/kg body weight/day the levels of potassium and albumin were found to be significantly increased. The cholesterol level in the 250 mg/kg body weight/day group was noticed to be significantly elevated in the mid-dose group.

In male rats, the level of testosterone was significantly increased after treatment with 500 mg/kg body weight/day.

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No statistically significant difference was observed in the motor activity scores for any dose group when compared with the control animals.

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No remarkable changes observed in control and the 500 mg/kg body weight/day-treated animals.

A few microscopic findings observed in 500 mg/kg body weight/day-treated animals included collapsed lung with focal inflammation, focal fatty change and excess of lymphocytes in liver, inflammation in small intestine, reactive spleen and excess of mucous in colon. However, these findings were also observed in the control animals.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Effect levels (P0)

1

Dose descriptor:

NOAEL

Effect level:

125 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

water consumption and compound intake

ophthalmological examination

haematology

clinical biochemistry

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

other: No effect observed

2

Dose descriptor:

LOAEL

Effect level:

250 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

organ weights and organ / body weight ratios

other: effect observed on reproductive organs

Target system / organ toxicity (P0)

Critical effects observed:

yes

Lowest effective dose / conc.:

125 mg/kg bw/day (nominal)

System:

female reproductive system

Organ:

ovary

Treatment related:

yes

Dose response relationship:

not specified

Relevant for humans:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

other: not specified

Generation:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

not measured/tested

Target system / organ toxicity (F1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Any other information on results incl. tables

SUMMARY OF BODY WEIGHTS(g)

Male

Group   Dose mg/kg       Day1     Day8     Day15     Day22     Day28   Terminal Day     1     Vehicle Mean 211.80 242.34 262.97 283.01 300.76 296.93 S.D. 14.03 14.91 16.29 24.57 26.41 27.19   S.E.M. 6.62 6.15 6.20 8.68 8.78 9.16     2     125 Mean 218.02 243.85 260.27 279.35 296.61 291.98 S.D. 9.56 11.42 11.59 14.54 14.88 14.86   S.E.M. 4.38 4.68 4.45 5.21 5.02 5.09     3     250 Mean 210.57 221.01 241.60 267.42 286.08 282.17 S.D. 9.82 26.98 30.11 23.54 25.05 24.52   S.E.M. 4.67 12.21 12.46 8.80 8.76 8.69     4     500 Mean 214.35 212.06 223.41 240.97 256.81 256.24 S.D. 10.64 10.08 11.64 13.72 6.86 9.27   S.E.M. 4.96 4.76 5.21 5.70      2.67 3.62

SUMMARY OF BODYWEIGHTS(g)

Female

Group	Dose mg/kg		Day1	Day8	Day15	Day22	Day28	Terminal Day
1	Vehicle	Mean	191.17	200.82	208.91	215.94	227.15	219.85
		S.D.	7.12	12.39	16.56	15.30	17.64	19.32
		S.E.M.	3.72	6.17	7.93	7.09	7.77	8.79
2	125	Mean	192.17	201.14	208.56	214.38	220.79	216.51
		S.D.	9.31	10.16	8.58	7.62	10.76	10.09
		S.E.M.	4.85	5.05	4.11	3.55	4.87	4.66
3	250	Mean	194.94	202.08	209.12	216.24	228.85	226.38
		S.D.	9.45	8.89	8.24	7.85	7.51	8.75
		S.E.M.	4.85	4.40	3.94	3.63	3.28	3.87
4	500	Mean	190.49	182.52	192.46	205.74	216.14	212.37
		S.D.	8.82	9.74	12.73	11.46	11.61	12.10
		S.E.M.	4.63	5.34	6.61	5.57	5.37	5.70

SUMMARY OF HEMATOLOGY DATA Male

Group/Dose   WBC 3 (x10/ mm3) RBC 6 (x10/ mm3) PLT (x10/mm3) Hct (%) Hgb (g/dl) MCV(fl) MCH (pg) MCHC(g/dl) Lym % Mon % Nut % Eos %   Bas%     1/0   Mean 12.06 69.49 4.34 23.27 1.17 0.99 10.15 52.77 53.54 16.40 31.07 16.67 600.2 9 ±S.D. 2.24 9.56 1.11 9.04 1.07 0.33 0.63 1.40 2.92 0.51 0.97 1.00 76.78 ± S.E.M. 18.59 13.76 25.46 38.85 91.51 33.35 6.17 2.65 5.45 3.11 3.12 5.99 12.79       2/125mg/kg   Mean 12.09 69.73 4.57 22.04 2.19 0.71 10.16 50.24＊ 51.00 15.49＊ 30.83 15.73 1118. 00＊   ±S.D. 2.18 5.30 0.71 6.81 2.70 0.17 0.53 1.93 2.36 0.71 0.83 0.80 383.1 9 ± S.E.M. 0.8 2.0 0.3 2.6 1.0 0.1 0.2 0.7 0.9 0.3 0.3 0.3 144.8       3/250mg/kg   Mean 14.97 63.19 4.66 28.04 2.36 0.93 9.93 52.76 52.41 15.81 29.99 ＊ 15.70 614.8 6   ±S.D. 2.34 11.26 0.75 12.16 2.83 0.20 0.24 2.50 3.03 0.80 0.18 0.96 173.0 5 ± S.E.M. 0.9 4.3 0.3 4.6 1.1 0.1 0.1 0.9 1.1 0.3 0.1 0.4 65.4       4/ 500mg/kg   Mean 9.10 53.17 3.28 16.91 2.29 0.60 7.32 38.41 39.25 11.61 21.62 ＊ 11.87 476.4 9   ±S.D. 5.76 31.42 2.26 6.52 1.66 0.35 4.89 25.12 25.52 7.57 14.70 7.70 295.4 8 ± S.E.M. 2.6 14.1 1.0 2.9 0.7 0.2 2.2 11.2 11.4 3.4 6.6 3.4 132.1

SUMMARY OF HEMATOLOGY DATA Female

emaleGroup/Dose			WBC (x103/mm3)	RBC (x106/mm3)	PLT (x103/mm3)	Hct (%)	Hgb (g/dl)	MCV(fl)	MCH (pg)	MCHC (g/dl)	Lym %	Mon %		Nut %	Eos %		Bas%
Vehicle	Mean		11.66	62.20	4.41	30.31	1.39	0.87	10.18	49.90	50.74	15.44		30.97	15.71		994.57
	±S.D.		2.19	8.62	1.09	8.69	1.61	0.17	0.55	2.34	2.56	0.68		0.31	0.84		520.25
	± S..E..M.		0.8	3.3	0.4	3.3	0.6	0.1	0.2	0.9	1.0	0.3		0.1	0.3		196.6
2      / 125mg/kg		Mean	12.28	67.97	4.34	25.10	0.94	0.90	10.13	51.66＊	52.26		15.86	30.71		16.06		518.71 ＊
		±S.D.	4.51	6.98	0.78	6.78	0.47	0.18	0.47	1.36	1.39		0.73	0.71		0.45		114.08
		±S.E.M.	1.7	2.6	0.3	2.6	0.2	0.1	0.2	0.5	0.5		0.3	0.3		0.2		43.1
3    / 250mg/kg		Mean	11.94	69.24	4.13	21.83＊	2.93	1.07	10.18	51.76＊	52.76		15.71	30.37	15.99			585.00
		±S.D.	3.21	7.96	0.54	7.77	2.71	0.24	0.45	1.55	3.36		0.75	1.44	1.17			213.78
		±S.E.M.	1.2	3.0	0.2	2.9	1.0	0.1	0.2	0.6	1.3		0.3	0.5	0.4			80.8
4       / 500mg/kg		Mean	11.37	66.26	4.36	26.57	1.06	1.03	10.00	51.77	51.79		15.79	30.47	15.79			614.29
		±S.D.	2.10	9.91	0.99	9.88	1.34	0.19	0.73	0.85	3.90		0.57	0.92	1.41			139.13
		±S.E .M.	0.8	3.7	0.4	3.7	0.5	0.1	0.3	0.3	1.5		0.2	0.3	0.5			52.6

SUMMARY OF CLINICAL BIOCHEMISTRY DATA Male Rats

Group/Dose		Sod.(mmol/L)		Pot.(mmol/L)		ALB(g%)		CHOL(mg%)		CRT(mg%)		SGOT(IU/L)	SGPT (IU/L)	GLU(mg%)	TP(g%)	BUN(mg%)	TBA	Test
																	ng/L	nmol/L
1   /  0	Mean	131.5		7.0		3.9		83.9		0.9		33.1	32.9	117.0	6.5	12.5	490.1	4.363
	±S.D.	8.6		0.5		0.3		18.1		0.2		4.0	5.1	7.1	0.4	3.0	44.82	1.02
	±S.E.M.	3.2		0.2		0.1		6.8		0.1		1.5	1.9	2.7	0.1	1.1	16.94	0.387
2  /125 mg/kg	Mean		130.0	7.6	3.7		87.6		0.9		33.9		32.5	119.1	6.5	14.4	485.4	5.421
	±S.D.		3.3	0.7	0.3		10.7		0.2		8.2		7.6	4.9	0.5	3.0	36.17	1.03
	±S.E.M.		1.2	0.3	0.1		4.0		0.1		3.1		2.9	1.9	0.2	1.1	13.67	0.392
3 /250 mg/kg	Mean		130.1	7.1	3.7		87.7		0.9		65.4		30.0	121.0	6.7	12.1	465.4	4.716
	±S.D.		9.8	1.0	0.2		17.4		0.1		90.0		7.0	9.5	0.5	2.8	35.10	1.28
	±S.E.M.		3.7	0.4	0.1		6.6		0.1		34.0		2.6	3.6	0.2	1.1	13.26	0.487
4 / 500 mg/kg	Mean		124.5	11.0	3.7		96.2		0.9		31.1		30.5	121.6	6.4	13.7	485.2 *	6.289*
	±S.D.		11.1	8.4	0.3		12.1		0.2		7.5		6.4	13.8	0.5	4.0	23.33	0.398
	±S.E.M.		5.0	3.8	0.1		5.4		0.1		3.3		2.8	6.2	0.2	1.8	10.43	0.178

SUMMARY OF CLINICAL BIOCHEMISTRY DATA - Female

Group /Dose		Sod.	Pot.	ALB	CHOL	CRT	SGOT	SGPT	GLU	TP	BUN	TBA	Est
		(mmol/L)	(mmol/L)	(g%)	(mg%)	(mg%)	(IU/L)	(IU/L)	(mg%)	(g%)	(mg%)	ng/L	ng/L
1        / 0 mg/kg	Mean	139.2	7.2	3.3	93.0	1.1	32.3	39.9	110.4	7.0	12.9	485	9.156
	±S.D.	9.3	0.6	0.3	14.5	0.2	15.3	21.7	14.2	0.5	4.3	30.60	1.47
	±S.E.M.	3.5	0.2	0.1	5.5	0.1	5.8	8.2	5.4	0.2	1.6	11.56	0.558
2     / 125 mg/kg	Mean	136.6	7.8	3.4	98.0	1.1	28.8	39.6	100.0	7.3	12.5	475.1	9.869
	±S.D.	4.3	0.9	0.1	17.1	0.4	7.4	9.0	8.9	0.5	2.1	30.59	1.27
	±S.E.M.	1.6	0.3	0.1	6.5	0.1	2.8	3.4	3.4	0.2	0.8	11.56	0.482
3     / 250 mg/kg	Mean	141.6	8.1	3.8	119.9	1.1	27.9	40.0	100.2	7.0	12.5	425.5*	10.478*
	±S.D.	7.3	0.9	0.4	24.7	0.2	6.0	8.1	9.0	0.6	2.8	52.38	1.33
	±S.E.M.	2.8	0.4	0.2	9.3	0.1	2.3	3.0	3.4	0.2	1.1	19.79	0.505
4     / 500 mg/kg	Mean	137.0	8.1	3.7	113.4	1.1	32.3	43.5	100.1	7.0	14.1	448.5	9.842
	±S.D.	5.9	0.8	0.2	23.6	0.2	8.4	9.8	5.8	0.6	2.3	31.16	1.93
	±S.E.M.	2.2	0.3	0.1	8.9	0.1	3.2	3.7	2.2	0.2	0.9	13.93	0.73

SUMMARY OF ABSOLUTE ORGAN WEIGHTS(g) - Male Rats

Group/ Dose		Brain	Thymus	Liver	Kidneys	Adrenals	Ovaries	Spleen	Heart	Epididymides
1 Vehicle	Mean	1.79	0.30	10.72	1.98	0.03	2.79	1.04	0.55	0.88
	±S.D.	0.28	0.06	1.48	0.30	0.01	0.27	0.15	0.11	0.09
	±S.E.M.	0.11	0.02	0.56	0.11	0.01	0.10	0.05	0.04	0.03
2 (125) mg/kg	Mean	1.89	0.46	11.54	2.09	0.07	2.75	1.05	0.54	0.89
	±S.D.	0.12	0.16	0.70	0.11	0.08	0.22	0.06	0.13	0.08
	±S.E.M.	0.05	0.06	0.26	0.04	0.03	0.08	0.02	0.05	0.03
3 (250)mg/kg	Mean	1.87	0.32	11.25	2.04	0.04	2.81	1.10	0.63	0.91
	±S.D.	0.07	0.06	1.96	0.23	0.01	0.14	0.12	0.19	0.10
	±S.E.M.	0.02	0.02	0.74	0.09	0.00	0.05	0.05	0.07	0.04
4  (500) mg/kg	Mean	1.79	0.28	10.61	1.93	0.04	2.87	1.03	0.54	0.88
	±S.D.	0.07	0.05	0.51	0.17	0.01	0.13	0.06	0.11	0.05
	±S.E.M.	0.03	0.02	0.19	0.06	0.00	0.05	0.02	0.04	0.02

 SUMMARY OF ABSOLUTE ORGAN WEIGHTS(g)

 Femalerats

Group/ Dose		Brain	Thymus		Liver		Kidneys		Adrenals		Ovaries	Spleen	Heart		Uterus
1/0    (mg/kg)	Mean	1.79639	0.69686		8.13916		1.66367		0.0451		0.14289	0.63041	0.79684		0.46711
	±S.D.	0.13458	0.547		0.47047		0.18627		0.01172		0.02302	0.11848	0.11455		0.07792
	±S.E.M.	0.05087	0.20675		0.17782		0.0704		0.00443		0.0087	0.04478	0.04329		0.02945
2/(120) mg/kg	Mean	1.78147	0.28884	8.15586		1.55246		0.04734			0.08319＊	0.63869	0.77013		0.39303
	±S.D.	0.11628	0.1304	1.06437		0.16655		0.00937			0.01815	0.24329	0.06459		0.08246
	±S.E.M.	0.04395	0.04929	0.40229		0.06295		0.00354			0.00686	0.09196	0.02441		0.03117
2/   250 mg/kg	Mean	1.6069	0.3289	8.3645		1.566		0.0395			0.0935＊	0.4718	0.7185		0.4667＊
	±S.D.	0.27569	0.12711	0.78925		0.15522		0.01286			0.02423	0.11176	0.06134		0.07708
	±S.E.M.	0.1042	0.04804	0.29831		0.05867		0.00486			0.00916	0.04224	0.02319		0.02913
4  / (500) mg/kg	Mean	1.75207	0.42151	7.9445		1.525＊		0.04407		0.07481		0.49059＊		0.70961	0.34757＊
	±S.D.	0.12692	0.20921	0.63133		0.13336		0.00665		0.01518		0.09178		0.02657	0.10007
	±S.E.M.	0.04797	0.07907	0.23862		0.05041		0.00251		0.00574		0.03469		0.01004	0.03782

SUMMARY OF RELATIVE ORGAN WEIGHTS Male Rats

Group/Dose		Brain	Thymus	Liver	Kidneys	Adrenals	Testes	Splee n	Heart	epididymide s
1/   0	Mean	0.6051	0.1009	3.6170	0.6680	0.0110	0.9447	0.1846	0.2970	0.3521
	±S.D.	0.1132	0.0236	0.4211	0.0880	0.0052	0.1114	0.0407	0.0210	0.0527
	±S.E.M.	0.0428	0.0089	0.1592	0.0333	0.0020	0.0421	0.0154	0.0079	0.0199
2/ 125 mg/kg	Mean	0.6466	0.1549	3.9569*	0.7236	0.0243	0.9377	0.1856	0.3060	0.3599
	±S.D.	0.0330	0.0515	0.2711	0.0369	0.0274	0.0434	0.0428	0.0262	0.0299
	±S.E.M.	0.0125	0.0195	0.1025	0.0139	0.0104	0.0164	0.0162	0.0099	0.0113
3  / 250 mg/kg	Mean	0.6674	0.1140	3.9886	0.7207	0.0143	1.0007	0.2209	0.3220	0.3883
	±S.D.	0.0627	0.0252	0.6023	0.0431	0.0021	0.0816	0.0608	0.0367	0.0346
	±S.E.M.	0.0237	0.0095	0.2277	0.0163	0.0008	0.0309	0.0230	0.0139	0.0131
4   / 500 mg/kg	Mean	0.5317	0.0868	3.1877	0.5684	0.0110	0.8401	0.1709	0.2618*	0.3071*
	±S.D.	0.3033	0.0461	1.7161	0.3359	0.0062	0.4873	0.0898	0.1468	0.1775
	±S.E.M.	0.1357	0.0206	0.7675	0.1502	0.0028	0.2179	0.0402	0.0656	0.0794

SUMMARY OF RELATIVE ORGAN WEIGHTS- Females

Group/Dose		Brain	Thymu s	Liver	Kidneys	Adrenal s		Ovaries		uterus	Spleen	Heart
1/0	Mean	0.8181	0.3176	3.7199	0.7573	0.0200		0.0654		0.2107	0.2849	0.3614
	±S.D.	0.0380	0.2566	0.3170	0.0667	0.0045		0.0141		0.0202	0.0354	0.0334
	±S.E.M.	0.0144	0.0970	0.1198	0.0252	0.0017		0.0053		0.0076	0.0134	0.0126
2 / 125 mg/kg	Mean	0.8227	0.1324	3.7541	113.7454		0.0214		*0.0380	0.1824	0.2926	0.3557
	±S.D.	0.0439	0.0598	0.3280	299.0822		0.0053		0.0090	0.0439	0.1052	0.0323
	±S.E.M.	0.0166	0.0226	0.1240	113.0424		0.0020		0.0034	0.0166	0.0398	0.0122
3  /          250 mg/kg	Mean	0.7137	0.1443	3.6900	0.6917	0.0171		0.0406*		0.2054	0.2077*	0.3169*
	±S.D.	0.1425	0.0553	0.2348	0.0629	0.0053		0.0089		0.0298	0.0487	0.0212
	±S.E.M.	0.0539	0.0209	0.0887	0.0238	0.0020		0.0034		0.0113	0.0184	0.0080
4    /  500mg/kg	Mean	0.8253	0.1970	3.7417	0.7173	0.0203		0.0349*		0.1636	0.2304*	0.3341
	±S.D.	0.0546	0.0920	0.2484	0.0423	0.0028		0.0080		0.0463	0.0405	0.0130
	±S.E.M.	0.0206	0.0348	0.0939	0.0160	0.0011		0.0030		0.0175	0.0153	0.0049

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 125 mg/kg bw for male rats and LOAEL was considered to be 250 mg/kg body weight/day for female when Sprague Dawley male and female rats were exposed daily to the test chemical by oral route for 28 days.

Executive summary:

In a 28 days repeated dose toxicity study, the effect of the test chemical was evaluated in male and female Sprague Dawley rats. The test chemical was administered by oral gavage in the concentration of 0, 125, 250 or 500 mg/kg body weight/day. The results showed that the test chemical significantly increased the level of testosterone in the 500 mg/kg body weight/day group as well as it significantly increased the level of estrogen in the 250 mg/kg body weight/day group .The relative and absolute organ weight of ovaries decreased when treated with 125, 250 or 500 mg/kg body weight/day. In similarity, the relative and absolute organ weight of uterus decreased in the 125 or 500 mg/kg body weight/day groups.No significant changes in were detected in hematology,clinical biochemistry,mortality ororgan weight, and no effects were observed in water consumption,opthalmoscopic examination or locomotor activity. In male rats, the relative organ weights of the testes and epididymides increased when rats were treated with 500 mg/kg body weight/day. Histopathology performed on reproductive organs after treatment with 500 mg/kg body weight/day did not reveal any toxic lesions as compared to control. Hence, NOAEL was considered to be 250 mg/kg bw for male rats and LOAEL was considered to be 125 mg/kg body weight/day for female when Sprague Dawley male and female rats were exposed daily to the test chemical by oral route for 28 days.