Registration Dossier
Registration Dossier
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 416-530-4 | CAS number: 178949-82-1
Endpoint summary
Administrative data
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Effects on fertility
Description of key information
In a GLP study conducted according to OECD
Guideline 415, no adverse affects on measures of fertility and
development (or systemic toxicity) were seen in groups of rats
administered trisodium EDDS by stomach tube at up to 700 mg/kg bw/day
(considered the study NOAEL) in a one-generation reproduction study
(York, 1999).
No 2-, 3- or multi-generation studies on trisodium EDDS, EDDS (or its
other simple salts), or EDTA were identified.
However, data from a multigeneration study on rats with calcium disodium
EDTA gave no evidence for adverse effects on reproductive performance
and outcome at up to 250 mg/kg bw/day, considered the study NOAEL (Oser
et al. 1963). In a limited and briefly reported 2-generation
reproductive toxicity study, normal first and second litters were
reported from rats fed disodium EDTA in the diet at up to about 500
mg/kg bw/day (considered the study NOAEL), while those animals of the
highest dose (2500 mg/kg bw/day) failed to produce any litters. Data on
the second generation were not available (Yang, 1952).
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), trisodium EDDS caused a dose-related and statistically significant reduction in plasma levels of copper and zinc, but no such reductions in plasma iron levels were seen.
Maternal toxicity, evident as soft faeces and a slight reduction in body weight gain, was seen at 1000 mg/kg bw/day (the highest tested dose) and this is considered in this study as the NOAEL.
Link to relevant study records
- Endpoint:
- one-generation reproductive toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 14 September 1998 to 20 January 1999
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
- Deviations:
- no
- GLP compliance:
- yes
- Limit test:
- no
- 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., Portage, Michigan
- Age at study initiation: (P) 7 wks
- Weight at study initiation: (P) Males: 106-132 g; Females: 105-132 g
- Fasting period before study: no data
- Housing: individually, except during mating and postpartum period, in stainless steel, wire-bottomed cages
- Diet (e.g. ad libitum): conventional with mineral supplement containing twice the National Research Council recommended daily intake of copper, iron and zinc (as EDDS acid has been shown to chelate these minerals in previous studies), ad libitum
- Water (e.g. ad libitum): chlorinated, reverse osmosis membrane processed, deionised water
- Acclimation period: 6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- oral: gavage
- Vehicle:
- water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: prepared weekly with continuous stirring at concentrations of 9, 25 and 70 mg/mL
- Details on mating procedure:
- - M/F ratio per cage: 1:1
- Length of cohabitation: up to 14 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility for a further 7 days
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually with nesting material
- Any other deviations from standard protocol: no - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- No data. [Duplicate samples were taken from the top, middle and bottom of each concentration on first day of dosing and then monthly and sent away for analysis.]
- Duration of treatment / exposure:
- From 70 days before mating until postpartum day 21
- Frequency of treatment:
- Once daily
- Details on study schedule:
- Age at mating of the mated animals in the study: 18 weeks
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 90 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 250 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 700 mg/kg bw/day (actual dose received)
- No. of animals per sex per dose:
- 25/sex/dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: doses determined by the sponsor
- Rationale for animal assignment (if not random): computer-generated (weight-ordered) randomization - Positive control:
- none
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: yes
- Time schedule: twice daily
- Cage side observations included: deaths, and in females behaviour, abortions and premature deliveries.
DETAILED CLINICAL OBSERVATIONS: yes
- Time schedule: daily
BODY WEIGHT: yes
- Time schedule for examinations: males - on day 1 of dosing, then weekly. Females - on day 1 of dosing, then weekly until gestation, then on gestation days 0, 6, 10, 15 and 25 (if necessary) and on lactation days 1,4, 7, 10, 14 and 21.
FOOD CONSUMPTION: yes
- Time schedule: males - weekly during dosing period. Females - weekly until gestation, then on gestation days 0, 6, 10, 15 and 25 (if necessary) and on lactation days 1,4, 7, 10 and 14. - Oestrous cyclicity (parental animals):
- Estrous cycle observed for 14 days prior to cohabitation
- Sperm parameters (parental animals):
- Parameters examined in male parental generations:
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:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were sacrificed and not examined further.
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities
GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead. - Postmortem examinations (parental animals):
- SACRIFICE
- Male animals: all surviving animals were evidently sacrificed after weaning (although the methods section of the report states males were sacrificed once pregnancy was confirmed)
- Maternal animals: all surviving animals were sacrificed after their litter was weaned. Females that did not deliver a litter were sacrificed on gestation day 25.
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the pelvic, thoracic and abdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for possible future microscopic examination and weighed: prostate, pituitary and brain; uterus and ovaries (of females); epididymides, seminal vesicles and testes (of males). All gross lesions were retained and those from dams were examined histologically. Histopathology was performed on the testes of all control and high dose males, and any males that failed to induce pregnancy. Micropscopic examination of the ovaries of all females that were not pregnant was also carried out.
OTHER EXAMINATIONS
Baseline serum levels of copper, iron and zinc in 5 animals/sex were taken prior to treatment. Blood was collected from females at 40 days of age and terminal sacrifice and from males on day of sacrifice and the serum analysed for copper, iron and zinc levels. - Postmortem examinations (offspring):
- SACRIFICE of pups kept until weaned
- The F1 offspring were sacrificed at 21 days of age
- These animals were not subjected to postmortem examinations.
GROSS NECROPSY
None
HISTOPATHOLOGY / ORGAN WEIGTHS
None
SACRIFICE of pups culled due to standardization
- The F1 offspring were sacrificed at 4 days of age
- These animals were subjected to postmortem examinations.
GROSS NECROPSY
Gross necropsy consisted of external and internal examinations, including the pelvic, thoracic, and abdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGTHS
Cross-section of the brain at the level of the frontal-parietal suture for hydrocephaly. - Statistics:
- Bartlett's test of homogeneity of variances. If the Bartlett's test was not significant, then the analysis of variance was applied and if this was significant, Dunnett's test was used to identify the statistical significance of the individual groups. If the Bartlett's test was significant, the Kruskal-Wallis test or Fisher's exact test were applied as appropriate.
Discrete natural delivery data were evaluated using the Kruskal-Wallis test. - Reproductive indices:
- Precoital index (days in cohabitation before mating), mating index, fertility index, gestation index
- Offspring viability indices:
- Number and sex of offspring/litter, number of implantation sites, condition of pups, litter size and viability, number of pups surviving to lactation days 1 and 4, lactation index (number of pups surviving to weaning), percentage survival and sex ratio.
- Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Description (incidence):
- There were no treatment-related deaths or clinical signs reported
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- A slight (about 5%), but not statistically significant, reduction in body weight gain was observed in the high-dose males (throughout the study) and females (prior to mating only), compared to the controls
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- Food consumption was slightly reduced (not statistically significant) in males at the top-dose level throughout the study.
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- not examined
- Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- The zinc levels in serum were elevated in males in all treatment groups and in females only at the top dose. There were no changes in the serum levels of copper and iron.
- Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- No effects were seen at any stage of the estrous cycle.
- Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- No differences were seen in sperm number, motility, cauda epididymal sperm count or density.
- Reproductive performance:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The precoital, mating and fertility indices were similar in all groups. The number of implantations and litters, lactation index and sex ratios of the pups were similar between groups. A slight, non dose-related non-significant, reduction in the number of pregnant dams was seen; 23 (95.8%), 22 (88.0%), 22 (91.7%) and 20 (80.0%) pregnant females in the 0, 90, 250 and 700 mg/kg bw/day groups, respectively.
- Key result
- Dose descriptor:
- NOAEL
- Remarks:
- parental toxicity
- Effect level:
- ca. 700 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Remarks:
- No treatment related effects at highest dose
- Key result
- Dose descriptor:
- NOAEL
- Remarks:
- fertility
- Effect level:
- ca. 700 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Remarks:
- No treatment related effects at highest dose
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No effects were attributed to the test substance. In all groups, persistent clinical effects, each occurring in 1 pup/group consisted of tip of tail missing, microphthalmia, bent tail and umbilical hernia.
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- Viability was similar in all groups.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- No significant differences were seen between groups.
- Sexual maturation:
- not examined
- Organ weight findings including organ / body weight ratios:
- not specified
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- No significant differences were seen between the groups.
- Histopathological findings:
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- ca. 700 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Remarks:
- No treatment related effects at highest dose
- Reproductive effects observed:
- not specified
- Conclusions:
- In a GLP study conducted according to OECD Guideline 415, no adverse affects on measures of fertility and development (or systemic toxicity) were seen in groups of rats administered trisodium EDDS by stomach tube at up to 700 mg/kg bw/day (considered the study NOAEL) in a one-generation reproduction study.
- Executive summary:
In a GLP study conducted according to OECD Guideline 415, trisodium EDDS was assessed for its ability to induce reproductive and developmental toxicity in rats.
Groups of 25 Spague-Dawley rats of each sex were given 0, 90, 250 or 700 mg/kg bw/day of the test substance (in water) by oral gavage for 70 days before mating, and throughout mating, pregnancy and weaning. Culling of the offspring occurred on day 4 after birth to standardize the litters, and the remaining offspring were killed on day 21 after delivery. The animals were observed for mortality, clinical signs of toxicity, body weight gain, feed consumption, changes in the estrus cycle, precoital index, mating index, fertility, gestation index, number and sex of offspring, litter size and viability, and lactation index. At sacrifice, gross necropsy of the thoracic, abdominal and pelvic viscera was carried out, the reproductive organs, brain and pituitary were weighed and sperm evaluations were performed. The females were examined for the number and distribution of implantation sites. Necropsies were performed on the pups culled at 4 days for external and internal abnormalities and the brains were sectioned to detect hydrocephaly.
No significant systemic adverse effects were observed on the parental animals, and the reproductive and developmental parameters examined were unremarkable. Therefore the study NOAEL was considered to be 700 mg/kg bw/day.
In conclusion, trisodium EDDS did not affect fertility or development in rats orally administered up to 700 mg/kg bw/day in a one-generation reproductive toxicity study. Therefore, according to EU CLP regulation, trisodium EDDS would not be classified as a reproductive or developmental toxicant under the test conditions.
- Endpoint:
- multi-generation reproductive toxicity
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- no guideline available
- Principles of method if other than guideline:
- In a multigeneration rat study, the parental (P) generation were fed the test substance from weaning and at sexual maturity underwent two successive matings. Exposure was continued throughout the study. Ten rats of each sex were selected from as many litters as possible from successive generations, and assigned to the F1, F2 and F3 generation, respectively, under the same exposure and mating regime as the P generation. The animals were observed for effects on fertility, gestation and lactation and for viability of the offspring. The P generation were exposed for 2 years, after which time the study was terminated (i.e. the F1, F2 and F3 generations were exposed for 18, 12 and 6 months, respectively). At study termination, the animals were examined for gross abnormalities and 15 organs and tissues were examined microscopically.
- GLP compliance:
- no
- Remarks:
- prior to GLP
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Food and Drug Research Laboratories Inc., Maspeth 78, New York, USA
- Age at study initiation: at weaning (21 days)
- Weight at study initiation: no data given at start of exposure. At 12 weeks: (P) Males: 259-285 g; Females: 146-157 g; (F1) Males: 259-312 g; Females: 135-159 g; (F2) Males: 232-286 g; Females: 134-168 g; (F3) Males: 280-306 g; Females: 152-173 g
- Fasting period before study: no data
- Housing: individually (apart from during mating and lactation)
- Diet (e.g. ad libitum): "natural foods supplemented with inorganic salts and vitamins". Reported to "resemble the food consumption pattern of the US population with respect to the ratios of milk, meat and grain components"; ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data - Route of administration:
- oral: feed
- Vehicle:
- not specified
- Details on exposure:
- DIET PREPARATION
- Rate of preparation of diet (frequency): no data
- Mixing appropriate amounts with (Type of food): no data
- Storage temperature of food: no data
- Concentration: 25% solution - Details on mating procedure:
- - M/F ratio per cage: 1 male to 2 females
- Length of cohabitation: up to 3 weeks
- Proof of pregnancy: visually, by palpation or by weight increments. Not referred to by day of pregnancy
- After 21 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no, the female was regarded as infertile and matings discontinued
- After successful mating each pregnant female was caged: individually - Analytical verification of doses or concentrations:
- not specified
- Details on analytical verification of doses or concentrations:
- no data
- Duration of treatment / exposure:
- From weaning until 6, 12, 18, or 24 months after start of exposure for the F3, F2, F1 or P generations, respectively.
- Frequency of treatment:
- Daily in diet; ad libitum
- Details on study schedule:
- - F1, F2 and F3 parental animals not mated until 14 weeks after selection from the litters. A second mating took place one week after weaning of the offspring
- Selection of parents from F1, F2 and F3 generation when pups were 21 days of age.
- Age at mating of the mated animals in the study: 13 and 21 weeks (2 litters) - Dose / conc.:
- 20 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 125 mg/kg bw/day (nominal)
- Dose / conc.:
- 250 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 25/sex/dose in the P generation, then 10 rats/sex/dose in the F1, F2 and F3 generations
- Control animals:
- yes, plain diet
- Details on study design:
- - Dose selection rationale: based on a range-finding study
- Rationale for animal assignment (if not random): random - Positive control:
- No
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations for physical condition and behaviour
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: no details; "for the 12-week postweaning period"
BODY WEIGHT: Yes
- Time schedule for examinations: weekly
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: food consumption for each of 10 rats/sex recorded as a representative sample of the group
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: no data - Oestrous cyclicity (parental animals):
- No data
- Sperm parameters (parental animals):
- Not examined
- Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no
10 rats/sex were selected at 21 days (at weaning) to provide the successive generations. The P generation were kept until they reached 2 years of age when the study was terminated (i.e. the F1, F2 and F3 generations were kept for 18, 12 and 6 months, respectively)
PARAMETERS EXAMINED
The following parameters were examined in F1 / F2 / F3 offspring:
viability and lactation index, postnatal mortality, weight gain, external and internal examinations, organ weights, physical or behavioural abnormalities
GROSS EXAMINATION OF DEAD PUPS:
yes, the main organs, including gonads were examined - Postmortem examinations (parental animals):
- SACRIFICE
- Male and maternal animals: 2 animals/sex/group were sacrificed at 12 weeks. All surviving animals in the 50 and 125 mg/kg bw/day groups not selected to provide the next generation were sacrificed after weaning of the second litters. Control animals and animals in the high-dose group continued on the diet for a total of 2 years.
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated were prepared for microscopic examination and weighed, respectively: in animals sacrified at 12 weeks; liver, kidneys, spleen, heart, adrenals, thyroid and gonads. At study termination, kidneys, pancreas, heart, spleen, lungs, bone marrow, stomach, small and large intestines, gonads, thyroid, parathyroids, lymph nodes, spinal cord, and tibia.
At 6 and 12 weeks from start of exposure in about one quarter of the animals/sex/group the following parameters were determined: blood haemoglobin levels, red and white blood cell counts, differential white cell counts, prothrombin time, blood sugar and non-protein nitrogen, serum calcium, urinary albumin and sugar.
Due to the sequestering action of calcium disodium EDTA which may interfere with mineral metabolism, the following additional examinations were made: ash content of tibias, examination for dental caries, xanthine oxidase in liver and carbonic anhydrase in serum. - Postmortem examinations (offspring):
- SACRIFICE
The F1 and F2 offspring not selected as parental animals were sacrificed after weaning of the second litters. The F3 offspring were sacrificed at birth. The F1, F2 and F3 parental animals were sacrificed at 18, 12 and 6 months, respectively.
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera
HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated were prepared for microscopic examination and weighed, respectively: kidney, pancreas, heart, spleen, lungs, bone marrow, stomach, small and large intestines, gonads, thyroid, parathyroid, lymph nodes, spinal cord, and tibia. - Statistics:
- No data
- Reproductive indices:
- Fertility index, gestation index
- Offspring viability indices:
- Viability index (the proportion of offspring that survived 4 days or longer); lactation index (the proportion of offspring alive at 4 days that survived to weaning)
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No clinical signs of toxicity were evident
- Mortality:
- no mortality observed
- Description (incidence):
- No treatment-related deaths were seen. In the P generation, 3 deaths occurred in control animals at 10-12 weeks, 1 death occurred at 12 weeks at 50 mg/kg bw/day and 1 death occurred at 8 weeks at 125 mg/kg bw/day.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Body weights were similar in all groups
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- Food consumption was similar in all groups
Dietry consumption, and therefore substance intake, were comparable between groups - Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- No significant differences were evident between the groups
- Other effects:
- no effects observed
- Description (incidence and severity):
- There were no significant differences in the levels of haemoglobin, haematocrit, blood sugar and non-protein nitrogen, urinary albumin and sugar, red and white blood cell counts, differential white cell counts and prothrombin times.significant differences were detected in the extent of dental caries in the high-dose and control groups.
The activity of the two metallo-enzymes, carbonic anhydrase and xanthine oxidase, were similar in all groups. - Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- No significant differences in reproductive performance were evident between groups
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- ca. 250 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Remarks:
- No effects were seen on clinical signs; mortality; body weight; food consumption and compound intake; food efficiency; water consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; birth index; live birth index; pregnancy index; litter size; litter weight; pup weight; survival index; viability index; lactation index
- Key result
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No clinical signs of toxicity were evident and no deaths occurred in the F1, F2 or F3 generations
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- No significant differences were seen in viability between groups in the F1, F2 or F3 generations
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Body weights and food consumption were similar in all groups in all generations
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- Diet consumption, and therefore substance intake, was comparable between groups in all generations
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- There were no significant differences in the levels of haemoglobin, haematocrit, blood sugar and non-protein nitrogen, urinary albumin and sugar, red and white blood cell counts, differential white cell counts and prothrombin times.
- Sexual maturation:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- Organ weights were similar in all groups
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- No significant differences were evident between the groups
- Histopathological findings:
- no effects observed
- Description (incidence and severity):
- No significant differences were evident between the groups
- Other effects:
- no effects observed
- Description (incidence and severity):
- REPRODUCTIVE PERFORMANCE (OFFSPRING)
No significant differences in reproductive performance were evident between groups
No significant differences were detected in the extent of dental caries in the high-dose and control groups.
The activity of the two metallo-enzymes, carbonic anhydrase and xanthine oxidase, were similar in all groups. - Key result
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Reproductive effects observed:
- not specified
- Conclusions:
- In a good quality study (prior to GLP), no adverse effects were evident in a multigeneration reproduction study in rats fed calcium disodium EDTA at up to 250 mg/kg bw/day in the diet, considered the study NOAEL.
- Executive summary:
In a good-quality study (prior to GLP), calcium disodium EDTA dihydrate was assessed for the potential to cause adverse effects in a multigeneration reproductive toxicity study in Wistar rats.
Groups of 25 rats of each sex (P generation) were fed 0, 50, 125 or 250 mg/kg bw/day in the diet from weaning (day 21) for up to 2 years. At 13 and 21 weeks of age, male rats were mated with females to provide two litters (F1 generation). At weaning of F1, 10 animals/sex were selected from as many litters as possible to provide the F2 generation. The F3 generation was similarly derived from F2 animals. All rats were fed the same diet as their parents throughout the study. The study was terminated 2 years after the first exposure of the P generation (i.e. the F1, F2 and F3 generations were terminated at 18, 12 and 6 months).
No treatment-related adverse effects were observed on the fertility, gestation or lactation index, or on viability of the offspring in any of the three generations of offspring. No treatment-related deaths, or changes in body weight gain or food consumption were observed across the generations. There were no treatment-related differences in the levels of haemoglobin, haematocrit, blood sugar and non-protein nitrogen, urinary albumin and sugar, red and white blood cell counts, differential white cell count or prothrombin times among any of the generations. Gross pathology and microscopic examination of 15 organs and tissues, including the gonads, at study termination was unremarkable. Therefore, the NOAEL was considered to be 250 mg calcium disodium EDTA/kg bw/day, the highest tested dose.
In conclusion, calcium disodium EDTA showed no evidence of reproductive or lactational effects when fed to rats at up to 250 mg/kg bw/day (highest tested dose) in a multigeneration study. Based on the structural similarity of the two ethylenediamines, it is expected that trisodium EDDS is also unlikely to induce reproductive toxicological effects under similar test conditions.
Referenceopen allclose all
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 700 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Additional information
No information is available on fertility effects in humans.
In a GLP study conducted according to OECD Guideline 415, trisodium EDDS was assessed for its ability to induce reproductive and developmental toxicity in rats. Groups of 25 Spague-Dawley rats of each sex were given 0, 90, 250 or 700 mg/kg bw/day of the test substance (in water) by oral gavage for 70 days before mating, and throughout mating, pregnancy and weaning. Culling of the offspring occurred on day 4 after birth to standardize the litters, and the remaining offspring were killed on day 21 after delivery. The animals were observed for mortality, clinical signs of toxicity, body weight gain, feed consumption, changes in the estrus cycle, precoital index, mating index, fertility, gestation index, number and sex of offspring, litter size and viability, and lactation index. At sacrifice, gross necropsy of the thoracic, abdominal and pelvic viscera was carried out, the reproductive organs, brain and pituitary were weighed and sperm evaluations were performed. The females were examined for the number and distribution of implantation sites. Necropsies were performed on the pups culled at 4 days for external and internal abnormalities and the brains were sectioned to detect hydrocephaly. No significant systemic adverse effects were observed on the parental animals, and the reproductive and developmental parameters examined were unremarkable. Therefore the study NOAEL was considered to be 700 mg/kg bw/day (York, 1999).
No 2 -, 3- or multi-generation studies on trisodium EDDS, EDDS (or its other simple salts), or EDTA were identified.
However, In a good-quality early study (prior to GLP), calcium disodium EDTA dihydrate was assessed for the potential to cause adverse effects in a multigeneration reproductive toxicity study in Wistar rats. Groups of 25 rats of each sex (P generation) were fed 0, 50, 125 or 250 mg/kg bw/day in the diet from weaning (day 21) for up to 2 years. At 13 and 21 weeks of age, male rats were mated with females to provide two litters (F1 generation). At weaning of F1, 10 animals/sex were selected from as many litters as possible to provide the F2 generation. The F3 generation was similarly derived from F2 animals. All rats were fed the same diet as their parents throughout the study. The study was terminated 2 years after the first exposure of the P generation (i.e. the F1, F2 and F3 generations were terminated at 18, 12 and 6 months). No treatment-related adverse effects were observed on the fertility, gestation or lactation index, or on viability of the offspring in any of the three generations of offspring. No treatment-related deaths, or changes in body weight gain or food consumption were observed across the generations. There were no treatment-related differences in the levels of haemoglobin, haematocrit, blood sugar and non-protein nitrogen, urinary albumin and sugar, red and white blood cell counts, differential white cell count or prothrombin times among any of the generations. Gross pathology and microscopic examination of 15 organs and tissues, including the gonads, at study termination was unremarkable. Therefore, the NOAEL was considered to be 250 mg calcium disodium EDTA/kg bw/day, the highest tested dose. Based on the structural similarity of the two ethylenediamines, it is expected that trisodium EDDS is also unlikely to induce reproductive toxicological effects at similar dose levels (Oser et al. 1963).
Groups of Wistar rats (2 males and 4 females/group) were maintained on diets containing 0, 0.5, 1.0 or 5.0% disodium EDTA (approximately 0, 250, 500 or 2500 mg/kg bw/day, respectively) and allowed to mate upon reaching 100 days old. In order to obtain second litters, mating was repeated 10 days after weaning of the first litters. Normal first and second litters were reported from rats administered up to 500 mg/kg bw/day, while those animals of the highest dose failed to produce any litters, even after 2 months of mating. No more details were given in the brief summary, and data on the second generation were not available. Therefore, in this study, the NOAEL for reproductive effects was 500 mg/kg bw/day of disodium EDTA (Yang, 1952).
Additional information related to fertility can be obtained from a further study (Muralidhara and Narasimhamurthy, 1991). Oral administration of 5, 10 or 15 mg disodium EDTA/kg bw to male adult Swiss albino mice for five consecutive days had no effect on absolute or relative weights of epididymides and testes nor histoarchitecture of these two organs assayed at 1, 3, 5 and 7 weeks after treatment. Likewise, no effects were detected on caudal sperm counts, and there were no changes in the incidence of sperm head abnormalities or in the percentage of
abnormal sperms. Furthermore, treatment of male mice with 10 mg disodium EDTA/kg bw for five consecutive days induced no increase in the incidence of postimplantation embryonic deaths over a mating period of 8 weeks, except for a statistically insignificant about two-fold increase during week 2 and 3 of mating.
Further insight into the potential of trisodium EDDS to effect the reproductive system can be obtained from a 90-day dietary study (see IUCLID Chapter 7.5.1 for further details). In this GLP study, trisodium EDDS was fed to groups of rats (10/sex/dose) in the diet at about 0, 50, 300, 700 or 1000 mg/kg bw/day; further groups of 10 animals/sex were given the control or high-dose diets for 90 days, followed by a 28-day recovery period. The duration of the estrus cycle was determined in females over the last month of the study and sperm concentration, motility and morphology in males were also examined. At study termination, animals were examined both macroscopically and microscopically (including the epididymides, ovaries, uterus, vagina, seminal vesicles and testes), and absolute and relative organ weights determined (including the epididymides, ovaries, uterus, seminal vesicles and testes). No treatment-related effects on organ weights or length of estrus cycle were seen. In males at the top dose only, an increase in the numbers of abnormal sperm was reported (decrease in normal sperm morphololgy with a corresponding increase in head-only sperm), but by the end of the recovery period sperm morphology had returned to control levels in all but one male. No adverse effects on sperm concentration or motility were reported. Atypical residual bodies of minimal severity and incidence were noted in in the testes of the high-dose males (5/20), but these were not apparent following the 4-week recovery period. A NOAEL of 1000 mg/kg bw/day was established for trisodium EDDS based on a lack of (non-reversible) adverse effects seen on the reproductive system of rats in a 90 -day dietary study (Dotti et al. 1995).
References (not included elsewhere in IUCLID dossier - need to move to reference list in CSR)
BIBRA (1964). Summaries of toxicological data. Toxicology of EDTA. Food and Cosmetics Toxicology 2, 763-7.
EU (2004a). European Union Risk Assessment Report (RAR); edetic acid (EDTA). Vol. 49. European Chemicals Bureau (ECB). Final report available at http://ecb.jrc.ec.europa.eu/DOCUMENTS/Existing-Chemicals/RISK_ASSESSMENT/REPORT/edtareport061.pdf.
EU (2004b). European Union Risk Assessment Report (RAR); tetrasodium ethylenediaminetetraacetate (Na4EDTA). Vol. 51. European Chemicals Bureau (ECB). Final report available at http://ecb.jrc.ec.europa.eu/DOCUMENTS/Existing-Chemicals/RISK_ASSESSMENT/REPORT/na4edtareport062.pdf
Muralidhara A and Narasimhamurthy K (1991). Assessment of in vivo mutagenic potency of ethylenediaminetetraacetic acid in albino mice. Food and Cosmetics Toxicology 29, 845-849 (cited in EU, 2004a,b).
Yang S-S (1952). Toxicological investigation ofethylenediaminetetraacetic acid in the rat. Thesis dated May 1952 (cited in BIBRA, 1964).
Effects on developmental toxicity
Description of key information
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), a NOAEL of 551 mg/kg bw/day was derived for both maternal and developmental toxicity in rats given trisodium EDDS in the diet from day 6 to 15 of pregnancy (Denny, 1996a).
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), NOAELs of 400 and 1000 mg/kg bw/day were derived for maternal and developmental toxicity, respectively, in groups of rats administered trisodium EDDS by oral gavage from day 6 to 15 of pregnancy (Denny, 1996b).
Link to relevant study records
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 17 February to 12 March 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- treated to gestation day 15, not up to 1 day before birth
- GLP compliance:
- yes
- Limit test:
- no
- Species:
- rat
- Strain:
- other: Sprague-Dawley derived Crl:CD VAF/Plus
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Portage, Michigan, USA
- Age at study initiation: 12.5 weeks
- Weight at study initiation: 229-302 g
- Fasting period before study: no data
- Housing: individually (except when mating) in stainless steel, wire-mesh cages
- Diet (e.g. ad libitum): conventional, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 10 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23-25
- Humidity (%): 40-45
- Air changes (per hr): "environmentally controlled"
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 17 February To: 12 March 1994 - Route of administration:
- oral: feed
- Vehicle:
- unchanged (no vehicle)
- Details on exposure:
- DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): powdered diet (mixed with an aliquot of diet which was then added to the bulk of diet and mixed)
- Storage temperature of food: no data
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- No data in report, samples sent to sponsor for analysis
- Details on mating procedure:
- - Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: no data
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no - Duration of treatment / exposure:
- gestation day 6-15
- Frequency of treatment:
- ad libitum (in diet)
- Duration of test:
- gestation day 0-20
- No. of animals per sex per dose:
- 34 females per group (4 animals/group were killed on gestation day 16 and blood sampled for levels of copper, zinc and iron)
- Control animals:
- yes, plain diet
- Details on study design:
- - Dose selection rationale: based on a previous range-finding study
- Rationale for animal assignment (if not random): the order in which the animals were assigned corresponded to the day the copulatory plug was observed and the order of appearance on the breeding record. First mated female was assigned to the first group, the second mated female to the second group, and all the remaining animals were assigned in this manner until the required number of mated females had been placed into each group. - Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily throughout study; daily on gestational days 6 to 20.
- Cage side observations: mortality and signs of overt toxicity
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily
BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 10, 12, 16 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
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus in detail (see below); thoracic and abdominal cavities and organs subjected to gross examination for morphological changes
OTHER: four rats in each group were killed on gestation day 16 and blood samples were analysed for zinc, copper and iron. - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: foetal weights - Fetal examinations:
- - External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No - Statistics:
- Analysis of variance, Bartlett's test for homogeneity of variance and t-tests to determine the significance of differences in body weights, food consumption, mean numbers of corpora lutea, total implantations, live foetuses, gravid uterine weight and mean foetal body weights. Male to female sex ratios and proportions of litters with malformations and developmental variations were compared using the Chi-square or Fisher's test for homogeneity of R x C contingency tables to determine the significance of differences. The proportions of resorbed and dead foetuses and postimplantational losses were compared using the Kruskal-Wallis test.
- Indices:
- - Pregnancy index = No. females pregnant/ No. females mating
- Preimplantation loss = (No. corpora lutea ¿ No. implantations)/No. corpora lutea
- Postimplantation loss = (No. implantations ¿ No. viable foetuses)/ No. implantations - Historical control data:
- Historical control data is tabulated as an appendix to the report
- Details on maternal toxic effects:
- No maternal deaths occurred and no treatment related gross changes were observed. A statistically significant reduction in food consumption and body weight gain was noted for the high-dose group, as compared with the control group, during the entire dosing period, gestation days 6-15. A dose-dependent reduction in blood zinc levels was seen (which reached statistical significance at 551 mg/kg bw/day and above). Iron and copper levels were decreased in the high-dose groups, but this did not reach statistical significance. Mean gravid uterine weight for the high-dose group was significantly reduced.
There was a statistically significant increase in postimplantational losses in the high-dose group, which selectively reduced the numbers of live male foetuses, thus affecting the sex ratio. The number of early resorptions was statistically significantly higher in the high-dose group. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- ca. 551 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- body weight and weight gain
- pre and post implantation loss
- other: maternal toxicity
- Details on embryotoxic / teratogenic effects:
- Foetal body weights were significantly reduced in the high-dose group compared to the control group.
Gross malformations
A statistically significant increase in the following malformations was observed in the high dose group when compared with the incidence in the control group, (foetal incidence % control: foetal incidence % high-dose group): cleft palate (0:63), adactyly (0:33), microphthalmia (0.2:22), short lower jaw (0:21), microcephaly (0:3), anal atresia (0.2:8), malformed anal opening (0:4), omphalocele (0:3), gastroschisis (0:2.5), malformed genital tubercle (0:1.5), ectrodactyly (0:33), syndactyly (0:19), micromelia (0:11), short tail (0:5.3), bent tail (0:43), and short, bent tail (0:30). Some additional malformations were observed in the high-dose group at an incidence level above that in the control group but were not statistically significant. The numbers and type of malformations observed in the low- and mid-dose groups were similar to control levels.
In addition, subcutaneous white discolouration in the dorsal cervical and thoracic regions was observed only in the high-dose group (3.5% incidence, compared to none in the controls).
Soft tissue malformations:
Visceral examination revealed a statistically significant increase in the incidence of the following malformations in the high dose group as compared with the control group (foetal incidence % control: foetal incidence % high-dose group): short lower jaw (0:16), cleft palate (0:10), confirmed cleft palate (0:66), anophthalmia (0.4:5.2), confirmed microphthalmia (0:14), folded retina (0:14), malformed lens (0:7), internal hydrocephaly (0.4:21), malformed brain (0:3), syringomyelia (0:4), pulmonary hypoplasia and aplasia (both 0:82), malformed stomach (0:54), renal hypoplasia, malformed kidney (0:4), omphalocele (0:4), malformed intestine (0:5), malformed testes (0:7), malformed epididymis (0:5), malformed vas deferens (0:5), malpositioned testis (0:11), malformed ovary (0:17) and cavitation of the urinary and reproductive tract (0:4). Some additional visceral malformations were observed in the high-dose group at a frequency greater than the control group but were not statistically significant.
Skeletal malformations:
The incidence of several skeletal malformations occurred at a statistically significantly increased level for the high-dose group as compared with the control group, and consisted of the following (foetal incidence % control: foetal incidence % high-dose group): subcutaneous calcification (0:5), malformed skull bones (0:24), cleft palate (0:63), mandibular micrognathia (0:17), fused nasal bones (0:13), tail malformation (0.4:87), vertebral and associated rib malformation (0.4:34), bent scapula (0:66), bent clavicles (0:15), fused sternebrae (0:57), shortened rib (0:31), malformed ribs (0:5), and increased incidence of bent/malformed/absent limb bones and digits.
Additional skeletal malformations were observed in the high-dose group, but were not statistically significant.
There were instances of malformed bones observed in foetuses from the control, low-and mid-dose groups, however, the incidence level for each occurrence was one to two foetuses and no dose related increase was noted and hence these were of doubtful toxicological significance. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- ca. 551 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- reduction in number of live offspring
- changes in sex ratio
- fetal/pup body weight changes
- external malformations
- skeletal malformations
- Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Conclusions:
- In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), a NOAEL of 551 mg/kg bw/day was derived for both maternal and developmental toxicity in rats given trisodium EDDS in the diet from day 6 to 15 of pregnancy.
- Executive summary:
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), trisodium EDDS was assessed for its potential to cause maternal and developmental toxicity in Charles River rats.
Groups of 34 pregnant female rats were given trisodium EDDS in the diet at 0, 2000, 8000 or 16000 ppm (about 0, 132, 551 or 944 mg/kg bw/day based on food consumption) from gestation days 6 to 15. Four rats in each group were killed on gestation day 16 and blood samples were analysed for zinc, copper and iron. The remaining animals were killed on gestation day 20 and the dams examined for gross abnormalities and the uterus was examined for viable foetuses, early and late resorptions, number of implantations and corpora lutea. Foetuses were weighed and examined for external malformations and variations. About one-half of the foetuses were examined for soft-tissue effects and the remaining foetuses were examined for skeletal alterations.
Evidence of maternal toxicity in the high-dose group was seen as reduced body weight gain and food consumption. A dose-dependent reduction in blood zinc levels was seen (which reached statistical significance at 551 mg/kg bw/day and above). Iron and copper levels were decreased in the high-dose groups, but this did not reach statistical significance. Mean gravid uterine weight for the high-dose group was significantly reduced. There was a statistically significant increase in postimplantational losses in the high-dose group, which selectively reduced the numbers of live male foetuses, thus affecting the sex ratio. The number of early resorptions was statistically significantly increased in the high-dose group. Foetuses in the high-dose group had a diverse range of external, visceral and/or skeletal malformations and developmental variations, affecting all the major organs and skeletal structures. The whole developmental period was therefore affected by the test substance.
In conclusion, a NOAEL of 551 mg/kg bw/day was derived for both maternal and developmental toxicity in rats given trisodium EDDS in the diet from day 6 to 15 of pregnancy.
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 14 November to 8 December 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- only dosed to gestation day 15, not up to 1 day before birth
- GLP compliance:
- yes
- Limit test:
- no
- Species:
- rat
- Strain:
- other: Sprague Dawley derived Crl:CD VAF/Plus
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Portage, Michigan, USA
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 208-272 g
- Fasting period before study: no data
- Housing: individually (except when mating) in stainless steel, wire-mesh cages
- Diet (e.g. ad libitum): conventional, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 10 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24
- Humidity (%): 46-55
- Air changes (per hr): no data (environmentally controlled room)
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 14 November 1994 To: 8 December 1994 - Route of administration:
- oral: gavage
- Vehicle:
- water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: test substance weighed into a beaker and dissolved in distilled water and poured into a measuring cylinder, more water added to the required volume. Prepared weekly
VEHICLE
- Concentration in vehicle: 5, 40 or 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw/day
- Purity: distilled water - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Samples sent away for analysis to BCO Analytical Services B.V., The Netherlands; details not included in the report
- Details on mating procedure:
- - Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: no data
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no - Duration of treatment / exposure:
- 10 days (gestation days 6-15)
- Frequency of treatment:
- once daily
- Duration of test:
- gestation days 0-20
- No. of animals per sex per dose:
- 36 females per group (6 animals/group were killed on gestation day 16 and blood sampled for levels of copper, zinc and iron)
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: based on a previous range-finding study
- Rationale for animal assignment (if not random): the order in which the animals were assigned corresponded to the day the copulatory plug was observed and the order of appearance on the breeding record. First mated female was assigned to the first group, the second mated female to the second group, and all the remaining animals were assigned in this manner until the required number of mated females had been placed into each group. - Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations: mortality and signs of overt toxicity
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily
BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 16 and 20
FOOD CONSUMPTION: Yes
- Time schedule for examinations: Gestation days 0, 6, 9, 12, 16 and 20
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 16 (satellite groups) and 20
- Organs examined: uterus, placenta, kidney, spleen, ovaries
BLOOD EXAMINATION: Yes
- Plasma analysed for iron, copper and zinc levels
- Time schedule for examinations: Gestation day 16 (in 6 satellite animals/group) or 20 - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Number and location of viable and nonviable fetuses: Yes
- Other: placenta - Fetal examinations:
- - External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No data - Statistics:
- ANOVA, Dunnett's test to determine the significance of differences in body weights, food consumption, mean numbers of corpora lutea, total implantations, live foetuses, gravid uterine weight and mean foetal body weights. Proportions of litters with malformations and developmental variations were compared using the Chi-square for homogeneity of R x C contingency tables and Fisher's test to determine the significance of differences. The proportions of resorbed and dead foetuses and postimplantational losses were compared using the Kruskal-Wallis test and Mann-Whitney U test.
- Indices:
- - Pregnancy index = No. females pregnant/ No. females mating
- Preimplantation loss = (No. corpora lutea ¿ No. implantations)/No. corpora lutea
- Postimplantation loss = (No. implantations ¿ No. viable foetuses)/ No. implantations - Historical control data:
- Historical control data is tabulated in the report
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- There was an increased incidence of soft stool and decreased defecation in the 1000 mg/kg bw/day animals compared with the control group.
- Mortality:
- no mortality observed
- Description (incidence):
- No maternal deaths occurred.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Taking into account the gravid uterine weights, the mean carcass weights were reduced in all treatment groups, reaching statistical significance (p<0.05) in the high-dose group compared to the control group.
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Significantly decreased at 1000 mg/kg bw/day group during GD 6-9 and 6-16, but not statistically significant when considered over the entire study period (GD 0-20).
- Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- Post-mortem examinations: hydronephrotic kidney was observed in all groups. However, while the number of observations was slightly higher in the treated groups (1, 2, 3, 2 cases in control, low, mid and high-dose groups, respectively) there was no dose-related increase in the incidence of this finding. Other necropsy observations occurred in single instances and were not considered related to treatment.
- Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- - Uterine examination in satellite animals: no treatment-related significant effects were seen. One animal in the 400 mg/kg bw/day group was observed with higher postimplantation losses and mean number of resorptions but these effects were not considered treatment-related as they were not dose-related.
- Caesarean section examination: no treatment-related statistically significant effects were seen compared to controls.
- Blood examination: no treatment-related statistically significant changes in plasma copper, iron or zinc levels on GD 16 or 20 were observed - Dose descriptor:
- NOAEL
- Effect level:
- ca. 400 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- body weight and weight gain
- food consumption and compound intake
- Dose descriptor:
- NOAEL
- Effect level:
- ca. 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Remarks:
- No convincing evidence of adverse treatment-related malformations seen in any test
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:yes
Details on embryotoxic / teratogenic effects:
- Gross: the malformation, microphthalmia, was observed in 5 foetuses from a single litter in the 1000 mg/kg bw/day group and in 1 foetus from the control group. As all affected foetuses occurred in one litter in the top-dose group, it is questionable whether or not this malformation was related to treatment. Other external abnormalities occurred in single instances and were not considered related to treatment.
- Soft tissue: in addition to microphthalmia observed in five foetuses from one litter in the top dose group, a malformed and malpositioned eye occurred in 1 foetus and another had bilateral anophthalmia; since these animals came from the single litter in the high-dose group it is unlikely that they were treatment related. Malformed kidney was observed in 2 foetuses from the high-dose group. Visceral malformations observed in single foetuses in the high-dose group included internal hydrocephaly, malformed ureter, epididymis and vas deferens, but it is questionable whether or not they were related to treatment. A slight, but not statistically significant, increase in misshapen ovaries was seen in the 400 mg/kg bw/day group, but this effect was not dose¿related, and was not considered related to treatment.
- Skeletal: single observations of skeletal malformations included malformed skull bones, vertebral malformations, and vertebral malformations with associated rib occurring in individual foetuses from one litter in the 1000 mg/kg bw/day group. Given that all skeletal malformations were observed from the same litter, these findings were not considered to be treatment-related.
In a few instances the incidence for a particular skeletal variation reached a level which was statistically higher than that observed in the control group. Skeletal variations in the high-dose group included 25 presacral vertebrae in 14 foetuses from 4 litters, a finding which was absent in the control or the other treatment groups. Unossified sternebrae were noted in 72 foetuses from 24 litters in the high-dose group and occurred at a statistically higher incidence as compared with the control group. An extra full rib was observed in 9 foetuses from 8 litters in the high-dose group and the litter incidence was statistically greater than that observed in the control group. The variation, 7th cervical rib, was observed in 4 and 7 foetuses from 4 and 4 litters from the 400 and 1000 mg/kg bw/day groups, respectively, in comparison to a single affected foetus from the control group. However, the incidence was not significantly different from the control group. These variations are presumably evidence of developmental delay and not permanent malformations. - Key result
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Conclusions:
- In a GLP study, conducted according to EPA Guidelines, NOAELs of 400 and 1000 mg/kg bw/day were derived for maternal and developmental toxicity, respectively, in groups of rats administered trisodium EDDS by oral gavage from day 6 to 15 of pregnancy.
- Executive summary:
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), trisodium EDDS was assessed for its potential to cause maternal and developmental toxicity in Charles River rats.
Groups of 36 pregnant female rats were given trisodium EDDS (in water) by oral gavage at 0, 50, 400 or 1000 mg/kg bw/day from gestation days 6 to 15. Six (satellite) rats in each group were killed on gestation day 16 and blood samples were analysed for zinc, copper and iron. The remaining animals were killed on gestation day 20 and the dams examined for gross abnormalities and the uterus was examined for viable foetuses, early and late resorptions, number of implantations and corpora lutea. Foetuses were weighed and examined for external malformations and variations. About one-half of the foetuses were examined for soft-tissue effects and the remaining foetuses were examined for skeletal alterations.
Evidence of maternal toxicity in the high-dose group was seen as reduced carcass weight, a decrease in food consumption during the treatment period, an increased incidence of soft stools and decreased defecation. There was no effect on blood copper, zinc or iron levels. No differences in mean implantation sites, mean number of live foetuses, mean postimplantation losses or mean foetal body weights were observed between the treated and control groups.
The incidence of a particular skeletal variation in the high-dose group was statistically higher than that of the control group. Skeletal variations in this group included 25 presacral vertebrae in 14 foetuses from 4 litters, a finding which was absent in the control or the other treatment groups. Unossified sternebrae were noted in 72 foetuses from 24 litters in the high-dose group and an extra full rib was observed in 9 foetuses from 8 litters. The variation, 7th cervical rib, was observed in 4 and 7 foetuses in 4 litters each from the 400 and 1000 mg/kg bw/day groups, respectively, in comparison to a single affected foetus from the control group, however the incidence was not statistically significant. These variations were interpreted as evidence of developmental delay and not as permanent malformations.
In conclusion, a NOAEL of 400 mg/kg bw/day was derived for maternal toxicity, and a NOAEL of 1000 mg/kg bw/day for developmental toxicity, in rats given trisodium EDDS by oral gavage from day 6 to 15 of pregnancy.
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 31 August to 22 September 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- dosed days 6-15, not dosed up to oneday before study termination. Only 6 animals per group, instead of the indicated 20
- GLP compliance:
- yes
- Limit test:
- no
- Species:
- rat
- Strain:
- other: Sprague-Dawley-derived Crl:CD VAF/Plus
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Portage, Michigan
- Age at study initiation: 16 weeks
- Weight at study initiation: 236-288 g
- Fasting period before study: no
- Housing: individually, in stainless steel wire-mesh cages during experimental phase
- Diet (e.g. ad libitum): conventional, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 15 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5-22.8
- Humidity (%): 50-68
- Air changes (per hr): "regulated air changes"
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 31 August 1994 To: 22 September 1994 - Route of administration:
- oral: gavage
- Vehicle:
- water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: prepared weekly in distilled water using a magnetic stirrer.
VEHICLE
- Concentration in vehicle: 5, 20, 40, 60 or 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw/day - Analytical verification of doses or concentrations:
- not specified
- Details on analytical verification of doses or concentrations:
- none
- Details on mating procedure:
- - Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: no data
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no - Duration of treatment / exposure:
- gestation day 6-15
- Frequency of treatment:
- once daily
- Duration of test:
- gestation days 0-20
- No. of animals per sex per dose:
- 9 females/dose (3 animals from each group killed on day 16 for analysis of copper, iron and zinc levels in plasma)
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: range-finding study
- Rationale for animal assignment (if not random): random - Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 16 and 20
FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: uterus, gross examinations of organs
OTHER: blood samples were taken on day 16 for the satellite groups and on day 20 for the main groups for the analysis of copper, iron and zinc levels - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: placenta examined grossly - Fetal examinations:
- - External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: half per litter retained for possible future examination
- Head examinations: no data - Details on maternal toxic effects:
- Maternal toxic effects:no effects
Details on maternal toxic effects:
No animals died during the study and no treatment-related clinical signs of toxicity were observed. At necropsy individual findings were not considered to be related to ttreatment. There were no significant differences in mean body weights or body weight gain between the groups either overall or during the dosing period and food consumption was comparable between groups. Plasma copper, iron and zinc levels were similar in the treated and control group.
Pregnancy rates were similar in all of the main groups; in the satellite groups treated at 50 or 200 mg/kg bw/day, one and two animals respectively were not pregnant. Postplantational losses were highest in the control group and no late resorptions or dead foetuses were observed in any of the groups. Gravid uterine weights were comparable for all groups. - Dose descriptor:
- NOAEL
- Effect level:
- ca. 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
No treatment-related findings were evident on external examination of the foetuses or on examination of the soft tissues at any dose. - Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Conclusions:
- In a GLP range-finding study, conducted to a protocol similar to OECD Guideline 414, trisodium EDDS showed no evidence of maternal or developmental toxicity when administered by oral gavage to pregnant rats on gestation days 6-15 at up to 1000 mg/kg bw/day, considered the study NOAEL.
- Executive summary:
In a GLP range-finding study, conducted to a protocol similar to OECD Guideline 414, the ability of trisodium EDDS to cause maternal and developmental toxicity was assessed in a oral range-finding study in pregnant Sprague-Dawley-derived rats.
Groups of 9 females were given 0, 50, 200, 400, 600 or 1000 mg/kg bw/day by oral gavage on gestation days 6-15. Three animals per group were killed on gestation day 16 and blood samples taken for the analysis of plasma copper, iron and zinc levels. The remaining animals were killed on gestation day 20, blood samples taken for mineral levels, and the thoracic and abdominal organs were grossly examined for abnormalities. The uteri were removed and examined for the number of viable foetuses, early and late resorptions, total implantations and corpora lutea. The gravid uterine weights were recorded. Foetuses were weighed, sexed and examined for external abnormalities and in half of the foetuses for soft tissue changes. The dams were weighed at intervals throughout the study and food consumption recorded.
No animals died during the study and no treatment-related clinical signs of toxicity were observed. At necropsy, individual findings were not considered to be treatment-related. There were no significant differences in mean body weights or body weight gain between the groups either overall or during the dosing period and food consumption was comparable between groups. Plasma copper, iron and zinc levels were similar in the treated and control group. Pregnancy rates were similar in all of the main groups; in the satellite groups treated at 50 or 200 mg/kg bw/day, one and two animals respectively were not pregnant. Postplantational losses were highest in the control group and no late resorptions or dead foetuses were observed in any of the groups. Gravid uterine weights were comparable for all groups. No treatment-related findings were evident on external examination of the foetuses or on examination of the soft tissues.
In conclusion, trisodium EDDS showed no evidence of maternal or developmental toxicity when administered by oral gavage to pregnant rats on gestation days 6-15 at up to 1000 mg/kg bw/day, considered the study NOAEL.
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 17 January to 3 February 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- only 5 animals per dose (instead of 20); dosed until day 15, not to one day before sacrifice
- GLP compliance:
- yes
- Limit test:
- no
- Species:
- rat
- Strain:
- other: Sprague-Dawley-derived Crl:CD VAF/Plus
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Portage, Michigan
- Age at study initiation: 12.5 weeks
- Weight at study initiation: 232-264 g
- Fasting period before study: no
- Housing: individually, in stainless steel wire-mesh cages during experimental phase
- Diet (e.g. ad libitum): conventional, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 10 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-23
- Humidity (%): 34-66
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 17 January 2010 To: 17 January 3 February 2010 - Route of administration:
- oral: feed
- Vehicle:
- unchanged (no vehicle)
- Analytical verification of doses or concentrations:
- yes
- Details on mating procedure:
- - Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: no data
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no - Duration of treatment / exposure:
- gestation days 6-15
- Frequency of treatment:
- daily
- Duration of test:
- 20 days
- No. of animals per sex per dose:
- 5 females per dose
- Control animals:
- yes, plain diet
- Details on study design:
- - Dose selection rationale: selected by the sponsor based on data available from other studies
- Rationale for animal assignment (if not random): random - Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily
BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 16 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: no data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: uterus removed for examination; thoracic and abdominal organs grossly examined
- Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: uterine content examined
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes - Fetal examinations:
- None
- Details on maternal toxic effects:
- Two deaths occurred in the 40000 ppm dose group. There was a considerable decrease in food consumption in the 24000 and 40000 ppm groups during the treatment period (days 6-15) and about a 20% reduction at 16000 ppm compared to the untreated period (see Table 1). Mean body weight gain by day 16 was 42 g at 16000 ppm compared to 70 g in the control group and mean body weight loss was 35 and 64 g at 24000 and 40000 ppm, respectively. One and five animals were emaciated at 24000 and 40000 ppm, respectively; soft feces and reduced defecation also occurred in these groups and one animal at 40000 ppm produced no feces at all. Body surface staining was seen in one and three animals at 8000 and 40000 ppm, respectively. There were no treatment-related observations at necropsy.
No viable fetuses were observed at the highest dose level. The mean number of live fetuses per animal was reduced to five at 24000 ppm compared to 14.5 in the control group. The mean number of live fetuses was similar at the lower doses to the control group. Gravid uterine weights were reduced at 16000 and 24000 ppm. The mean number of total resorptions was 0.5, 0.2, 1.6, 0.8 and 7.67 at 0, 2000, 8000, 16000 and 24000 ppm, respectively. Late resorptions were only observed in the 24000 and 40000 ppm groups. At 24000 ppm postimplantational losses were about 14-fold higher than in the controls. - Dose descriptor:
- NOAEL
- Effect level:
- ca. 8 000 ppm (nominal)
- Based on:
- test mat.
- Basis for effect level:
- other: maternal toxicity
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:yes
Details on embryotoxic / teratogenic effects:
Postplantational losses were 9.4, 5.4, 59.8 and 100% at 8000, 16000, 24000 and 40000 ppm respectively, compared to 3.6% in the control group. However, according to the report (due to the reduction in food consumption at the higher doses) the test substance intake levels were 530, 981, 830 and 1029 mg/kg bw, respectively for these groups. Thus, the increase in postimplantational losses may have been due to the considerably reduced food intake (and corresponding mean body weight lossess) at the two highest doses. Therefore, a NOAEL could not be determined for developmental toxicity. - Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Conclusions:
- In a GLP range-finding study, conducted to a protocol similar to OECD Guideline 414, trisodium EDDS given in the diet to pregnant female rats at 0, 2000, 8000, 16000, 24000 or 40000 ppm resulted in maternal toxicity at 16000 ppm and above. Therefore, a NOAEL for maternal toxicity of 8000 ppm, about 530 mg/kg bw/day, was determined. Due to the considerable reduction of food consumption at 24000 and 40000 ppm (and corresponding body weight losses at these high doses), developmental toxicity could not accurately be assessed, but did not occurr at non-maternally toxic levels.
- Executive summary:
In a GLP study, conducted to a protocol similar to OECD Guideline 414, trisodium EDDS was assessed for maternal and development toxicity in a range-finding, dietary study in pregnant Charles River rats.
The dams were given 0, 2000, 8000, 16000, 24000 or 40000 ppm of the test material in the diet from days 6-15 of pregnancy. The animals were observed daily for clinical signs of toxicity and body weights and food consumption were recorded at intervals throughout the study. On day 20 the animals were killed and uterine examinations carried out for the number of viable foetuses, early and late resorptions and the number of total implantations and corpora lutea. Gravid uterine weights were recorded. The organs of the abdominal and thoracic cavities were examined for gross abnormalities.
Two deaths occurred in the top dose group. There was a considerable decrease in food consumption in the 24000 and 40000 ppm groups during the treatment period (days 6-15) and about a 20% reduction at 16000 ppm compared to consumption during the untreated period. Mean body weight gain on day 16 was reduced at 16000 ppm and mean body weight losses occurred at 24000 and 40000 ppm. One and five animals were emaciated at 24000 and 40000 ppm, respectively, soft feces and reduced defecation also occurred in these groups and one animal at 40000 ppm produced no feces at all. No viable foetuses were observed at the highest dose level. The mean number of live fetuses per animal was reduced to five at 24000 ppm compared to 14.5 in the control group, with those at the lower doses being similar to the controls. Gravid uterine weights were reduced at 16000 and 24000 ppm. The mean number of total resorptions was 0.5, 0.2, 1.6, 0.8 and 7.67 at 0, 2000, 8000, 16000 and 24000 ppm, respectively. Late resorptions were only observed in the 24000 and 40000 ppm groups. At 24000 ppm postimplantational losses were about 14-fold higher than in the controls.
In conclusion, trisodium EDDS given in the diet to pregnant female rats at up to 40000 ppm resulted in maternal toxicity at 16000 ppm and above. Therefore, a NOAEL for maternal toxicity of 8000 ppm, about 530 mg/kg bw/day, was determined in this range-finding study. Due to the considerable reduction of food consumption at 24000 and 40000 ppm (and corresponding body weight lossess at these high doses), developmental toxicity could not accurately be assessed.
- Endpoint:
- developmental toxicity
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- the study does not need to be conducted because the substance is of low toxicological activity (no evidence of toxicity seen in any of the tests available), it can be proven from toxicokinetic data that no systemic absorption occurs via relevant routes of exposure (e.g. plasma/blood concentrations below detection limit using a sensitive method and absence of the substance and of metabolites of the substance in urine, bile or exhaled air) and there is no or no significant human exposure
- Species:
- rabbit
Referenceopen allclose all
Table 1. Mean dam food consumption (g/animal/day)
| Days | 0 ppm | 2000 ppm | 8000 ppm | 16000 ppm | 24000 ppm | 40000 ppm |
| 0-6 | 19.1 ± 2.0 | 18.1 ± 1.0 | 18.1 ± 1.5 | 19.9 ± 2.8 | 19.6 ± 1.3 | 18.5 ± 3.9 |
| 6-9 | 21.4 ± 2.3 | 20.5 ± 0.5 | 19.7 ± 1.4 | 17.3 ± 0.9 | 8.6 ± 0.6 | 6.6 ± 0.9 |
| 9-12 | 21.4 ± 1.9 | 21.4 ± 1.5 | 20.5 ± 1.8 | 17.5 ± 1.9 | 6.6 ± 2.7 | 2.0 ± 1.2 |
| 12-16 | 22.8 ± 1.3 | 22.0 ± 1.4 | 22.1 ± 1.9 | 18.3 ± 1.7 | 6.9 ± 5.1 | 5.5 ± 0.6 |
| 16-20 | 24.8 ± 2.1 | 23.5 ± 1.0 | 25.5 ± 2.2 | 23.5 ± 3.9 | 26.5 ± 4.4 | 22.1 ± 2.2 |
| 6-16 | 21.9 ± 1.6 | 21.3 ± 1.0 | 20.8 ± 1.5 | 17.7 ± 1.4 | 7.3 ± 2.5 | 4.3 ± 1.2 |
| 0-20 | 22.3 ± 1.6 | 21.4 ± 1.1 | 21.5 ± 1.4 | 19.3 ± 1.2 | 12.9 ± 2.7 | 9.2 ± 2.4 |
Uncertainties resulting from reduced food consumption at the higher dose levels made it difficult to determine which effects were due to treatment-related toxicity and which were due to poor nutrition
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 551 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Additional information
No information is available on developmental effects in humans.
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), trisodium EDDS was assessed for its potential to cause maternal and developmental toxicity in Charles River rats. Groups of 34 pregnant female rats were given trisodium EDDS in the diet at 0, 2000, 8000 or 16000 ppm (about 0, 132, 551 or 944 mg/kg bw/day based on food consumption) from gestation days 6 to 15. Four rats in each group were killed on gestation day 16 and blood samples were analysed for zinc, copper and iron. The remaining animals were killed on gestation day 20 and the dams examined for gross abnormalities and the uterus was examined for viable foetuses, early and late resorptions, number of implantations and corpora lutea. Foetuses were weighed and examined for external malformations and variations. About one-half of the foetuses were examined for soft-tissue effects and the remaining foetuses were examined for skeletal alterations. Evidence of maternal toxicity in the high-dose group was seen as reduced body weight gain and food consumption. A dose-dependent reduction in blood zinc levels was seen (which reached statistical significance at 551 mg/kg bw/day and above). Iron and copper levels were decreased in the high-dose groups, but this did not reach statistical significance. Mean gravid uterine weight for the high-dose group was significantly reduced. There was a statistically significant increase in postimplantational losses in the high-dose group, which selectively reduced the numbers of live male foetuses, thus affecting the sex ratio. The number of early resorptions was statistically significantly increased in the high-dose group. Foetuses in the high-dose group had a diverse range of external, visceral and/or skeletal malformations and developmental variations, affecting all the major organs and skeletal structures. The whole developmental period was therefore affected by the test substance. In conclusion, a NOAEL of 551 mg/kg bw/day was derived for both maternal and developmental toxicity in rats given trisodium EDDS in the diet from day 6 to 15 of pregnancy (Denny, 1996a).
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), trisodium EDDS was assessed for its potential to cause maternal and developmental toxicity in Charles River rats. Groups of 36 pregnant female rats were given trisodium EDDS (in water) by oral gavage at 0, 50, 400 or 1000 mg/kg bw/day from gestation days 6 to 15. Six (satellite) rats in each group were killed on gestation day 16 and blood samples were analysed for zinc, copper and iron. The remaining animals were killed on gestation day 20 and the dams examined for gross abnormalities and the uterus was examined for viable foetuses, early and late resorptions, number of implantations and corpora lutea. Foetuses were weighed and examined for external malformations and variations. About one-half of the foetuses were examined for soft-tissue effects and the remaining foetuses were examined for skeletal alterations. Evidence of maternal toxicity in the high-dose group was seen as reduced carcass weight, a decrease in food consumption during the treatment period, an increased incidence of soft stools and decreased defecation. There was no effect on blood copper, zinc or iron levels. No differences in mean implantation sites, mean number of live foetuses, mean postimplantation losses or mean foetal body weights were observed between the treated and control groups. The incidence of a particular skeletal variation in the high-dose group was statistically higher than that of the control group. Skeletal variations in this group included 25 presacral vertebrae in 14 foetuses from 4 litters, a finding which was absent in the control or the other treatment groups. Unossified sternebrae were noted in 72 foetuses from 24 litters in the high-dose group and an extra full rib was observed in 9 foetuses from 8 litters. The variation, 7th cervical rib, was observed in 4 and 7 foetuses in 4 litters each from the 400 and 1000 mg/kg bw/day groups, respectively, in comparison to a single affected foetus from the control group, however the incidence was not statistically significant. These variations were interpreted as evidence of developmental delay and not as permanent malformations. In conclusion, a NOAEL of 400 mg/kg bw/day was derived for maternal toxicity, and a NOAEL of 1000 mg/kg bw/day for developmental toxicity, in rats given trisodium EDDS by oral gavage from day 6 to 15 of pregnancy (Denny, 1996b).
In a GLP range-finding study, conducted to a protocol similar to OECD Guideline 414, trisodium EDDS given in the diet to pregnant female rats at 0, 2000, 8000, 16000, 24000 or 40000 ppm resulted in maternal toxicity at 16000 ppm and above. Therefore, a NOAEL for maternal toxicity of 8000 ppm, about 530 mg/kg bw/day, was determined. Due to the considerable reduction of food consumption at 24000 and 40000 ppm (and corresponding body weight losses at these high doses), developmental toxicity could not accurately be assessed, but did not occurr at non-maternally toxic levels (Denny, 1996c).
In a GLP range-finding study, conducted to a protocol similar to OECD Guideline 414, trisodium EDDS showed no evidence of maternal or developmental toxicity when administered by oral gavage to pregnant rats on gestation days 6-15 at up to 1000 mg/kg bw/day, considered the study NOAEL (Denny, 1996d).
Toxicity to reproduction: other studies
Additional information
In a GLP study, conducted according to EPA Guidelines (OPPTS 870.3700), trisodium EDDS was assessed for its potential to cause maternal toxicity and alter plasma levels of copper, iron and zinc following oral administration in pregnant Charles River rats. Groups of 15 female rats were administered 0, 50, 400 or 1000 mg/kg bw/day of the test substance by oral gavage from gestation day 6 to 15. The animals were observed for signs of clinical toxicity throughout the study and at regular intervals body weights and food consumption were recorded. Blood samples were taken 2 and 4 h after the last dose on day 15 and analysed for plasma levels of copper, iron and zinc. The dams were examined for gross abnormalities of the major organs; no examination of the fetuses took place. No deaths occurred during the study. Soft feces were observed in five animals at the high-dose but no other clinical observations were considered to be treatment-related. Body weight gain was slightly reduced in a dose-dependent manner and food consumption was reduced for the first three days of dosing in the high-dose group. A statistically significant, dose-related reduction in plasma zinc levels was evident in all treatment groups compared to the controls at both the 2 and 4 h sampling times. Plasma copper levels were reduced in all treatment groups, reaching significance at 4 h in the low-dose group and at both time points in the mid- and high-dose groups. No statistically significant or dose-related reduction in plasma iron levels were seen. In conclusion, trisodium EDDS caused maternal toxicity, evident as soft feces and a slight reduction in body weight gain at 1000 mg/kg bw/day, and caused a dose-related, statistically significant reduction in plasma levels of copper and zinc when given orally to pregnant rats on days 6-15 of gestation. No statistically significant or dose-related reduction in plasma iron levels were seen (Denny, 1996e).
Justification for classification or non-classification
According to EU CLP regulation, trisodium EDDS would not meet the requirement for classification as a developmental or reproductive toxicant, under the test conditions described.
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
