[ethane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

281 days overall

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

This study was conducted to assess the long-term effects of CP 41863 when administered continuously via the diet to Long-Evans rats at concentrations of 300, 1000 and 3000 ppm through one complete generation. Test substance administration to the F0 generation females (201group) was initiated at the onset of gestation and continued throughout the ensuing gestation and lactation periods. Dietary administration of CP 41863 continued to the F1 generation animals (10 males and 20 females/group) through a growth period and mating, gestation and lactation period for two successive litters.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

[ethane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid

EC Number:

215-851-5

EC Name:

[ethane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid

Cas Number:

1429-50-1

Molecular formula:

C6H20N2O12P4

IUPAC Name:

[ethane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid

Constituent 2

Reference substance name:

Ethylenediaminetetra (methylene phosphonic acid)

IUPAC Name:

Ethylenediaminetetra (methylene phosphonic acid)

Test material form:

not specified

Test animals

Species:

rat

Strain:

Long-Evans

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: B1 ue Spruce Farms
A1 tamont, N.Y.
- Age at study initiation: (P) 15-16 weeks
- Housing:Individually in elevated stainless
steel cages (except during mating).
- Diet (e.g. ad libitum):Standard laboratory diet (Purina
Laboratory Chow@ 5001 )
-ad l ibitum.
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Temperature
monitored twice daily
- Humidity (%): Not stated
- Air changes (per hr):Not stated
- Photoperiod (hrs dark / hrs light):12 hour l i ght/dark cycle.

Administration / exposure

Route of administration:

oral: feed

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:No information


- Lot/batch no. (if required):176-18-8-B
- Purity:93.3%

Details on mating procedure:

F0 Females: Females were placed with male rats nightly in a ratio of 2:l. Vaginal smears were taken early in the morning and females were considered t o have mated if sperm and/or vaginal plug was observed. The day on which evidence of mating was observed was defined as Day 0 of gestation.

F1 Females: One male and two females of equivalent dose levels were caged together nightly for 15 nights or until a sign of mating (sperm and/or vaginal plug) was observed. Care was taken t o avoid brother-sister mating. The day on which evidence of mating was observed was defined as Day 0 of gestation.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Test substance was administered continuously , beginning Day 0 of gestation for the F0 females and continued for the F1 males and females through the F2a and F2b l itters. Age of F0 and initiation of test substance administration: 15-16 weeks.

Frequency of treatment:

Daily

Details on study schedule:

Test substance administration of the F0 females was initiated the day signs of mating were observed (Day 0 of gestation) and continued throughout the ensuing gestation and l actation periods. F1 offspring were separated from siblings seven days after the weaning (Day 21 of lactation) of the last litter and were randomly selected to continue as future parents ( F1 ).

F1 anmals were raised to maturity and mated to produce the F2, litters. F2a pups were sacrificed, necropsied and discarded at weaning. All F1 females were remated after a rest period of at least 14 days to produce the F2b litters . F2b pups were sacrificed and necropsied at weaning.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 ppm

Remarks:

Group 1

Doses / concentrations 2

Dose / conc.:

300 ppm

Remarks:

Group 2

Doses / concentrations 3

Dose / conc.:

1 000 ppm

Remarks:

Group 3

Doses / concentrations 4

Dose / conc.:

3 000 ppm

Remarks:

Group 4

No. of animals per sex per dose:

Fo - (females) - 20/group
F1 - (males and females) - 10 M + 20 F /group

Control animals:

yes

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
Physical observations -twice daily.For mortality and gross signs of toxicologic or pharmacologic effects. (Fo, F1 parents)
Detailed physical observations - for signs of local or systemic toxicity, pharrnacologic effects and palpation for tissue masses (Fo, F1 parents) :

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

BODY WEIGHT: Yes
- Mean maternal body weight during gestation and lactation; mean pup body weight recorded atdays 0,4,14,21 of lactation; mean body weight recorded during F1 growth and rest periods.

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

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

> 3 000 mg/kg bw (total dose)

Based on:

labile/free

Remarks:

acid

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

no effects observed

Results: F1 generation

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

> 3 000 ppm

Based on:

labile/free

Remarks:

acid

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity (F1)

Critical effects observed:

no

Results: F2 generation

Effect levels (F2)

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

> 3 000 ppm

Based on:

other: labile/free

Remarks:

Acid

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

F0 generation - No treatment related effects were evident in respect to growth, food consumption, reproductive-fertility, or litter examination parameters. At terminal sacrifice the mid dose males and high dose animals had lower body weights. In the mid and high dose levels males had higher adrenal weights both absolute and relative to body and brain weights. High dose females had higher thyroid/body and spleen/body weight ratios. However organ to brain weight ratios revealed no treatment effect in the F1 females.

Gross postmortem observations and evaluation of selected tissues from five adult F1 generation males and females of the control (Groups I and II) and high dose (Group V) groups indicated no treatment related effects.

No treatment related effects on any of the reproductive parameters. (Mating, fertility; pregnancy, parturition & survival)

Applicant's summary and conclusion

Executive summary:

In a one generation reproductive toxicity study the test substance was administered continuously via the diet to Long-Evans F0 generation females (20/group) at the onset of gestation and continued throughout the ensuing gestation and lactation periods. Dietary administration was continued to the F1 generation animals (10 males and 20 females /group) through a growth period and a mating, gestation and lactation period for two successive litters.

In the F0 generation, no treatment effects were evident in the low or mid dose groups. In the F1 generation, no treatment effects were observed in growth, food consumption, fertility-reproduction or litter examination parameters. At terminal sacrifice mid dose males an high dose animals had lower body weights. In the mid and high dose levels males had higher adrenal weights, both absolute and relative to body and brain ratios.

However in the absence of histopathological/clinical pathology examination results, it is not clear whether or not the weight change a real effect. In the absence of adverse clinical /reproductive effects for this substance it is unlikely to be of toxicological significance.

Therefore the NOAEL for reproductive effects for F0 and F1 is > 3000 ppm.