Reaction products of diphenylamine with nonene, branched

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Remarks:

ten weeks premating administration period, estrous cycle determination performed at the end of the premating administration period (in a three weeks period before the start of mating), recovery groups for high and control group

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Justification for study design:

10 weeks premating was requested by ECHA for the analogue substance for the e1Gen (OECD 443) study. This study was set up as the bridging study for this endpoint (COLLA).

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of Benzeneamine, N-phenyl- with nonene (branched)

EC Number:

701-385-4

Cas Number:

36878-20-3

Molecular formula:

C21H29N - C30H47N (main constituents)

IUPAC Name:

Reaction products of Benzeneamine, N-phenyl- with nonene (branched)

Test material form:

liquid: viscous

Details on test material:

100 g/100 g (UVCB substance)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
BASF and 0016046440
- Expiration date of the lot/batch:
07 Jan 2020

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: ambient (room temperatures)
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None

- Preliminary purification step (if any): None

FORM AS APPLIED IN THE TEST: test substance addition to the diet

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl:Wl(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 35±1 d (arrival at test facility)
- Weight at study initiation: (P) Males: 116g; Females: 96 g
- Housing: group housed - up to 5 animals/cage (pretreatment), 2 animals/cage (premating), 2 male animals/cage (mating and postmating)
- Polysulfonate cages Typ 2000P (H-Temp)
- Diet: Mouse and rat maintenance diet "GLP", Granovit AG, Kaiseraugst, Switzerland; ad libitum
- Water: Drinking water ad libitum
- Acclimation period: 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 45 - 65
- Air changes (per hr): 15
- Photoperiod (hrs dar k / hrs light): 12/12

IN-LIFE DATES: From: 2019-05-21 To: 2019-09-25 (2019-10-10 recovery females)

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

PREPARATION OF DIET:
The required quantity of test substance was weighed in a beaker depending on the dose group and thoroughly mixed with a small amount of food. Then further amounts of food were
added to this premix and thoroughly mixed for 3 minutes. Afterwards, further amounts of food, depending on the dose group, were added to this premix in order to obtain the desired
concentrations.

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with food.
- Storage temperature of food:

The food used was mouse and rat maintenance diet “GLP”, meal, supplied by Granovit AG, Kaiseraugst, Switzerland.

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 (GD 0) of pregnancy
- After successful mating each pregnant female was caged: individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

At the beginning and towards the end of the premating phases, once during gestation and once during lactation of the Study each 3 samples were taken from the lowest and highest
concentration for potential homogeneity analyses. These samples were used as aconcentration control at the same time. At the above mentioned time points additionally one
sample from the mid concentration was taken for concentration control analysis.

The test item was a mixture of components. Thus, data evaluation was based on the integration of extracted ion chromatograms, being representative for the test item.
Furthermore, spiking experiments had shown that recovery of the test item was 91% or 107% for low (~750 mg/kg) or high (~3000 mg/kg) concentrations, respectively.
These recovery rates were in the range of the error of the method and therefore the content found was not corrected by the recovery rates. The concentration control was considered
as achieved when the mean recovery of a given sample was 90% ≤ x ≤ 110%. The samplewas considered as homogeneous when the standard deviation was RSD/MW ≤ 5%.
These requirements were fulfilled for all investigated samples and thus the concentration control as well as homogeneity was achieved

Duration of treatment / exposure:

12 weeks (males) (10 weeks premating+2 weeks mating)
16 weeks (females) (10 weeks premating+2 weeks mating + 3 weeks gestation + 13 days lactation)

Frequency of treatment:

daily for 7 days a week

Details on study schedule:

- Age at mating of the mated animals in the study: 15 weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

500 ppm

Remarks:

dose applied to animals during the premating and mating phase (males and females), postmating (males) and gestation phase and after weaning (females)

Doses / concentrations 2

Dose / conc.:

1 500 ppm

Remarks:

dose applied to animals during the premating and mating phase (males and females), postmating (males) and gestation phase and after weaning (females)

Doses / concentrations 3

Dose / conc.:

5 000 ppm

Remarks:

dose applied to animals during the premating and mating phase (males and females), postmating (males) and gestation phase and after weaning (females);
dose was also applied to recovery animals

Doses / concentrations 4

Dose / conc.:

250 ppm

Remarks:

dose applied to F0 females during lactation phase to maintain dams at desired target dose of the test substance: adjustment to historical body weight and food consumption data

Doses / concentrations 5

Dose / conc.:

750 ppm

Remarks:

dose applied to F0 females during lactation phase to maintain dams at desired target dose of the test substance: adjustment to historical body weight and food consumption data

Doses / concentrations 6

Dose / conc.:

2 500 ppm

Remarks:

dose applied to F0 females during lactation phase to maintain dams at desired target dose of the test substance: adjustment to historical body weight and food consumption data

No. of animals per sex per dose:

10
(plus 10 for control and high dose for recovery group)

Control animals:

yes

Details on study design:

- Dose selection rationale: requested by the sponsor
- Rationale for animal assignment: rats will be randomized according to their weight and allocated to the dose groups
- Fasting period before blood sampling for clinical biochemistry: 16-20 hours

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule for moribund and dead animals: twice daily (Mon-Fri), once daily (Sat, Sun and public holidays)
- Time schedule for clinical signs: at least once daily, if signs occur: several times daily

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: once a week for animals in the main group and the recovery group
- not determined in females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period
- during mating, females will be weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20
- females with litter will be weighed on the day after parturition (PND1) and on PND 4, 7, 10 and 13

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption will be determined once a week for the male and female parental animals of main groups as well as the males and females of recovery groups.
- not be determined during mating, in the females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period
- females with evidence of sperm: food consumption will be determined for GD 0-7, 7-14 and 14-20
- females which gave birth to a litter: food consumption will be determined for PND 1-4, 4-7, 7-10 and 10-13

OTHER:

Oestrous cyclicity (parental animals):

- in all parental females in the premating phase: estrous cycle length and normality was evaluated by preparing vaginal smears during a minimum of 3 weeks prior to mating and throughout cohabitation until there was evidence of sperm in the vaginal smear
- in all females of the recovery groups: estrous cycle length and normality will be evaluated in the 2 weeks of the recovery period
- in all parental females: estrous status was determined on the day of scheduled sacrifice

Sperm parameters (parental animals):

Parameters examined in F0 male parental generations and males of the recovery group:
- after organ weight determination, parameters determined in the right testis or right epididymis:
- Cauda epididymis sperm motility
- Sperm morphology
- Spermatid head count in the testis
- Sperm head count in the cauda epididymis
Parameters exmined for the control and highest dose group:
- Sperm morphology and sperm head count (cauda epididymis and testis)

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals after the end of the administration period the last litters in each generation were produced.
- Female animals: All surviving animals after the last litter of each generation was weaned (PND 14).
- Recovery animals: All surviving animals will be maintained for at least two weeks (recovery period) longer compared to the respective main groups.

GROSS NECROPSY
- Gross necropsy performed for all animals in the main and the recovery group
- consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera
- special attention being given to reproductive organs

HISTOPATHOLOGY / ORGAN WEIGHTS
Organs/Tissues weighed for...:
- all animals (main test and recovery group): anesthetized animals (final body weight), Epididymides, Ovaries, Prostate (ventral and dorsolateral part together, fixed), Seminal vesicles with coagulating glands (fixed), Testes, Thyroid glands (with parathyroid glands) (fixed), Uterus with cervix
- 5 animals/sex/test group (main test and recovery group): Adrenal glands (fixed), Brain, Heart, Kidneys, Liver, Spleen, Thymus (fixed)
Organs/Tissues fixed (4% neutral buffered formaldehyde) for...:
- all parental animals (main test and recovery group): All gross lesions, Adrenal glands, Aorta, Bone marrow (femur), Brain, Cecum, Cervix, Coagulating glands, Colon, Duodenum, Esophagus, Extraorbital lacrimal glands, Epididymides, left (modified Davidson's solution), Eyes with optic nerve (modified Davidson's solution), Femur with knee joint, Heart, Ileum, Jejunum (with Peyer's patches), Kidneys, Larynx

Postmortem examinations (offspring):

SACRIFICE
- One selected male and one female pup/litter were sacrificed at 13 days of age (PND 13) for blood sampling and thyroid/parathyroid glands fixation.
- All F1 offspring
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

Statistics:

Beside means and standard deviations, details statistics are given in table 1.

Offspring viability indices:

Pups Surviving days 0-4
Pups Surviving days 4-13

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Decreased body weight in males during in-life, including premating, from study day
21 to 91 with a maximum of -12.1% on study day 77 and -12.0% at the end of the
administration period in the main group
• Decreased body weight in females during premating from study day 28 to 70 with a
maximum of -11.5 % on study day 56 and -10.8% at the end of the administration
period in the main group
• Decreased body weight in recovery females during in-life phase from study day
21 to 119 with a maximum of -16.8 % (on study day 119) and during recovery
phase from study day 121 to 134 with a maximum of -15.2 % on study day 121 and
-10.6% at the end of the recovery period
• Decreased body weight in females from gestation day 0 to 20 with a maximum of
-18.5% on gestation day 20 and from lactation day 1 to 13 with a maximum of
-17.6% on lactation day 1 and resulting in -16.9% on lactation day 13
• Decreased body weight changes in males during in-life from study day 0 to 91
(-16.5%) and in females from study day 0 to 70 (-20%) in the main group

In the satellite male animals a decreased body weight gain was observed only during study day 56-63 and a non-statistically significant decrease from study day 0 to 91 (-7%) of the administration period followed by an increased body weight gain over the entire 14-day recovery period (+82%).
In the satellite (non pregnant) females a decreased body weight gain was observed during the administration period (-26%) and an increased body weight gain in the 14-day recovery period (+155%).

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Decreased food consumption in females from study day 7 onwards (up to -26.8% between study day 42 and 49) resulting in -11.6% from study day 0 to 70 in the main group
Decreased food consumption in satellite females during in-life phase (administration period) on study days 0 to 119 (-15.6%)
Decreased food consumption in females during gestation days 0 to 20 (-20.1%) and lactation days 1 to 13 (-23.5%)

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

On day 94 (males) Increase in ALT (1.03 vs 0.78 ukat/L) and ALP (2.28 vs 1.33 ukat/L) at the high dose group, reversible within 14-day recovery period
On day 113 (females): Increase in ALP ( 4.16 vs 0.91 ukat/L) and GGT_C (45 vs 25 nkat/L) at the high dose group, reversible within 14-day recovery period

Reduction in bile acids

Increase in triglycerides

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Description (incidence and severity):

No effects on thyroid hormones

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

effects observed, treatment-related

Description (incidence and severity):

Estrous cycle data, generated during the premating phase, revealed regular cycles in the females of all test groups including the control. The mean number of estrous cycle was 4.2 / 4.3 / 4.5 and 3.9 in test groups 0 - 3. The estrous cycle length in the different test groups was identical: 4.0 days in the test groups 0 – 2. In test group 3 (5000 ppm) the cycle length was 4.3 days which was not statistically significant different to the current control but outside of the historical control range 3.82-4.03 days.

All animals were cycling normally (3-6 days). No animal had a long estrous (3 days). One animal of the high dose group had a long distrous (4 days)

Reproductive function: sperm measures:

no effects observed

Description (incidence and severity):

After the administration period, regarding the incidence of abnormal sperms in the cauda epididymidis, sperm head counts in the testis and in the cauda epididymidis no treatmentrelated
effects were observed. The sperm motility in males of test group 1 (5000 ppm) was significantly lower compared to controls, but this change was not dose-dependent and
therefore it was regarded as incidental and not treatment-related.
After the recovery period, in males of test group 13 (5000 ppm) motility of the sperms and sperm head counts in the testis were significantly decreased. However, the values were
within historical control ranges (males, motility 79-93 %, sperm head counts in the testis 87- 126 Mio/g testis). Therefore, these alterations were regarded as incidental and not
treatment-related. Sperm head counts in the cauda epididymidis and incidences of abnormal sperms were not changed.

Effect levels (P0)

1

Dose descriptor:

NOEL

Effect level:

500 ppm (analytical)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

organ weights and organ / body weight ratios

histopathology: non-neoplastic

2

Dose descriptor:

NOAEL

Effect level:

1 500 ppm

Based on:

test mat.

Sex:

female

Basis for effect level:

reproductive function (oestrous cycle)

reproductive performance

Target system / organ toxicity (P0)

Critical effects observed:

yes

Lowest effective dose / conc.:

1 500 ppm

System:

hepatobiliary

Organ:

liver

Treatment related:

yes

Dose response relationship:

yes

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Viability index (days 0-4) groups 0 -3: 100 / 100 / 98.1 / 96.1
Viability index (days 4-13) groups 0-3: 100 / 100 / 100 / 100

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Offspring body weight (male and female) in the high dose group is lower than that of the control group starting day 7. On day 13, the mean body weight is 26.4g versus 32.5g (-18.8%)

Ophthalmological findings:

not examined

Haematological findings:

not examined

Urinalysis findings:

not examined

Anogenital distance (AGD):

no effects observed

Nipple retention in male pups:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

No changes in absolute organ weights in males (slight reduction in heart weight at the high dose group)
Reduction in absolute weight of ovaries (Control 111 mg, LD 104 mg, MD 91 mg and HD 67 mg)

Relative organ weights returned to normal within the 14-day recovery period.

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 500 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Target system / organ toxicity (F1)

Critical effects observed:

yes

Lowest effective dose / conc.:

5 000 ppm

System:

other: Apart from decreased body weight gain - none

Organ:

other: no obvious target identified (screening level study)

Treatment related:

yes

Dose response relationship:

yes

Overall reproductive toxicity

Reproductive effects observed:

yes

Lowest effective dose / conc.:

5 000 ppm

Treatment related:

yes

Relation to other toxic effects:

not specified

Dose response relationship:

yes

Any other information on results incl. tables

Table 1: Estrous cycle pre-mating

		Group 0		Group 1	Group 2	Group 3
		0 ppm		500 ppm	1500 ppm	5000 ppm
Number of Cycles	Mean	4.2	k	4.3	4.5	3.9
	S.d.	0.4		0.5	0.5	0.9
	N	10		10	10	10
Cycles Length (days)	Mean	4.0	v	4.0	4.0	4.3
	S.d.	0.0		0.0	0.1	0.5
	N	10		10	10	10
Cycling Normally (3-6 Days)	N	10		10	10	10
	%	100.0		100.0	100.0	100.0
Long Estrous (3 Days)	N	0		0	0	0
	%	0.0		0.0	0.0	0.0
Long Diestrous (4 Days)	N	0		0	0	1
	%	0.0		0.0	0.0	10.0

Statistic Profile = Kruskal-Wallis + Wilcoxon test (two-sided), * p<=0.05, ** p <=0.01, X = Group excluded from statistics

k=KRUSKALL-WALLIS; v=KRUSKALL-WALLIS-WILCOX

Table 2: Summary mating report

		Group 0		Group 1	Group 2	Group 3
		0 ppm		500 ppm	1500 ppm	5000 ppm
No. of females mated	N	10		10	10	10
- Inseminated	N	10	f-	10	10	10
Female mating index	%	100.0		100.0	100.0	100.0
-- Pregnant	N	9	f-	10	9	10
Female fertility index	%	90.0		100.0	90.0	100.0
No. of males mated	N	10		10	10	10
- With inseminated females	N	10	f-	10	10	10
Male mating index	%	100.0		100.0	100.0	100.0
- With pregnant females	N	9	f-	10	9	10
Male fertility index	%	90.0		100.0	90.0	100.0
Females with defined Day 0 pc	N	10		10	10	10
Mating days until Day 0 pc	Mean	2.4	x+	4.0	2.4	2.6
	S.d.	1.1		3.2	1.2	1.4
	N	10		10	10	10
Days 0 To 4	N	10		9	10	9
	%	100.0		90.0	100.0	90.0
Days 5 To 9	N	0		0	0	1
	%	0.0		0.0	0.0	10.0
Days 10 To 14	N	0		1	0	0
	%	0.0		10.0	0.0	0.0

Statistic Profile = Fisher's exact test (one-sided-), Wilcoxon with Bonferroni-Holm (one-sided+), * p<=0.05, ** p <=0.01, X = Group excluded from statistics

f=FISHER-EXACT; x=WILCOX

Table 3: Summary of pregnancy status report

		Group 0	Group 1	Group 2	Group 3
		0 ppm	500 ppm	1500 ppm	5000 ppm
No. of females at start	N	10	10	10	10
No. of females mated	N	10	10	10	10
Without evidence of mating	N	0	0	0	0
Females with defined Day 0 pc	N	10	10	10	10
Pregnant	N	9	10	9	10
- sacrificed scheduled	N	9	10	9	10
Not pregnant	N	1	0	1	0
- sacrificed scheduled	N	1	0	1	0
Pregnant, not delivering	N	0	0	0	0
Delivering	N	9	10	9	10
-- With liveborn pups	N	9	10	9	10
	%	100.0	100.0	100.0	100.0
-- With all pups stillborn	N	0	0	0	0
	%	0.0	0.0	0.0	0.0

Table 4 Summary delivery report

		Group 0	Group 1	Group 2	Group 3
		0 ppm	500 ppm	1500 ppm	5000 ppm
Gestation Index	%	100.0	100.0	100.0	100.0
Gestation days	Mean	22.3 n	21.9	21.7**	22.1
	S.d.	0.5	0.3	0.5	0.3
	N	9	10	9	10
-- With stillborn pups	N	0 f+	2	0	2
	%	0.0	20.0	0.0	20.0
-- With all pups stillborn	N	0 f+	0	0	0
	%	0.0	0.0	0.0	0.0

Statistic Profile = Fisher's exact test (one-sided-), Dunnett test (two-sided), Fisher's exact test (one-sided+), * p<=0.05, ** p <=0.01, X = Group excluded from statistics

f=FISHER-EXACT; n=DUNNETT

Table 5: Summary Litter report - pup status

			Group 0		Group 1	Group 2	Group 3
			0 ppm		500 ppm	1500 ppm	5000 ppm
Total Number of Pregnant Females	N		9		10	9	10
Total number of litters	N		9		10	9	10
With liveborn pups	N		9	f-	10	9	10
	%		100.0		100.0	100.0	100.0
With stillborn pups	N		0	f+	2	0	2
	%		0.0		20.0	0.0	20.0
With all pups stillborn	N		0	f+	0	0	0
	%		0.0		0.0	0.0	0.0
Implantation Sites	N		130		131	98	99
Historical Control Range 9.8-14.2	Mean		14.4	x-	13.1	10.9**	9.9**
	S.d.		1.9		2.1	1.6	2.3
	N		9		10	9	10
Pups delivered	N		113		129	92	87
Historical Control Range 9.0 - 13.2	Mean		12.6	x-	12.9	10.2*	8.7**
	S.d.		2.2		2.1	1.7	2.1
	N		9		10	9	10
Postimplantation Loss	Mean%		12.1	x+	1.6	5.7	10.4
	S.d.		15.7		3.5	11.1	16.1
	N		9		10	9	10

Statistic Profile = Wilcoxon with Bonferroni-Holm (one-sided-), Wilcoxon with Bonferroni-Holm (one-sided+), Wilcoxon test (two-sided), Fisher's exact test (one-sided-),

Fisher's exact test (one-sided+), * p<=0.05, ** p <=0.01, X = Group excluded from statistics

f=FISHER-EXACT; x=WILCOX

Table 6:  Thyroid hormones, F0 males

		Group 0	Group 1	Group 2	Group 3
		0 ppm	500 ppm	1500 ppm	5000 ppm
T4	Mean	62.84 k	63.11	63.39	57.75
[nmol/L]	S.d.	9.44	6.72	10.77	8.75
day 94	N	10	10	10	10
	Median	67.60	62.18	63.14	60.31
	Deviation Vs Control [%]		0.43	0.88	-8.09
TSH	Mean	5.94 k	6.51	8.58	8.64
[µg/L]	S.d.	1.74	2.57	3.23	2.94
day 94	N	10	10	10	10
	Median	6.12	6.01	7.93	8.78
	Deviation Vs Control [%]		9.50	44.41	45.37

Table 7: Absolute and relative organ weights

Relative weights	Males			Females
Test group (ppm)	1 (500)	2 (1500)	3 (5000)	1 (500)	2 (1500)	3 (5000)
Brain	102%	107%	116%**	99%	101%	114%*
Kidneys	116%**	110%*	104%	102%	93%	114%**
Liver	103%	112%**	127%**	105%*	112%**	125%**
Ovaries				93%	85%	69%**
Thyroid glands	101%	100%	119%*
Absolute weights	Males			Females
Test group (ppm)	1 (500)	2 (1500)	3 (5000)	1 (500)	2 (1500)	3 (5000)
Final body weight	97%	96%	87%**	102%	97%	87%**
Epididymides	98%	94%	91%*
Heart	97%	93%	87%*	103%	93%*	87%*
Kidneys	111%*	101%	87%*	103%	89%*	97%
Ovaries				94%	82%*	60%**

* : p <= 0.05, **: p <= 0.01