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Description of key information

The acute oral LD50 of edetic acid in rats was found to be 4500 mg/kg bw (BASF SE, 1973). The LOAEC established in an inhalation study with Na2EDTAwas considered to be 30 mg/m³ air. Furthermore, it is highly unlikely that EDTA induced acute dermal toxicity as neither Ca or Na salts of EDTA are able to penetrate the skin.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Principles of method if other than guideline:
BASF-TEST: In principle, the methods described in the OECD Guideline 401 were used. Several groups of 5 rats per sex and dose were treated simultaneously by gavage with preparations of the test substance in suitable vehicle. The concentrations of these preparations were used to achieve comparable volumes per kg body weight. Group-wise documentation of clinical signs was performed over the 14 day study period. Body weight was determined before the start of the study only, as it was needed for determination of dose.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
OTHER SPECIFICS:
- Name of test material (as cited in study report): Trilon BS
Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Mean body weight at study initiation: males: 264 g; females: 274 g

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
DOSAGE PREPARATION
- Stock solution prepared: 30 %
- Dose volume applied: 10.66 mL/kg bw for the 3200 mg/kg bw dose group; 13.33 mL/kg bw for the 4000 mg/kg bw dose group; 16.66 mL/kg bw for the 5000 mg/kg bw dose group; 21.4 mL/kg bw for the 6400 mg/kg bw dose group; 26.6 mL/kg bw for the 8000 mg/kg bw dose group; 33.3 mL/kg bw for the 10000 mg/kg bw dose group.
Doses:
3200; 4000; 5000; 6400; 8000; 10000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days;
- The animals were observed for mortality and clinical signs of toxicity;
- Frequency of observations: Several times on the application day, thereafter once each working day;
- Body weights were only recorded at the beginning of the study;
- Necropsy of survivors and animals which died performed: yes
Sex:
male/female
Dose descriptor:
LD50
Effect level:
4 500 mg/kg bw
Mortality:
1/10 died in the 3200 mg/kg bw dose group, 3/10 died in the 4000 mg/kg bw dose group, 6/10 died in the 5000 mg/kg bw dose group, 8/10 died in the 6400 mg/kg bw dose group, 9/10 died in the 8000 mg/kg bw dose group, 10/ 10 died in the 10000 mg/kg bw dose group (See table 1).
Clinical signs:
3200 mg/kg bw and 4000 mg/kg bw dose group: directly after application accelerated respiration, squatting posture, red eyes; 1 day later: squatting posture, aggressiveness, contaminated fur, slight diarrhea; 5 days later: aggressiveness, contaminated fur; fully reversible within 14 days.
5000 mg/kg bw and 6400 mg/kg bw dose group: directly after application accelerated respiration, squatting posture, red eyes; 1 day later: squatting posture, aggressiveness, contaminated fur, slight diarrhea; 5 days later: squatting posture, aggressiveness, contaminated fur, diarrhea, crusted muzzle and eyes; fully reversible within 13 days
8000 mg/kg bw dose group: directly after application accelerated respiration, squatting posture, red eyes, slight diarrhea, spastic gait, reduced movement; 1 day later: squatting posture, red eyes, diarrhea, contaminated fur, aggressiveness; 5 days later: aggressiveness, squatting posture, diarrhea, contaminated fur, crusted muzzle and eyes; fully reversible within 14 days in the surviving animal
10000 mg/kg bw dose group: directly after application accelerated respiration, squatting posture, red eyes, slight diarrhea, spastic gait, mortality within 90 min.
Body weight:
Body weights were not recorded during and at the end of the observation period.
Gross pathology:
Animals which died:
10000 mg/kg bw dose group: heart: acute dilatation, venous hyperemia; kidneys: muddy gray
Remaining dose groups: heart: acute dilatation particularly right, venous hyperemia; stomach: dilatated, dry hard content, bloody ulceration in the corpus; gut: diarrhea,
Animals sacrificed:
-nothing abnormal detected

Table 1: Mortalities of rats after oral application of EDTA

3200 mg/kg bw 4000 mg/kg bw 5000 mg/kg bw 6400 mg/kg bw 8000 mg/kg bw 10000 mg/kg bw
1 h male  0/5 0/5 0/5 0/5 0/5 0/5
female 0/5 0/5 0/5 0/5 0/5 0/5
24 h male  0/5 0/5 0/5 0/5 1/5 5/5
female 0/5 0/5 0/5 0/5 1/5 5/5
48 h male  0/5 0/5 0/5 1/5 1/5 5/5
female 0/5 0/5 0/5 1/5 1/5 5/5
7 d male  1/5 3/5 5/5 5/5 5/5 5/5
female 0/5 0/5 0/5 3/5 4/5 5/5
14 d male  1/5 3/5 5/5 5/5 5/5 5/5
female 0/5 0/5 1/5 3/5 4/5 5/5
Interpretation of results:
other: Classification criteria of Regulation (EC) No 1272/2008 not met
Endpoint conclusion
Dose descriptor:
LD50
Value:
4 500 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 412 (Repeated Dose Inhalation Toxicity: 28/14-Day)
Deviations:
yes
Remarks:
Dosing until day 5 only
GLP compliance:
yes (incl. QA statement)
Test type:
other: subacute
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch Number: 06088797V0

OTHER SPECIFICS
- Expiry date: 1 September 2011
- Appearance: Solid white powder
Species:
rat
Strain:
Wistar
Sex:
male
Details on test animals or test system and environmental conditions:
Age: 7 weeks (approx)
Identification: Tattooing of ears
All animals free of disease and clinical signs
Rats housed together (5 animals per cage) in Polysulfon cages
Bedding: Type Lignocel fibres, dust free bedding
Woodne gnawing blocks for enrichment
Rooms: Fully ariconditioned, temperature range 20 to 24 degrees celcius, 30 to 70% humidity
Light/dark cycle of 12 hours (6 am to 6pm light, 6 pm to 6 am dark)
Food, drinking water and bedding/enrichment materials were analysed for chemical and microbiological contaminants.
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose/head only
Vehicle:
air
Details on inhalation exposure:
A dust aerosol was generated using a dust generator and compressed air inside a mixing stage mixed with conditioned dilution air and passed into the inhalation system. The test substance was mixed with Aerosil R972 prior to facilitate aerosol generation.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Concentrations of the inhalation atmospheres were analyzed using gravimetry. Daily means were calculated based on 2 measured samples per concentration and exposure.
Duration of exposure:
6 h
Remarks on duration:
per day, 5 consecutive days
Concentrations:
- 30, 300, 1000 mg/m³ (nominal conc. of Na2H2EDTA)
- 33.3 (± 2.3), 320 (± 27), 1103 (± 52) mg/m3 (measured (with SD) referring to test substance Na2H2EDTA x 2 H2O)
No. of animals per sex per dose:
10 animals per dose group
An additional 10 animals for the high dose group and control
Control animals:
yes
Details on study design:
The animals were exposed to a respirable dust aerosol for 6 hours per day for 5 consecutive days. The exception was the high dose group (1000mg/m3) where exposure was for one day only due to mortality observed.
Statistics:
Body weight/body weight change, food consumption - comparison of each group with control using DUNNETTS test (two-sided) for the hypothesis of equal means. Clinical pathology, urine volumes, urine specific gravity, weight parameters - non-parametric one-way analysis using Kruskal-wallis test (two sided). If resulting p-value was less than 0.05, a pairwise comparison of each dose group with the control group was performed using Wilcoxon-test (two sided) for equal means.
Key result
Sex:
male
Dose descriptor:
other: LOAEC
Effect level:
ca. 30 mg/m³ air
Based on:
test mat.
Remarks on result:
other: Basis for effect level: histopathology of the respiratory tract and lung weights
Mortality:
6 deaths in the high dose group on days 0 and 1. Accelerated respiration, respiration sounds, piloerection, red encrusted nose, hunched position; Mid dose group - accelerated respiration, respiration sounds, piloerection, reduced fur care
Clinical signs:
other: 6 deaths in the high dose group on days 0 and 1. Accelerated respiration, respiration sounds, piloerection, red encrusted nose, hunched position; Mid dose group - accelerated respiration, respiration sounds, piloerection, reduced fur care
Body weight:
Decreased bodyweight change in mid and high dose group
Gross pathology:
Congestion, edema and multifocal hemorraghes in lungs of high dose group
Other findings:
FOOD CONSUMPTION
- decreased food consumption between days 0 and 1 in mid and high dose group

ORGAN WEIGHTS
- Lung weight increase in low and mid dose group

DETAILS ON RESULTS

Histopathology results:

High dose: Multifocal hemorraghes in the lungs; Inflammatory cell infiltrates

Mid dose:

Larynx: laryngeal, epithelial necrosis, multifocal, in various levels of the larynx

Inflammatory cell infiltrates in various levels of the larynx

laryngeal squamous metaplasia, multifocal, in various levels of the larynx

Regenerative hyperplasia of the laryngeal epithelium, multifocal, in various levels of the larynx

Lungs: Regenerative hyperplasia of bronchiolar epithelium (predominantly: medium bronchi, terminal bronchioles)

Mucous cell hyperplasia in large bronchi

interstitial infiltration of eosinophylic granulocytic cells

Low dose:

Larynx: Laryngeal, epithelial necrosis, multifocal, at the base of the epiglittis (level 1)

Inflammatory cell infiltrates at the base of the epiglottis (level 1)

Lungs: Regenerative hyperplasia of the bronchiolar epithelium (predominantly medium bronchi and terminal bronchioles)

Mucous cell hyperplasia in large bronchi

interstitial infiltration of eosinophylic granulocytic cells.

There were no histopathological findings in any of the recovery group animals. Thus, all pathology was reversible within the recovery period.

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Inhalation exposure to 1000 mg/m3 disodium EDTA for 6 hours caused lethality in 6 out of 20 male rats. Histological examination of the lung of the dead rats revealed congestion, edema, multifocal hemorraghes and inflammatory cell infiltrates. Inhalation exposure of rats to disodium EDTA for 6 hours per day, 5 consecutive days cause concentration dependant lesions inthe larynx and lungs that were fully reversible within 14 days. Due to histopahological changes in the low dose group a no observed effect level could not be determined.
Executive summary:

In a subacute repeated dose toxicity study (BASF, 2009) 10 male Wistar rats per dose were exposed to a respirable dust aerosol of Na2H2EDTA for 6 hours per day for 5 consecutive days at concentrations of 0, 30, 300, 1000 mg/m³ air (also see capter 7.5).

Exposure in the high dose group (1000 mg/m3) was for one day only due to mortality observed. Inhalation exposure to 1000 mg/m³ disodium EDTA for 6 hours caused lethality in 6 out of 20 male rats. Histological examination of the lung of the dead rats revealed congestion, edema, multifocal hemorraghes and inflammatory cell infiltrates.

Inhalation exposure of rats to disodium EDTA for 6 hours per day, 5 consecutive days caused concentration dependant lesions in the larynx and lungs that were fully reversible within 14 days. Due to histopahological changes in the low dose group a no observed effect level could not be determined.

The LOAEC was considered to be 30 mg/m³ air.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed

Additional information

ORAL

In the key study (BASF SE, 1973) single doses of 3200, 4000, 5000, 6400, 8000 and 10000 mg/kg bw edetic acid were administered by gavage to male and females rats as 30% solution in carboxymethyl cellulose solution. The dose groups consisted of 5 males and 5 females each and the animals were observed for 14 days. The LC50 was found to be 4500 mg/kg bw. Clinical symptoms were: squatting posture, aggressiveness, diarrhea and contaminated fur in all dose group. Autopsy of the animals which died revealed acute heart dilatation, bloody ulceration of the stomach and soft to fluid contents of the intestine.

An additional acute oral toxicity study (Akzo Chemicals, 1987) obtained a LD50 > 2000 mg/kg bw for male and female rats. No clinical signs or autopsy findings have been reported up to 2000 mg/kg bw.

INHALATION

In a subacute repeated dose toxicity study (BASF SE, 2010) 10 male Wistar rats per dose were exposed to a respirable dust aerosol of Na2H2EDTA for 6 hours per day for 5 consecutive days at concentrations of 0, 30, 300, 1000 mg/m³ air (also see chapter 7.5).

Exposure in the high dose group (1000 mg/m³) was for one day only due to mortality observed. Inhalation exposure to 1000 mg/m³ disodium EDTA for 6 hours caused lethality in 6 out of 20 male rats. Histological examination of the lung of the dead rats revealed congestion, edema, multifocal hemorraghes and inflammatory cell infiltrates.

Inhalation exposure of rats to disodium EDTA for 6 hours per day, 5 consecutive days cause concentration dependant lesions in the larynx and lungs that were fully reversible within 14 days. Due to histopahological changes in the low dose group a no observed effect level could not be determined.

The LOAEC was considered to be 30 mg/m³ air.

Justification for classification or non-classification

SELF-CLASSIFICATION ACCORDING TO REGULATION (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result, the substance is considered to be classified as Acute Toxicity Inhalation Cat. 4 (H332, harmful if inhaled) but not classified for acute oral and acute dermal toxicity under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation (EU) No 2016/1179.

CLASSIFICATION ACCORDING TO ANNEX VI OF REGULATION (EC) No 1272/2008.

The substance is not officially classified for acute oral, dermal or inhalation toxicity under Regulation (EC) No. 1272/2008, as amended for the ninth time in Regulation (EU) No 2016/1179.