Dihydrogen [[N,N'-ethylenebis[N-(carboxymethyl)glycinato]](4-)-N,N',O,O',ON,ON']nickelate(2-)

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Evaluation of the given test chemical in Sprague Dawley (CD®) rats.

Author:

George et al.

Year:

2001

Bibliographic source:

Reproductive Toxicology

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: under a protocol approved by the RTI Institutional Animal Care and Use Committee (IACUC)

Principles of method if other than guideline:

Reproductive and developmental toxicity study of 1,2,3,4 butane tetra carboxylic acid was performed on Sprague Dawley (CD®) rats

GLP compliance:

not specified

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Butane-1,2,3,4-tetracarboxylic acid

EC Number:

216-938-0

EC Name:

Butane-1,2,3,4-tetracarboxylic acid

Cas Number:

1703-58-8

Molecular formula:

C8H10O8

IUPAC Name:

Butane-1,2,3,4-tetracarboxylic acid

Test material form:

solid

Details on test material:

- Name of the test chemical: Butane-1,2,3,4-tetracarboxylic acid
- IUPAC name: Butane-1,2,3,4-tetracarboxylic acid
- Molecular Formula: C8H10O8
- Molecular Weight: 234.159 g/mol
- Smile Notation: OC(=O)C[C@H]([C@H](CC(=O)O)C(=O)O)C(=O)O
- InChI : 1S/C8H10O8/c9-5(10)1-3(7(13)14)4(8(15)16)2-6(11)12/h3-4H,1-2H2,(H,9,10)(H,11,12)(H,13,14)(H,15,16)/t3-,4+
- Substance type: organic

Specific details on test material used for the study:

- Name of the test chemical: Butane-1,2,3,4-tetracarboxylic acid
- IUPAC name: Butane-1,2,3,4-tetracarboxylic acid
- Molecular Formula: C8H10O8
- Molecular Weight: 234.159 g/mol
- Smile Notation: OC(=O)C[C@H]([C@H](CC(=O)O)C(=O)O)C(=O)O
- InChI : 1S/C8H10O8/c9-5(10)1-3(7(13)14)4(8(15)16)2-6(11)12/h3-4H,1-2H2,(H,9,10)(H,11,12)(H,13,14)(H,15,16)/t3-,4+
- Substance type: organic

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Source: Charles River Laboratories, Inc., Raleigh, NC
Age at study initiation: (P) x wks; (F1) x wks: (Parent) 8 to 10 weeks old;
Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: Females (P)- 218 to 269 g
Fasting period before study:
Housing:solid bottom, polycarbonate cages with stainless steel lids
Use of restrainers for preventing ingestion (if dermal): yes/no: No
Diet (e.g. ad libitum): Purina Certified Rodent Chow, ad libitum
Water (e.g. ad libitum): Durham, NC city water, ad libitum
Acclimation period: 5 days quarantine

ENVIRONMENTAL CONDITIONS
Temperature (°C): 70 to 73°F
Humidity (%): 39 to 60%
Air changes (per hr):
Photoperiod (hrs dark / hrs light): 12 h light, 12 h dark

IN-LIFE DATES: From: To: from gd 0 to gd 20

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0, 250, 500, and 1000 mg/kg/day
- Amount of vehicle (if gavage): 5 ml/kg
- Lot/batch no. (if required): No data available
- Purity: >99%

Details on mating procedure:

- M/F ratio per cage:1:1
- Length of cohabitation:After quarantine individual females were placed overnight in the home cage of a singly housed male of the same stock for mating
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:presence of sperm in the vaginal lavage
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.:No data available

- Further matings after two unsuccessful attempts: [no / yes (explain)]:No data available

- After successful mating each pregnant female was caged (how):No data available

- Any other deviations from standard protocol:No data available

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Dosing solutions (BTCA in deionized/distilled water) were verified to be within 92 to 103% of their theoretical concentrations by HPLC prior to and after the period of administration

Duration of treatment / exposure:

14 days (Gd 6 to Gd 20)

Frequency of treatment:

Gd 0, 6, 9, 12, 15, 18, 19, and 20.

Details on study schedule:

No data available

Doses / concentrations

Remarks:

0,250,500,1000 mg/kg/day

No. of animals per sex per dose:

Total:100
0.00mg/kg/day :25
250.00mg/kg/day:25
500.00mg/kg/day:25
1000.00mg/kg/day:25

Control animals:

yes, concurrent vehicle

Positive control:

No data available

Examinations

Parental animals: Observations and examinations:

Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: Yes / No / No data: Not performed

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data: Yes
Time schedule: gd 0, 6, 9, 12, 15, 18, 19, and 20.
BODY WEIGHT: Yes / No / No data: Yes
Time schedule for examinations: gd 0, 6, 9, 12, 15, 18, 19, and 20.

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


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data: Yes
Time schedule for examinations: gd 0, 6, 9, 12, 15, 18, 19, and 20.

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

Each anesthetized fetus was counted, weighed, and examined for external morphologic abnormalities, including cleft palate Approximately one-half (50%) of the anesthetized fetuses were terminated by decapitation and the remaining anesthetized
fetuses by evisceration

Postmortem examinations (parental animals):

SACRIFICE
Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No data
Maternal animals: yes
All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]: Following termination by CO2 asphyxiation on gd 20,

GROSS NECROPSY
Maternal liver weight and gravid uterine weight were measured and corpora lutea were counted. Uterine contents were evaluated for the number of implantation sites, resorptions, late fetal deaths (i.e. fetuses with discernible digits and weighing greater than 0.9 g, but displaying no vital signs at the time of uterine dissection), and live fetuses. The uterus was stained to reveal possible early resorptions

Postmortem examinations (offspring):

No data available

Statistics:

The unit for statistical measurement was the pregnant female or the litter. Quantitative continuous data were compared among treatment groups by parametric tests if Bartlett’s test for homogeneity of variance was not significant. If Bartlett’s test indicated a lack of homogeneity (p , 0.001), then nonparametric tests were applied. Statistical analyses were based on SAS software. General Linear Models (GLM) were applied to the Analyses of Variance (ANOVA) and the Tests for Linear Trend. Prior to GLM analysis, an arcsine-square root transformation was performed on all litter-derived percentage data.
For litter-derived percentage data, the ANOVA was weighted according to litter size. If a significant (p , 0.05) main effect for dose occurred, then Dunnett’s Multiple Comparison Test was used to compare each treatment group to the control group for that measure.
A two-tailed Dunnett’s Test was used for maternal body and organ weight parameters, maternal feed and water consumption, fetal body weight, and percent male fetuses per litter.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Details on results (P0)

Clinical signs:Treatment-related clinical signs included weight loss (≤5 g in any one weigh period), which occurred occasionally in all groups and showed increasing frequency and/or severity with increasing dose on individual days during the treatment period). Piloerection was noted in ≤4 females/day at the high dose. Rooting in the cage bedding after dosing suggested an aversion to the sensory properties of the mid- and high-dose formulations (≤3 mid-dose females/day and ≤5 high-dose females/day). Alopecia was noted with a low incidence in all groups but showed no apparent relationship to BTCA exposure

Body weight and food consumption:Maternal body weight was comparable among groups on gd 0 and 6 (i.e. prior to the initiation of dosing). On gd 9, 12, 15, 18, and 19, maternal body weights at the low and mid doses were not affected (98.8 to 100.8% of concurrent controls), but maternal body weight at the high dose was significantly lower than the vehicle control on each of these days, except gd 9. On gd 20, maternal body weight at the high dose was significantly below the vehicle control. Decreases in maternal body weight became larger with repeated dosing, and the maximum reductions occurred at the end of the dosing period (gd 18, 19, and 20

Test substance intake:Maternal relative feed intake was transiently reduced at 1000 mg/kg/day from gd 6 to 9 and at 500 mg/kg/day from gd 12 to 15, although no changes were noted for the treatment period as a whole. Maternal relative water intake was increased at 1000 mg/kg/day for gd 12 to 15, 18 to 19, 19 to 20, and the treatment period as a whole.

Reproductive function: estrous cycle:No data available

Reproductive function: sperm measures:No data available

Reproductive performance:Treatment-related maternal mortality occurred in 4% (1/24) of high-dose females. All remaining females survived until scheduled necropsy on gd 20; pregnancy was confirmed in 84 to 100% per group

Organ weights:Gravid uterine weight and maternal relative liver weight were not affected. Absolute maternal liver weight exhibited a decreasing trend but showed no significant differences between control and BTCA-treated groups

Gross pathology:No data available

Histopathology:No data available
other findings:No data available

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

body weight and weight gain

food consumption and compound intake

haematology

Remarks on result:

other: not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Details on results (F1)

Clinical signs:No data available

Body weight and food consumption:Average fetal body weight per litter in BTCA-treated groups was comparable to controls (97 to 103% of the average control weight for males, females or both sexes combined), and no significant dose-related trends were found.

Test substance intake:No data available
Reproductive function: estrous cycle:No data available

Reproductive function: sperm measures:No data available

Reproductive performance:No data available

Organ weights:No data available
Gross pathology:External malformations were limited to one control fetus with a short tail. Visceral malformations included hydronephrosis and/or hydroureter (3, 0, 5, and 1 fetuses in the control through high-dose groups). Each of these visceral findings has a recognizable background incidence in this species/strain and the incidence in this study failed to show a dose-related pattern. Skeletal malformations
(fused ribs or centra and split centra) were found in only one fetus in the control group and two fetuses in the Low - dose group. Thus, no association was found between BTCA exposure and the incidence of malformations

Histopathology:No data available

other findings:No data available

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

500 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: NOAEL was found to be 500mg/kg bw/day

Target system / organ toxicity (F1)

Critical effects observed:

no

Overall reproductive toxicity

Reproductive effects observed:

no

Treatment related:

not specified

Applicant's summary and conclusion

Conclusions:

The reproductive toxicity study NOAEL considered to be 500 mg/kg/day, When CD rats were treated with 1,2,3,4-butanetetracarboxylic acid (BTCA) orally.

Executive summary:

1,2,3,4-butanetetracarboxylic acid (BTCA) was tested for reproductive toxicity.The experimental animals were Sprague Dawley of 8 to 10 weeks old. After quarantine, individual females were placed overnight in the home cage of a singly housed male of the same stock for mating, and then examined the next morning for the presence of sperm in thevaginal lavage. Female rats weighed from 218 to 269 gon gd 0 (i.e. day of vaginal sperm detection). Time-matedfemales were assigned to treatment groups (25/group) bystratified randomization for body weight on gd 0.

 

The BTCA doses chosen for this study were 0, 250, 500, and 1000 mg/kg/day administered by gavage (5 ml/kg). Dose range selection for this study was based in part on an acute, oral toxicity study of BTCA [800, 1260, 1590, 2000, and 3170 mg/kg] in COX-SD rats. Dose range selection was also supported by a study in non-pregnant female CD rats (7 per group) exposed to BTCA [0, 50,100,500,750, and 1000 mg/kg/day] for 14 consecutive days.

 

Following termination by CO₂asphyxiation on gd 20, maternal liver weight and gravid uterine weight were measured and corpora lutea were counted. Uterine contents were evaluated for the number of implantation sites, resorptions, late fetal deaths (i.e. fetuses with discernible digits and weighing greater than 0.9 g, but displaying no vital signs at the time of uterine dissection), and live fetuses. The uterus was stained to reveal possible early resorptions.

 

Maternal feed and water consumption, body weight, and clinical signs were monitored throughout gestation. At termination (gd 20), confirmed-pregnant females (21 to 25 per group) were evaluated for clinical status and gestational outcome; live fetuses were examined for external, visceral, and skeletal malformations. There were no treatment-related effects on fetal growth, survival, or morphologic development. Therefore NOAEL considered to be 500 mg/kg/day, for the reproductive toxicity study of1, 2,3,4-butanetetracarboxylic acid (BTCA) was performed on CD rats.