A mixture of: α-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyl-ω-hydroxypoly(oxyethylene); α-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyl-ω-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyloxypoly(oxyethylene)

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May. 7, 1990 to Mar. 22, 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study following OECD guideline 415 without deviations.

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Reference 2

Reference Type:

other: Peer review on pathological data

Title:

Unnamed

Year:

1991

Report date:

1991

Reference 3

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Version / remarks:

(1983)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

A mixture of: α-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyl-ω-hydroxypoly(oxyethylene); α-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyl-ω-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyloxypoly(oxyethylene)

EC Number:

400-830-7

EC Name:

A mixture of: α-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyl-ω-hydroxypoly(oxyethylene); α-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyl-ω-3-(3-(2H-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl)propionyloxypoly(oxyethylene)

Cas Number:

104810-48-2

IUPAC Name:

14-({3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoyl}oxy)-3,6,9,12-tetraoxatetradecan-1-yl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 14-hydroxy-3,6,9,12-tetraoxatetradecan-1-yl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; tris(17-({3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoyl}oxy)-3,6,9,12,15-pentaoxaheptadecan-1-yl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate); tris(17-hydroxy-3,6,9,12,15-pentaoxaheptadecan-1-yl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate); 2-(2-hydroxyethoxy)ethan-1-ol; 2-({3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoyl}oxy)ethyl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 2-[2-(2-hydroxyethoxy)ethoxy]ethan-1-ol; 2-[2-(2-hydroxyethoxy)ethoxy]ethyl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 2-hydroxyethyl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 2-methoxyethan-1-ol; 2-{2-[2-({3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoyl}oxy)ethoxy]ethoxy}ethyl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 20-({3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoyl}oxy)-3,6,9,12,15,18-hexaoxaicosan-1-yl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 20-hydroxy-3,6,9,12,15,18-hexaoxaicosan-1-yl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate; 3,6,9,12,15,18,21-heptaoxatricosane-1,23-diol; 3,6,9,12,15-pentaoxaheptadecane-1,17-diol; 3,6,9,12-tetraoxatetradecane-1,14-diol; bis(ethane-1,2-diol)

Details on test material:

- Physical appearance: liquid (yellow, high viscous)
- Lot/batch No.: EN.20043.42
- Expiration date of the lot/batch: 12/1994
- Purity of test article: was confirmed (see Analytical Certificate No. 532, method KB-96/1 Appendix 26 and 27)
- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Strain as stated in the report: Tif: RAif (SPF), hybrids of RII/1 x RII/2
- Source: Animal Production, WST-455 CIBA-GEIGY Limited, 4332 Stein Switzerland
- Age at study initiation: ca. 6-7 weeks
- Weight at study initiation: males: ca. 174 g; females: ca. 144 g
- Housing: individually in Macrolon cages
- Diet ad libitum: pelleted, certified standard diet (Nafag No. 890 Tox, Nafag, Naehr- und Futtermittel AG, Gossau, Switzerland)
- Water ad libitum: tap water
- Acclimation period: at least one week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 50 ± 15
- Air changes (per hr): 16
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on exposure:

PREPARATION OF DOSING SOLUTIONS
Test item was mixed with the vehicle and administered orally (by gavage). During dosing the homogeneity of the test article mixtures was maintained by means of a magnetic stirrer. The mixtures contained 0, 0.4, 10, and 20 mg test article/ml vehicle for the control (Group 1), low (Group 2), intermediate (Group 3) and high dose groups (Group 4), respectively.

Details on mating procedure:

One male and one female were caged together overnight. Successful mating was recorded by the presence of a vaginal plug or the presence of sperm in vaginal smear. The day of successful mating was designated day "0" of pregnancy (or day "0" post coitum).
In the case of unsuccessful mating, the mating procedure was repeated daily for a maximum of 21 days. In case a male died prior to mating, another male from the same dose group that has already successfully mated replaced the male that died.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Every day prior to administration, one sample was taken from the vehicle control and three samples (one each from the top, middle and bottom of the container) were taken from each dose concentration. In addition, one sample was taken from each dose concentration at the end of the dosing procedure. The purpose of the analytic analysis was to verify the stability of test article/vehicle mixture, the homogeneity of the test article in the vehicle and achieved concentrations of the prepared dosing mixtures. Results of analytical investigations demonstrated that the preparation of the dosing mixtures was accurate and that the active ingredient was stable in the administration form during the dosing period.

Duration of treatment / exposure:

For males 16 weeks
For females 11 weeks

Frequency of treatment:

daily (7 days a week)

Details on study schedule:

Males and females received the test article prior to mating (70 and 14 days, respectively) and throughout mating. Males were further treated until parturition of dams and females were treated until parturition and during gestation and lactation (day 21 post partum).

Doses / concentrations

Remarks:

Doses / Concentrations:
2, 50, 100 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

25

Control animals:

yes, concurrent vehicle

Details on study design:

The dose selection based on a range finding toxicity study with doses of 0, 20, 100, and 200 mg/kg bw/d in 8 males and 8 females (Ciba-Geigy 894553).

Examinations

Parental animals: Observations and examinations:

CLINICAL SIGNS AND MORTALITY
The animals were observed daily for clinical signs and mortality.
BODY WEIGHT
The body weight was recorded daily during premating period and gestation/lactation period. No weighing was performed during mating period.
FOOD CONSUMPTION
The food consumption was recorded weekly during the premating period and the gestation/lactation period but not during mating period.
REPRODUCTION PARAMETERS
The following maternal reproduction parameters were reported:
- number of pregnant animals (with and without pups at delivery)
- number of implantations
- prenatal loss: number of implantations minus number of live and dead young on day 0 p.p. as a percentage of the implantations
- number of dams with only live or still births and number of dams with labor disturbances
- number of dams with live pups at delivery and the length of gestation period for these dams
- total litter size on day 0 p.p.
- litter weight on day 0 p.p. (by weighing individual body weights)

Litter observations:

The following parameters of F1 offspring were observed:
- live birth index (%)
- perinatal loss (%)
- postnatal loss of young between days 0 to 4, 4 to 7, 7 to 14 and 15 to 21 p.p. as well as the overall loss between days 1 to 21 p.p.
- viability index on day 4 p.p.
- sex (on the basis of ano-genital distance)
- individual body weight of live young on days 4, 7, 14 and 21 p.p.
- lactation index (%)
During the pre-weaning period all offspring in all litters were examined to determine the age at which the following physical developmental stages were attained:
- eye opening: the day on which the upper and lower eyelids of both eyes first separate
- beginning of hair growth: the day on which growth of body hair is first observed macroscopically
- eruption of incisors: the day on which the upper incisor is first observed to penetrate through the gum
The following reflex tests were included:
- surface righting reflex
- startle reflex
- pupil reflex

Postmortem examinations (parental animals):

SACRIFICE
At the end of the treatment period male animals were sacrificed and subjected to gross necropsy. Dams and their litters were sacrificed and subjected to necropsy at the end of lactation period (day 21 post partum). All animals were exsanguinated under ether anesthesia.
NECROPSY
Gross pathology of all males and females included detailed pathological examination of the organs listed in Table 1. Samples were preserved in neutral buffered 4% formalin for histopathological examinations.
Organ weights of liver, kidneys, testis and ovaries were recorded.
The following organs and tissues of parental animals of all treated groups were examined histopathologically: Liver, kidneys, vagina, uterus, both ovaries and mammary area of females, as well as liver, kidneys, both testis, both epididymis, seminal vesicle and prostate of male rats.
ELECTRON MICROSCOPY AND BIOCHEMICAL INVESTIGATIONS
Samples of liver from 40 selected parental males were collected, fixed and embedded for electron microscopy. Further liver samples from these animals were frozen in liquid nitrogen for further specific biochemical analysis. The results of electron microscopy and biochemical analyses were reported in a separate study report (Ciba-Geigy CB 90/23 BY). For further information see IUCLID chapter 7.9.3 specific investigations. It was mentioned in the report that, due to spontaneous death of some males which were intended for liver tissue sampling (electron microscopy and biochemical investigations), these animals had to be replaced with another animals from the same groups.

Postmortem examinations (offspring):

Dead pups were examined for gross external anomalies.

Statistics:

Statistical analysis of continuous data was performed using the Analysis of Variance Procedure (ANOVA) followed by Dunnett's t-Test in case of a significant result in the ANOVA. Categorical data (e.g. malformation counts) were analysed using Chi-Square test followed by Fisher's Exact test in case of a significant result in the Chi-Square test. Mean percent data were analysed using the Kruskal-Wallis nonparametric analysis of variance test followed by Mann-Whitney U-test.
However, a statistically significant difference between two values does not necessarily imply biological relevance of that deviation.

Offspring viability indices:

- viability index on day 4 p.p. (%) = number of viable offspring on day 4 p.p. x 100 / number of viable offspring on day 0 p.p.
- live birth index (%) = number of viable offspring on day 0 p.p. x 100 / total number of offspring
- perinatal loss (%) = number of dead offspring on day 0 p.p. x 100 / total number of offspring
- prenatal loss (%) = number of implantations minus number of live and dead young on day 0 p.p. as a percentage of the implantations.

Results and discussion

Results: P0 (first parental generation)

Details on results (P0)

CLINICAL OBSERVATIONS AND MORTALITY (MALES)
Clinical symptoms as dyspnea were observed sporadically in animals of all groups and were not considered to be related to the test article. Mortalities were not treatment related. For details see Table 1.
BODY WEIGHT (MALES)
At 100 mg/kg bw/day treatment related decreases in absolute male body weight were observed from day 69 (premating) until day 119 (see Table 2).
At 50 mg/kg bw/d treatment related decreases in absolute male body weight were observed from day 76 (mating) until day 119(see Table 2).
At 2 mg/kg bw/d no treatment related decreases in absolute male body weight were observed.
Treatment related reductions in body-weight gain were observed at dose levels of 50 (27%) and 100 mg/kg bw/d (36%).
The differences were statistically significant at 100 mg/kg bw/d for the intervals day 29 to 36 (16%), day 43 to 50 (7.1%), day 50 to 57 (8.4%), day 57 to 64 (7.2%), and day 64 to 70 (-1.9%), day 70 to 78 (-2.5%), day 85 to 92 (6.6%), day 92 to 99 (1.6%)
At 50 mg/kg bw/d differences were statistically significant at days 29 to 36 (17%), 43 to 50 (11.1%), 64 to 70 (2.1%) and 92 to 99 (2.6%).
FOOD CONSUMPTION (MALES)
Mean daily food consumption for male animals at 100 mg/kg bw/d was significantly increased at different time intervals over the whole treatment period of 16 weeks. As example for weeks 3 to 4 mean food consumption for the treated males was 28 g/animal/d versus 26 g/animal/d at control. The reason for this increase is not known. In the lower dose groups, food consumption was comparable to control values throughout the study.
GROSS PATHOLOGY (MALES)
At 50 and 100 mg/kg bw/d enlarged liver was observed at necropsy (7/24 and 17/24 males, respectively.) This finding was considered treatment related. Other findings like mottled lung were seen sporadically and were not treatment related.
ORGAN WEIGHTS (MALES)
At 50 and 100 mg/kg bw/d the liver weight (25 g and 29 g compared to 17 g at control) as well as the liver to body weight ratio (0.06 and 0.07 compared to 0.04 at control) was increased. The kidney weight (3.1 g and 3.2 g compared to 2.9 g at control) and kidney to body weight ratio (0.007 and 0.007 compared to 0.006 at control) was also increased in both treatment groups.
MICROSCOPICAL FINDINGS (MALES)
At 50 and 100 mg/kg bw/d hepatocyte hypertrophy was found (23 males of each group). Additionally, necrosis of hepatocytes or multifocal organizing necrosis of the liver parenchyma, was reported in all males of both groups. At 50 and 100 mg/kg bw/d the majority of animals showed accumulation of iron-positive pigment within the cytoplasm of some kupffer cells, macrophages and hepatocytes.
At 50 and 100 mg/kg bw/d hepatocyte hypertrophy was found (7 and 13 females, respectively).
A peer review by Hess (Report no. 894538, 1991) stated that focal hepatitis occurred in animals treated with 50 or 100 mg/kg bw/d, however in a much lesser extend in the higher dosed animals. According to the author this renders the experimental significance of the organizing and recent necrosis doubtful. However, the incidence of liver necrosis and hepatocyte hypertrophy in males was generally confirmed by the review.
REPRODUCTIVE PERFORMANCE (MALES)
Neither insemination nor fertility was affected by treatment with the test article. There was no effect of treatment on the time required (duration) until mating occurred. Neither macroscopical nor histopathological examination revealed any treatment related changes in the male reproductive system at any dose level.
CLINICAL OBSERVATIONS AND MORTALITY (FEMALES)
Clinical symptoms such as crusts/shorfs or diarrhea were observed sporadically in animals of all groups and were not considered to be related to the test article. Mortalities were not treatment related. For details on mortality see Table 1.
BODY WEIGHT (FEMALES)
At 50 mg/kg bw/d the mean maternal body weight was increased at day 12 of the premating phase and on day 0 post coitum (190 and 206 g compared to 181 and 196 g in control animals).
At 100 mg/kg bw/d the mean maternal body weight was increased at day 10 and 12 of the premating phase and on day 0 post coitum (181, 188 and 204 g compared to 174, 181 and 196 g in control animals).
Treatment related increased body-weight gain was observed at dose levels of 50 mg/kg bw/d (33%) and 100 mg/kg bw/d (27%) during premating for days 1 to 8. At 100 mg/kg bw/d the body-weight gain was decreased for days 0 to 7 p.c. (16%) during gestation and days 0 to 7 during lactation (34%).
FOOD CONSUMPTION (FEMALES)
At 50 mg/kg bw/d the mean food consumption was significantly increased during gestation between days 14 to 21 p.c. (27 g/animal/d compared to 25 g/animal/d).
At 100 mg/kg bw/d the mean food consumption was significantly increased during gestation between days 14 to 21 p.c. (26 g/animal/d compared to 25 g/animal/d) and decreased during lactation between days 7 to 14 and days 14 to 21 (58 g/animal/d compared to 66 g/animal/d and 71 g/animal/d compared to 87 g/animal/d).
GROSS PATHOLOGY (FEMALES)
No treatment related pathological changes were recorded in females. Findings like dilation of caecum were observed in females of all groups or sporadically (e.g. hair loss) and were therefore not treatment related.
ORGAN WEIGHTS (FEMALES)
At 50 and 100 mg/kg bw/d the liver weight (21 g and 21 g compared to 17 g at control) as well as the liver to body weight ratio (0.07 and 0.07 compared to 0.06 at control) was significantly increased. Additionally, at 100 mg/kg bw/d the kidney weight (2.4 g compared to 2.3 g at control) was significantly increased.
MICROSCOPICAL FINDINGS (FEMALES)
At 50 and 100 mg/kg bw/d hepatocyte hypertrophy was found (7/24 and 13/24 females, respectively). Additionally, necrosis of hepatocytes or multifocal organizing necrosis of the liver parenchyma, was reported in 3/24 and 2/24 females (50 and 100 mg/kg bw/d).
As for males the liver findings had been peer reviewed (Hess, 1991). The author of the peer review did not confirm the occurence of hepatocyte necrosis in 3 females. Therefore, hepatocyte single cell necrosis was confined to 1 animal of the 50 and to 1 animal of the 100 mg/kg bw/d group. The reviewer stated that the experimental significance of this finding is doubtful.
REPRODUCTIVE PERFORMANCE AND PREGNANCY OUTCOME
Neither insemination nor fertility was affected by treatment with the test article. There was no effect of treatment on the time required (duration) until mating occurred. No treatment related effect on the percentage of females that maintained pregnancy, on the mean duration of gestation, on abortions or early deliveries were seen. The number of females with stillborn pups was increased at 2, 50 and 100 mg/kg bw/d (2/22, 5/22 and 9/23 females compared to 0/24 at control).
Neither macroscopical nor histopathological examination revealed any treatment related changes in the female reproductive system at any dose level.

Effect levels (P0)

1

Dose descriptor:

NOAEL

Remarks:

(systemic toxicity)

Effect level:

2 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

2

Dose descriptor:

LOAEL

Remarks:

(systemic toxicity)

Effect level:

50 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The LOAEL is based on effects on body weight, food consumption and liver (weight and histopathological changes) in male and females, on effects on kidney (weight and/or histopathological changes) in males.

3

Dose descriptor:

NOAEL

Remarks:

(fertility)

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Results: F1 generation

Details on results (F1)

VIABILITY (OFFSPRING)
At 50 mg/kg bw/d the number of stillborn pups was significantly increased (perinatal loss = 3% compared to 0% at control) which consequently led to a decreased live birth index (97% compared to 100% at control).
At 100 mg/kg bw/d prenatal loss was increased resulting in a decreased mean litter size (12.6 pups/litter compared to 13.9 at control). The number of stillborn pups was significantly increased (perinatal loss = 7% compared to 0% at control) which consequently led to a decreased live birth index (93% compared to 100% at control). In addition, the pup mortality was significantly increased postnatal during days 1 to 4 (15/290 pups died compared to 3/334 at control). The increased mortality of pups at 100 mg/kg bw/d lead to a reduced number of live pups per litter (11.7 compared to 13.9 in the control group on day 0 p.p. and 10.8 compared to 13.7 in the control group on day 21 p.p.).
BODY WEIGHT (OFFSPRING)
At 100 mg/kg bw/d the birth weight of male and female pups, and the mean pup weight during the lactation period was decreased compared to the control (see table 3).
SEXUAL MATURATION (OFFSPRING)
Pup sex ratios were comparable between groups. There was no evidence of selective mortality in male or female pups.
GROSS PATHOLOGY (OFFSPRING)
At 50 mg/kg bw/d, 2 of 300 pups had dark skin of abdomen and hind limbs or tail at birth. The dark skin disappeared in 1 case between days 2 and 8 p.p. The other pub had a necrotic tail on day 21 p.p.
At 100 mg/kg bw/d 26 of 290 pups had dark skin of abdomen, right and/or both hind limbs and/or tail at birth. In 11 cases the dark skin disappeared between days 2 and 8 p.p.; these pups survived to weaning. In 4 cases a necrotic tail on day 21 p.p. was seen. The remaining 11 pups died between days 1 and 2 p.p. A delay of physical development (tooth eruption, eye opening) was seen at 100 mg/kg bw/d.

Effect levels (F1)

1

Dose descriptor:

NOAEL

Remarks:

(offspring)

Generation:

F1

Effect level:

2 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

2

Dose descriptor:

LOAEL

Remarks:

(offspring)

Generation:

F1

Effect level:

50 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The LOAEL is based on a reduced live birth index (97% and 93% at the mid and high dose group versus 100% in control) and dark abdominal skin of pups in 10% of the high dose dams.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Table 1: Summary of mortality data.

MALES
Group mg/kg bw/d	No. of dead animals	Remarks	Treatment related
0	1	Intubation error	No
2	3	Intubation error, moribund, incidental	No
50	1	Intubation error	No
100	2	Intubation error, incidental	No
FEMALES
Group mg/kg bw/d	No. of dead animals	Remarks	Treatment related
0	2	Intubation error	No
2	2	Intubation error, moribund (premating),	No
50	1	Intubation error	No
100	1	Intubation error	No

 

Table 2: Treatment related changes in body weight (g)

MALES
day	Dose (mg/kg bw/d)
	0	2	50	100
69	417	409	399	390*
70	416	409	398	390*
71	415	407	395	385*
72	415	407	396	385*
73	418	408	398	388*
74	419	409	399	386**
75	420	410	400	385**
76	421	411	400*	385**
77	424	414	400*	386**
78	423	412	402*	388**
79	423	413	400*	387**
80	426	414	402*	389**
81	426	414	401*	389**
82	427	416	402*	390**
83	429	418	404*	393**
84	431	419	405*	395**
85	431	420	404*	394**
86	433	420	406*	394**
87	433	422	406*	395**
88	436	425	408**	395**
89	437	425	408**	395**
90	440	427	411**	399**
91	441	430	413*	398**
92	442	430	413**	401**
93	443	432	416*	401**
94	442	432	417*	402**
95	445	436	417*	401**
96	449	438	418**	402**
97	449	436	416**	404**
98	449	439	416**	403**
99	449	439	416**	402**
100	451	438	416**	403**
101	453	439	416**	403**
102	453	440	416**	403**
103	453	440	417**	406**
104	457	442	420**	410**
105	457	444	427**	415**
106	458	444	424**	415**
107	459	445	427**	418**
108	459	446	428**	420**
109	458	446	429*	420**
110	460	446	431*	420**
111	463	451	432**	422**
112	464	451	436*	423**
113	466	452	435**	420**
114	464	451	437*	427**
115	466	453	438*	427**
116	465	452	438*	426**
117	466	452	439*	425**
118	467	453	438*	426**
119	469	454	438*	426**
*, p>0.05; **, p>0.01

Table 3: Mean pup weight (g)

MALE AND FEMALE PUPS
day	Dose (mg/kg bw/d)
	0	2	50	100
0	6.2	6.2	6.0	5.3**
4	9.7	9.8	9.6	8.7**
7	13.9	14.2	13.8	12.3**
14	26.0	25.3	25.7	23.7**
21	44.8	43.4	43.4	39.7**