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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
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:
Developmental toxicity evaluation of 1,2,3,4 butanetetracarboxylic acid in Sprague Dawley (CD®) rats
Author:
Julia D. George, Catherine J. Price, Melissa C. Marr, Christina B. Myers,Gloria D. Jahnke
Year:
2001
Bibliographic source:
Reproductive Toxicology, 15, (2001), 413–420

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

Constituent 1
Chemical structure
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, NCAge 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 gFasting period before study:Housing:solid bottom, polycarbonate cages with stainless steel lidsUse of restrainers for preventing ingestion (if dermal): yes/no: No Diet (e.g. ad libitum): Purina Certified Rodent Chow, ad libitumWater (e.g. ad libitum): Durham, NC city water, ad libitumAcclimation period: 5 days quarantineENVIRONMENTAL CONDITIONSTemperature (°C): 70 to 73°FHumidity (%): 39 to 60%Air changes (per hr): Photoperiod (hrs dark / hrs light): 12 h light, 12 h darkIN-LIFE DATES: From: To: from gd 0 to gd 20

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
Details on exposurePREPARATION 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 availableVEHICLE- 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:1000.00mg/kg/day :25250.00mg/kg/day:25500.00mg/kg/day:251000.00mg/kg/day:25
Control animals:
yes, concurrent vehicle
Positive control:
No data available

Examinations

Parental animals: Observations and examinations:
Parental animals observation and examinationsCAGE SIDE OBSERVATIONS: Yes / No / No data: Not performedDETAILED CLINICAL OBSERVATIONS: Yes / No / No data: YesTime schedule: gd 0, 6, 9, 12, 15, 18, 19, and 20.BODY WEIGHT: Yes / No / No data: YesTime schedule for examinations: gd 0, 6, 9, 12, 15, 18, 19, and 20.FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): YesFood consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data: No data availableCompound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data availableWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data: YesTime 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 anesthetizedfetuses by evisceration
Postmortem examinations (parental animals):
SACRIFICEMale animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No dataMaternal 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 NECROPSYMaternal 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 exposureBody 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 20Test 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 availableReproductive function: sperm measures:No data availableReproductive 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 groupOrgan 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 availableHistopathology:No data availableother 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: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, non-treatment-related
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
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
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 availableBody 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 availableReproductive function: estrous cycle:No data availableReproductive function: sperm measures:No data availableReproductive performance:No data availableOrgan weights:No data availableGross 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 malformationsHistopathology:No data availableother findings:No data available

Effect levels (F1)

Dose descriptor:
other: not specified
Generation:
other: not specified
Based on:
not specified
Sex:
not specified
Basis for effect level:
other: not specified
Remarks on result:
other: not specified

Results: F2 generation

General toxicity (F2)

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 (F2)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not specified

Overall reproductive toxicity

Reproductive effects observed:
no
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
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.

 

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