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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
three-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
no data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Well documented study with the following deviations: Only one dose level was included in the study. Test material was not analyzed in diet. No impurity profile of test material was provided. Source of animals was not specified. The group size was smaller than the 20 females and 10 males required in OECD guideline 416. There was no recorded acclimation period for the P generation. Only one dose tested. Daily observation of clinical signs of parents and observations of abnormal behaviour of offspring are not reported. Body Weights: parental body weights were not recorded during gestation and lactation. Food consumption was not determined. Estrus cycle was not evaluated. Sperm parameters were not included. Offspring sex and sex ratios were not determined. Litter body weight was reported; however, individual pup body weights were not recorded. There is no report of gross pathology and necropsy observations. Organ weight data for the uterus, ovaries, epididymides, prostate, seminal vesicles, pituitary gland, adrenals and thyroid gland were not included. Histopathology was conducted only on the F3b offspring and did not include the following organs: uterus, cervix, vagina, epididymides, seminal vesicles, prostate, coagulating gland, pituitary gland. Note that the known target organ (kidney) was evaluated. Statistics are not reported. Study has been reviewed by an expert assessor (see expert report attached) and determined to be suitable for use for the purposes of REACH registration.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1960
Report Date:
1960

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Groups of male (8/group) and female (16/group) rats were fed control diet or diet treated with 0.05% test material for 100 days (P1) generation. Body weights were measured weekly during the 100 day pre-mating exposure for each generation. Each male was caged with two females for 7 days until conception occurred. Pregnancy rates were determined. The first litter born of the parental animals was designated as the F1a litter. Upon delivery, offspring were counted and average litter weights determined. Pup survival was determined on days 1-5 and days 6-21. On day 5, pups were culled to 10 per litter. Body weights per litter were determined on day 21. The F1a litter was sacrificed at 30 days postnatally. Parental animals were mated again 10 days after the sacrifice of the F1a litter (ten-day interim period). Mating took place for 7 days. A second litter (F1b) was born after approximately 21 days of gestation. Upon delivery, offspring were counted and the average litter weights determined. Pup survival was determined on days 1-5 and days 6-21. On day 5 pups were culled to 10 per litter. Body weights per litter were determined on day 21. Following weaning of the F1b litter, 8 males and 16 females from the F1b litter from the control group and the treated group were fed their respective diets for 100 days. Again two litters (F2a and F2b) were produced in the same sequence described for the F1 litters. Parental animals were mated for 7 days after the ten-day interim period after the sacrifice of the F2a litter (pups were 30 days old). Following birth of the F2b litter the same offspring and parental parameters were measured. Eight males and 16 females of the F2b litter were designated for continual exposure for another 100 days. Upon completion of the exposure of the F2b litter, animals were mated again to produce the F3a and F3b litters in the same manner described above. The ten males and ten females from the final litter, the F3b litter, were evaluated for body weight, histopathology (liver, kidneys, gonads, lungs, brain, stomach, heart, spleen, adrenal, stomach, large and small intestines, bladder, bone marrow, muscle, pancreas, lymph nodes, gut) and organ weights (liver, kidneys, testes, lungs, brain, stomach, heart, spleen). Individual animal data were collected and recorded throughout the study.
GLP compliance:
no
Remarks:
study predates GLP
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: granules
Details on test material:
- Name of test material (as cited in study report): Sodium Trimetaphosphate
- Substance type: Granular
- Physical state: Solid
- Analytical purity: no data
- Lot/batch No.: Lot #195001-8, E9309 and 4-CEC
- Stability under test conditions: Test material is stable at room temperature
- Other: Specification: Granular sodium trimetaphosphate from Calgon, Inc.

Test animals

Species:
rat
Strain:
other: Rochester Strain (Ex-Wistar 1923)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: (P) x wks; (F1) x wks
- Weight at study initiation: Body weight was approximately 60 grams for males and 59 grams for females at the start of the 100 day exposure.
- Fasting period before study: no data
- Housing: no data
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: not applicable


Administration / exposure

Route of administration:
oral: feed
Type of inhalation exposure (if applicable):
other: not applicable
Vehicle:
unchanged (no vehicle)
Details on exposure:
No data
Details on mating procedure:
- Length of cohabitation: 7 days
- Any other deviations from standard protocol: Litters were culled to ten pups on day 5 of lactation.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
no data
Duration of treatment / exposure:
Exposure took place for 100 days prior to mating, 21 days of gestation and during lactation. Litters were culled to ten pups with approximately 5/sex/group on postnatal day 5. For the F1, F2, F3-generations, offspring were weaned and exposed during an interim period of 10 days and then during a mating period of 7 days. Dams were exposed continually during gestation and lactation. Offspring of the second litter were again mated for 7 days after a 10 day interim period following weaning. The first litters (F1a, F2a, F3a) were sacrificed at 30 days of age. The second litters (F1b, F2b) were used to produce the succeeding generation. The F3b pups were used for the histopathology evaluation and organ weight determinations.
Frequency of treatment:
daily, ad libitum
Details on study schedule:
Beginning with the original groups of rats, 16 female and 8 male rats were removed at the age of 100 days and mated. The matings were carried out in such a way that male rats were rotated through cages each housing two female rats.

For the F1, F2, F3-generations, offspring were weaned and exposed during an interim period of 10 days and then during a mating period of 7 days.
Offspring of the second litter were again mated for 7 days after a 10 day interim period following weaning.

The first litters (F1a, F2a, F3a) were sacrificed at 30 days of age. The second litters (F1b, F2b) were used to produce the succeeding generation. The F3b pups were used for the histopathology evaluation and organ weight determinations.

Litters were culled to ten pups with approximately 5/sex/group on postnatal day 5.
Doses / concentrations
Dose / conc.:
0.05 other: % w/w nominal in diet
No. of animals per sex per dose:
P1, F1, F2, F3: 16 females and 8 males
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale:
The reproduction study began with 16 female and 8 male rats from the control diet group and from the group given the diet containing 1% of TMP in the 2-year study. When it was discovered that the male rats given a 1% Trimetaphosphate diet were not maintaining normal growth, it was obvious that this level was unsuitable one for reproduction study. Consequently additinoal diet groups were started at 0.1% and at 0.05% to be sure that no detectable effect could be laid to the TMP in the diet. The continued stress of repeated production of generations and litters of rats could then be tested. In 1957 it was decided to conduct a reproduction study on rats maintained on the diet containing 0.05% trimetaphosphate.

- Rationale for animal assignment (if not random): no data

- Other: no data
Positive control:
no data

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: no data
- The animals were monitored for general well-being.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly during the 100 day pre-mating period, but not evaluated during gestation and lactation.
Oestrous cyclicity (parental animals):
Not evaluated.
Sperm parameters (parental animals):
The following were not evaluated: testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, enumeration of cauda epididymal sperm reserve, sperm motility, sperm morphology.
Litter observations:
number of pups, stillbirths, live births, weight gain per litter

The following was not evaluated: sex of pups, presence of gross anomalies, physical or behavioural abnormalities. No individual pup body weights.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals.
- Maternal animals: All surviving animals.

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
ORGAN WEIGHTS; F3b:
liver, kidneys, testes, lungs, brain, stomach, heart, spleen on 10/sex/group from the F3b offspring at 21 days of age.
The following was not evaluated: Uterus, ovaries, epididymides (total and cauda), prostate, seminal vesicles, pituitary, thyroid, adrenal glands

HISTOPATHOLOGY; F3b:
Kidneys, heart, brain, spleen, lungs, liver, ovaries, testes, adrenal, urinary bladder, stomach, large and small intestines, muscle, bone marrow, muscle, pancreas, gut and lymph noted on F3b animals.
The following was not evaluated: Vagina, uterus, epididymis, seminal vesicle, prostate
coagulating gland.
Not evaluated

Results and discussion

Results: P0 (first parental generation)

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
0.05 other: % w/w
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted

Target system / organ toxicity (P0)

Key result
Critical effects observed:
no

Results: F1 generation

Effect levels (F1)

open allclose all
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
0.05 other: % w/w
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects notes
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted
Remarks on result:
other: calculated

Target system / organ toxicity (F1)

Key result
Critical effects observed:
no

Results: F2 generation

Effect levels (F2)

open allclose all
Key result
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
0.05 other: % w/w
Based on:
test mat.
Sex:
male/female
Remarks on result:
other: no effects noted
Key result
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
ca. 25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted
Remarks on result:
other: calculated

Target system / organ toxicity (F2)

Key result
Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Any other information on results incl. tables

NOAEL calculated in accorance with Appendix F, guidelines for the preparation of toxicological working papers for the joint FAO/WHO expert committee on food additives, December 2000.

This study included only one dose level of the test material, 0.05% in the diet. Table 1 summarizes results on many parameters. There was no treatment-related effect on litter weights. The fertility index, duration of pregnancy, live birth index, litter size, litter weights, average pup weights, number of pups per litter, survival index, viability index and lactation index were all comparable between control and treated groups. Organ weights were similar between control and treated groups. Treated females of the F3b generation had somewhat lower spleen weights than control animals. There were no treatment-relatedhistopathologicalfindings.

 

Table 1. Reproductive toxicity

Parameter

 

Control

High dose

Generation

M

F

M

F

Mortality

Incidence

P

0

0

0

0

 

 

F1

3

1

0

0

 

 

F2

0

2

0

0

 

 

F3

0

1

0

1

Food consumption

% of control

 

NA

NA

NA

NA

Body weight gain

% of control

 

100

100

100

100

Clinical Observations

specify effects

Incidence

 

NA

NA

NA

NA

Organ weights (F3b)

Liver

Kidneys

Testes

Lungs

Brain

Stomach

Heart

Spleen

% of control

 

 

 

 

 

 

 

 

95.6

98.9

92.1

95.4

100.6

88.0

89.4

83.7

 

96.8

91.9

-

93.4

105.5

92.5

90.5

69.2

Pathology

 

 

 

 

 

 

Histopathologicexamination: F3b

Incidence

0

0

0

0

0

 

Applicant's summary and conclusion

Conclusions:
The test material did not cause any adverse effects on fertility, reproductive performance, offspring viability, offspring survival and offspring body weight in male and female rats when administered continuously for three generations in the diet at a dose level of 0.05% in the diet. The test material did not cause any treatment-related effects on organ weights or any histopathological findings on the tissues evaluated at a dose level of 0.05% in the diet.