4-methoxybenzyl alcohol

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

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 form Study report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Principles of method if other than guideline:

To determine the toxicity likely to arise from the repeated exposure of the test item over a relatively limited period of time (28 days) in male and female SD rats.

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): 158.19g/mol
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations):Impurity : 1.34 %

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Details on test animal
TEST ANIMALS
- 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), ad libitum.

- 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

Vehicle:

other: Groundnut oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: 125, 250 and 500 mg was weighed and dissolved in 10 ml of groundnut oil by vortexing the resultant (concentration / dose).

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 and 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

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Doses were analyzed by using HPLC system.

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:

Details on study design
- Dose selection rationale: No data available
- Rationale for animal assignment (if not random): Animals were randomized by body weight and sex.
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Examinations

Observations and examinations performed and frequency:

Observations and examinations performed & frequency

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily
- Cage side observations performed: Visual inspection

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily for to characterize the onset and duration of clinical signs. However, detailed physical and clinical observations were conducted before the first exposure of test item and at least once a week thereafter till the end of the study.

BODY WEIGHT: Yes
- Time schedule for examinations: On day 1, 8, 15, 22, 28 and on day 29 before sacrifice.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

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

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Once weekly (on days 2, 9, 16 & 23)

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Fourth week of chemical treatment.
- Dose groups that were examined: All 56 animals were examined.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: On completion of 28 days of treatment and prior to necropsy.
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes, overnight fasting.
- How many animals: All 56 animals were examined.
- Parameters examined: Haemoglobin (Hb) , RBC Count, Total and differential leucocyte count, Haematocrit (Hct /PCV), Mean corpuscular volume (MCV), Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC) and Platelet Count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On completion of 28 days of treatment and prior to necropsy.
- Animals fasted: Yes, overnight fasting.
- How many animals: All 56 animals were examined.
- Parameters examined: Sodium, Potassium, Glucose, Total Cholesterol, Blood Urea, Creatinine, Total Protein Albumin, SGPT (Serum glutamic pyruvic transaminase)/ALT, SGOT (Serum glutamic oxaloacetic transaminase)/AST, Hormones analysis (testosterone and estrogen) and Total Bile acids.

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters examined: No data available

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Yes, Locomotor activity was observed.

OTHER:
Organ weight were measured.

Sacrifice and pathology:

Sacrifice and pathology
GROSS PATHOLOGY: Yes
On completion of treatment, all surviving rats were sacrificed and a complete necropsy was carried out on all animals. Tissues were collected from brain, stomach, large and small intestine, liver, kidneys, adrenal gland(s), spleen, heart, thymus, lungs, testis/ovary, uterus, lymph nodes, peripheral nerve (sciatic), bone marrow, and other gross lesions, if any.
Tissues were preserved in 10% formal saline. However testes, ovaries and uterus were first fixed in Bouin’s fixative for two hours then transferred to 10% formal saline.

HISTOPATHOLOGY: Yes
Histological examination was conducted on tissues/organs from the control and the high-dose group animals Organ examined: lung, liver, kidney, heart, spleen, testis, ovaries, adrenal glands, large and small intestine, brain, sciatic nerve, lymph nodes, bone marrow, stomach and thymus were examined.

Other examinations:

The collected organs were also weighed.

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.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

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

Mortality:

no mortality observed

Description (incidence):

All animals except animal No. 25 & 26 of high dose survived throughout the treatment period.

Body weight and weight changes:

no effects observed

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 was observed in opthalmoscopic examination.

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):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

The weight of spleen was decreased in the 250 mg/kg body weight/day groups while treatment with 500 mg/kg body weight/day resulted in the decreased weight of kidney in both female and male rats. However, the absolute organ weight of male animals does not show any significant difference.

In male rats, the relative weight of liver was noticed to be increased at 125 mg/kg body weight/day. Significant increase in the relative weights of heart and testes was also noticed at 500 mg/kg body weight/day.

In female rats, at all dose levels, the relative weight of ovaries was decreased in comparison to the control group animals. At 250 mg/kg body weight/day, the relative weights of brain and heart were significantly decreased. The relative weight of uterus was also decreased in the 125 or 250 mg/kg body weight/day groups.A significant decrease in the relative weight of spleen was observed at250 or 500 mg/kg body weight/day.

The absolute organ weight of ovaries from all treated groups was found to be decreased significantly in female rats.

The weight of uterus was found to be significantly decreased in 250 or 500 mg/kg body weight/day-treated male and female rats.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Nasal discharge, red crust around nostril, perineum wet, bloated stomach, viscous oily secretion was found inside the thoracic cavity.

Small white solid mass slightly firm in consistency floating inside the urinary bladder was observed.

However, due to adaptive metabolic and physiological changes, anoxic/ hypoxic conditions during anesthesia and terminal sacrifice, these findings were considered to be of no toxicological significance.

Neuropathological findings:

not specified

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

Effect levels

Dose descriptor:

NOAEL

Effect level:

125 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

ophthalmological examination

organ weights and organ / body weight ratios

water consumption and compound intake

other: No effect observed

Target system / organ toxicity

Critical effects observed:

not specified

System:

other: not specified

Organ:

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 when Sprague Dawley male and female rats were exposed daily to test substance by oral route for 28 days.

Executive summary:

In a 28 days repeated dose toxicity study, the effect of test substance 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. 2 animals died at 500 mg/kg bw as compared to control.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. A significant decrease in body weights in male rats on the first, second and fourth week of treatment and in female rats, a significant decrease in body weight was observed on the first, second and third week of treatment were observed as compared to control. No effects on food and water consumption were observed as compared to control. In addition,In hematology, methyl 2-naphthyl ether resulted in significantly increased platelet count along with decreased MCV, MCH and MCHC levels. Treatment with 250 mg/kg body weight/day resulted in significantly decreased levels of hemoglobin and MCHC in male rats. At 250 mg/kg body weight/day, a significant decrease was noticed in neutrophils along with altered values of MCV in female rats. In clinical chemistry, significantly increased the level of testosterone in the 500 mg/kg body weight/day group in males, as well as it significantly increased the level of estrogen in the 250 mg/kg body weight/day group in females. Significant increased levels in cholesterol, potassium and albumin was also observed. Similarly, Changes in relative and absolute organ weight of spleen, kidney, liver, brain, heart, ovaries and uterus were observed when treated with 125, 250 or 500 mg/kg body weight/day. Nasal discharge, red crust around nostril, perineum wet, bloated stomach, viscous oily secretion was found inside the thoracic cavity. Small white solid mass slightly firm in consistency floating inside the urinary bladder was observed. However, due to adaptive metabolic and physiological changes, anoxic/ hypoxic conditions during anesthesia and terminal sacrifice, these findings were considered to be of no toxicological significance. 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. Since the histopathological changes were not observed in 125 mg/kg bw and these findings were also observed in the control animals. Hence, NOAEL was considered to be 125 mg/kg bw when Sprague Dawley male and female rats were exposed daily to test substance by oral route for 28 days.