Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

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

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1960
Report Date:
1960
Reference Type:
publication
Title:
Summaries of toxicological data
Author:
Summaries published by BIBRA
Year:
1964
Bibliographic source:
Fd Cosmet. Toxicol. Vol. 2 pp. 147-154. Pergamon Press 1964.

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
A reproduction study spanning 3 generations of rats reared and maintained on diets containing 0.0% and 0.5% HMP (Hexametaphosphate). Per generation two different litters were produced; one was sacrificed and the other was bred to produce the next generation. Matings were carried out between 16 females and 8 males in each group when the rats were 100 days old. Observations were limited to: the numbers of pups born, pup mortality up to 21 days, organ weights and histological changes of tissues and organs.
GLP compliance:
no
Remarks:
study predates GLP
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Sodium hexametaphosphate
- Substance type: granular
- Physical state: solid
- Analytical purity: no data
- Lot/batch No.: #195001-8
- Stability under test conditions: Test material is considered stable at room temperature
- Specification: Calgon, Inc.

Test animals

Species:
rat
Strain:
other: Rochester (Ex-Wistar 1923)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: The original groups of rats were mated at 100 days
- Weight at study initiation: Bodyweight was approximately 70 grams for males and females at the start of the 100 day exposure.
- Fasting period before study:
- Housing: cages
- Diet (e.g. ad libitum): ad libitum,
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
no data

IN-LIFE DATES: From: *** To: ***

Administration / exposure

Route of administration:
oral: feed
Type of inhalation exposure (if applicable):
other: not applicable
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION

Diet mixtures were prepared using a basal ration of Purina Fox Chow Meal into which the appropriate amounts of sodium hexametaphosphate were mixed by a mechanical mixer. At weekly intervals. Diets were stored during the week in galvanized iron pails with covers.
Details on mating procedure:
- M/F ratio per cage: 1male: 2 female
- Length of cohabitation: 7 days
- Proof of pregnancy: no data
- Unsuccessful pairing replacement of first male by another male with proven fertility?: no.
- Further matings after two unsuccessful attempts: no data
- After successful mating each pregnant female was caged (how): no data
- other: males were rotated so that at each mating a different male would be placed in the cage with the female.

Details on mating schedules are tabulated below. - Table 1.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
not applicable
Duration of treatment / exposure:
Exposure took place for 100 days prior to mating, 21 days of gestation and during lactation for the P1, F1b, F2b. The F3b were exposed for their first 21days after which time they were sacrificed. 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 F1c and F3b litters were sacrificed after 21 days.
Frequency of treatment:
daily in feed
Details on study schedule:
- F1 parental animals were mated at 100 days of age in order to produce the sacrificial group F2a, and at 151 days F1 animals were mated to produce the next parental generation for F2b.
- Per female the litters were culled to ten pups with approximately 5/sex/group. There are no details on the selection of parental animals.
- Age at mating of the mated animals in the study: 100 days to produce the sacrifical litter and at 151 days to produce the next parental generation.

More details on protocol are tabulated below. - Table 1.
Doses / concentrations
Remarks:
Doses / Concentrations:
0.0% and 0.5%
Basis:
nominal in diet
No. of animals per sex per dose:
16 females and 8 males were used to produce the subsequent generation. P1 , F1b , F2b , F3b.
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Two doses were considered; 0.05% and 0.5%. After 100 days on these diets the 0.5% group exhibited no depression of body weight and was selected for use in this study.
- Other: The first litters born per generation were sacrificed, the F1a, F2a, F3a. The second litters born in a generation, the F1b, F2b, were mated to produce the next generation. The F1c was an additional litter produced and treated the same as the F1a.
Positive control:
no data

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: No

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were measured weekly during the 100 day pre-mating exposure for each generation.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Not measured

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not measured
Oestrous cyclicity (parental animals):
not evaluated
Sperm parameters (parental animals):
not evaluated
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 5 postpartum: yes
- excess pups were sacrificed to leave 10 per litter.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
[number of pups, pups/litter, pup mortality up to 21 days of age, weight gain per litter]

GROSS EXAMINATION OF DEAD PUPS: No
Postmortem examinations (parental animals):
SACRIFICE
- The first litters prodced per generation, the F1a, the F2a and the F3a were sacrificed at 30 days of birth.
- There is no data on the fate of the parents after they were mated to produce each generation.

GROSS NECROPSY
no data

HISTOPATHOLOGY
No histopathological examinations were performed for parental rats.

ORGAN WEIGHTS
No organ weight analyses were performed for parental rats.
Postmortem examinations (offspring):
SACRIFICE
- The non-parental litters, F1a , F1c, F2a , F3a , and F3b were sacrificed after 21 days.
- Observations amongst these litters were limited to litter weights on the 21st day. In the F3b histopathological and organ weight examinations were performed.

GROSS NECROPSY
F3b examined only.

HISTOPATHOLOGY / ORGAN WEIGTHS
The ten males and ten females from 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, lymph nodes, gut, marrow, pancreas) and organ weights (liver, kidneys, testes, lungs, brain, stomach, heart, spleen).
Statistics:
no data
Reproductive indices:
no data
Offspring viability indices:
no data

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
not specified
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed

Details on results (P0)

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)

In Table 2. a summary is presented of the reproductive performance.
Number of females mated: In all cases 15 or 16 females were mated after a 7-day cohabitation with one male (1 male caged with 2 females).
Number of pregnancies: Most of the pregnancies were successful. For the control rats there were 10 to 15 pregnancies per litter and for the rats maintained on the diet containing 0.5% HMP there were 10 to 13 in each mating.
Number of rats born: Comparable between the control and 0.5% group, the average number of pups ranged from 8-10 in both the controls and 0.5% group.

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
0.5 other: %
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed

Results: F1 generation

General toxicity (F1)

Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Details on results (F1)

VIABILITY (OFFSPRING)
The number of pups surviving at 21 days was 65-122 in the control groups and 69-111 in the 0.5% HMP group.
The average weight of the pups at 21 days was 41.8 - 48.7 g in the control groups and 40.9 - 45.4g in the 0.5% HMP group.

BODYWEIGHT (OFFSPRING)
After 21 days the newly born rats surviving to 21 days were weighed per litter, differences between the control and HMP-fed rats were not considered to be significant. See Table 2.

F1a - The average bodyweight of rats on the HMP diet was less than that of the control group.
F1b - The average bodyweight of rats on the HMP diet was less than that of the control group.
F1c - The average bodyweight of rats on the HMP diet was greater than that of the control group - based on this the authors considered the decreased average litter weights of the F1a and F1b to be mere coincidental variations.

F2a - The average bodyweight of rats on the HMP diet was less than that of the control group.
F2b - There was a small difference, less than a gram, in the average weight of the control and HMP-fed rats. There was a peculiar delay of the HMP-fed rats within the first week on the diet, but growth was rapidly recovered and surpassed that of the control group.

F3a - The average weight of the HMP-fed and control rats was almost identical at 42.5 and 42.4 gram respectively
F3b - The average weight of the MHP-fed rats was greater than that of the controls.

ORGAN WEIGHTS (OFFSPRING)
When the F3b rats were at weaning age they were sacrificed. The author described the organ weights and organ weight body weight ratios as, being within normal ranges. See Table 3

HISTOPATHOLOGY (OFFSPRING)
No histological changes were found that were attributed to the presence of HMP in the diet. No tumours were present and only normal tissues were found.

Effect levels (F1)

open allclose all
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 250 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
0.5 other: % w/w
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted

Target system / organ toxicity (F1)

Key result
Critical effects observed:
no

Results: F2 generation

General toxicity (F2)

Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not examined

Details on results (F2)

F2a - The average bodyweight of rats on the HMP diet was less than that of the control group.
F2b - There was a small difference, less than a gram, in the average weight of the control and HMP-fed rats. There was a peculiar delay of the HMP-fed rats within the first week on the diet, but growth was rapidly recovered and surpassed that of the control group.

Effect levels (F2)

Key result
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
0.5 other: %w/w
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted

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

Table 2. Reproductive performance

 Generation & Litter

F1a

F1b

F1c

F2a

F2b

F3a

F3b

 Controls

Females

16

15

15

16

15

16

16

Pregnancies

14

10

10

11

10

15

11

 

 

 

 

 

 

 

 

Total pups

107

95

77

94

93

145

115

Avg. pups/litter

7.6

9.5

9.6

8.5

9.3

9.7

10.4

 

 

 

 

 

 

 

 

Mortality (days 0-5)

0

1

3

0

10

8

10

Mortality (days 6-21)

0

1

5

3

4

9

6

 

 

 

 

 

 

 

 

Total rats surviving to day 21

107

84

65

87

77

122

90

Avg. weight of surviving rats

48.7

45.2

42.0

44.9

41.8

42.4

42.9

0.5% Hexametaphosphate group

Females

16

15

15

16

16

16

16

Pregnancies

11

10

13

13

10

12

10

 

 

 

 

 

 

 

 

Total pups

83

100

126

118

89

119

97

Avg. pups/litter

8.3

10.0

9.7

9.1

8.9

9.9

9.7

 

 

 

 

 

 

 

 

Mortality (days 0-5)

6

15

3

11

2

1

3

Mortality (days 6-21)

5

1

0

9

11

3

0

 

 

 

 

 

 

 

 

Total rats surviving to day 21

69

77

111

95

73

108

87

Avg. weight of surviving rats

43.1

43.5

44.7

44.3

40.9

42.5

45.4

Table 3 - Organ Weights

Group

No. of Rats

Body Wt.

Liver

Kidneys

Testes

Lungs

Brain

Stomach

Heart

Spleen

Control

Males

10

111

4.84

1.28

1.20

0.80

1.59

1.04

0.55

0.59

0.5%HMP-fed males

10

102

4.74

1.16

1.04

0.82

1.49

1.07

0.56

0.37

Control Females

10

96

4.03

1.06

n/a

0.69

1.43

1.02

0.53

0.64

0.5%HMP-fed females

10

90

3.71

1.03

n/a

0.67

1.54

1.09

0.47

0.36

Applicant's summary and conclusion

Conclusions:
Three generations of rats reared and maintained on diets containing 0.5% HMP showed no adverse effects to the test material - Control and test data were comparable across the range of observations made. Observations were limited to reproductive performance, pup mortality, organ weights and tissue and organ histopathology. Based on this evidence sodium metaphosphate is not considered to be classified as a reproductive toxicant according to Regulation (EC) 1272/2008 (EU CLP).

Although this study is limited and deficient by modern standards, it is still possible to make some valid scientific conclusions from the data. Adequate data are reported to show no effects on fertility or reproductive performance or on offspring growth and development over three generations with two litters per generation. Thus, the study should be considered acceptable for the dose level tested, 0.5% test material in the diet.