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Description of key information

oral

LD50 of Na4EDTA was found to be > 1780 mg/kg bw (BASF 1983)

Inhalative 

LOAEC value of 30 mg/m³ air was established in a subacute range finder toxicity study with Na2H2EDTA, which here is also suitable to assess acute toxicity.

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. Young adult laboratory rats were purchased from breeder. Several groups of 10 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 as well as after day 2, 5, 7 and 13.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
- Lot/batch No.: 03370
- Impurities (identity and concentrations): 8% Na3 Ethylenediaminetriacetic acid, 1.2% Na2 Ethylenediaminetriacetic acid, 3.6% Na3 Nitriloltriacetic acid, < 0.3% Na2 iminodiacetic acid, 2.6% Na-glycolic acid, urotropin
- Expiration date of the lot/batch: April 1984
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Biberach, Germany
- Weight at study initiation:
1210 mg/kg bw: 179 g males/178 g females
1780 mg/kg bw: 188 g males/180 g females
2000 mg/kg bw: 180 g males/182 g females
2610 mg/kg bw: 188 g males/187g females
- Housing: 5 animals per cage
- Diet: Ssniff R, ad libitum
- Water: ad libitum
- Acclimation period: at least one week
- Fasting period before study: 16 h

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24 °C
- Humidity (%): 30 - 70%
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
DOSAGE PREPARATION
- Stock solutions prepared:
12.1% for the 1210 mg/kg bw dose group
17.8% for the 1780 mg/kg bw dose group
20.0% for the 2000 mg/kg bw dose group
26.1% for the 2610 mg/kg bw dose group

DOSE VOLUME APPLIED:
10 mL/kg bw
Doses:
1210 mg/kg bw; 1780 mg/kg bw; 2000 mg/kg bw; 2610 mg/kg bw
No. of animals per sex per dose:
10
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Frequency of weighing: prior to the start of the experiment and on day 2, 5, 7, 13
- Necropsy of survivors performed: yes
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 780 - < 2 000 mg/kg bw
Sex:
male
Dose descriptor:
LD50
Effect level:
1 913 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
1 780 mg/kg bw
Mortality:
No mortalities were observed in the lowest dose group. At 2000 mg/kg bw all females and 7/10 males died (see table 1). In the highest dose group still half of the males survived.
Clinical signs:
1210 mg/kg bw: no effects
1780 mg/kg bw: males dyspnea, apathy, ataxia, shaggy fur, poor general state; fully reversible within 5 days /females dyspnea, apathy, ataxia, abnormal positions, spastic gait, exiccosis, diarrhea, shaggy fur, saliva, poor general state; fully reversible within 1 day
2000 mg/kg bw: males dyspnea, apathy, ataxia, shaggy fur, poor general state; fully reversible within 1 day/females dyspnea, apathy, ataxia, atony, exiccosis, poor general state
2610 mg/kg bw: males dyspnea, apathy, ataxia, aggressiveness, diarrhea, piloerrection, poor general state; fully revesible within 1 day/females: dyspnea, apathy, ataxia, atony, shaggy fur, poor general state
Body weight:
1210 mg/kg bw: body weight gain male 106 g/ 46 g female
1780 mg/kg bw: body weight gain male 95 g/ 47 g female
2000 mg/kg bw: body weight gain male 93 g/ - (all animals died)
2610 mg/kg bw: body weight gain male 97 g/ - (all animals died)
Gross pathology:
Animals that died:
stomach: redness and/or bloody ulceration of the glandular part of the stomach, redness of the mucous membrane, general hyperemia
gut: atonic, redness of the mucous membrane, bloody mucous content, general hyperemia
Animals which were sacrificed:
nothing abnormal detected

Table 1: Mortalities of rats after oral application of Na4EDTA

1210 mg/kg bw 1780 mg/kg bw 2000 mg/kg bw  2610 mg/kg bw
1 h male  0/10 0/10 3/10 0/10
female 0/10 0/10 1/10 6/10
24 h male  0/10 2/10 7/10 5/10
female 0/10 4/10 10/10 10/10
48 h male  0/10 2/10 7/10 5/10
female 0/10 4/10 10/10 10/10
7 d male  0/10 2/10 7/10 5/10
female 0/10 4/10 10/10 10/10
14 d male  0/10 2/10 7/10 5/10
female 0/10 4/10 10/10 10/10
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 780 mg/kg bw
Quality of whole database:
Study similar to OECD TG 401

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:
Batch Number: 06088797V0
Expiry date: 01 September 2011
Species:
rat
Strain:
Wistar
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: 7 weeks (approx)
- Housing: housed together (5 animals per cage) in Polysulfon cages:Type Lignocel fibres, dust free bedding; Wooden gnawing blocks for enrichment

ENVIRONMENTAL CONDITIONS
Rooms: fully ariconditioned
- Temperature (°C): 20 to 24°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): cycle of 12 hours (6 am to 6pm light, 6pm to 6 am dark)

Identification: Tattooing of ears
All animals free of disease and clinical signs. 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. From the Daily mean values of each concentration, mean concentrations and standard deviati
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:
act. ingr.
Remarks:
Na2H2EDTA
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

HISTOPATHOLOGY
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.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Value:
30 mg/m³
Quality of whole database:
OECD TG 412 (Read-Across)

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Oral route

In the key study conducted by BASF AG (1983) single doses of 1210, 1780, 2000, 2610 mg/kg bw Na4EDTA were administered by gavage to male and females rats as 12 - 26% solution in water. The dose groups consisted of 10 males and 10 females each which were fasted 16 prior to the experiment. After administration a 14 day observation period followed. The LC50 was found to be between 1780 and 2000 mg/kg bw (1913 males; 1780 females). The clinical symptoms consisted of dyspnea, apathy, ataxia, abnormal positions, spastic gait, exiccosis, diarrhea, shaggy fur, saliva and a poor general state in the higher dose groups additionally diarrhea. At necropsy, findings in the animals that died comprised of redness and/or bloody ulceration of the glandular part of the stomach, redness of the mucous membrane of the stomach and gut as well as general hyperemia and bloody mucous content of the gut. No macroscopic abnormalities were observed in the surviving animals at the end of the study.

Several other acute LD50 values in rat have been determined. A study by BASF (1982) reported an oral LD50 of ca 1700 mg/kg bw in rats (males 2150; females >1210 - <1780 mg/kg bw). In these study groups consisting of 5 males and 5 females Na4EDTA was administered by gavage with 1210, 1780, 2610, 3830 mg/kg bw of a 12 - 38% solution in water. After administration a 14 day observation period followed. The clinical symptoms and pathological findings were identical to the key study performed one year later. In two prior oral LD50 determinations which were conducted the same way, oral LD 50 values of 2700 mg/kg bw (BASF 1970) and 1700 mg/kg bw (BASF, 1978) were reported.

In a study by Ciba-Geigy several groups of 5 rats per sex and dose were treated simultaneously by gavage with preparations of the test substance in 2% carboxymethyl cellulose (1470, 2150, 2400, 3170, 3590 mg/kg bw). After administration rats were observed for 7 days. The LD50 was found to be 2580 mg/kg bw (2390 - 2780 mg/kg bw). Clinical symptoms were sedation, dyspnoea, exophthalmus, curved position and ruffled fur. Additionally in the two highest dosage groups, chromodacryorrhoea and diarrhea were observed. However during necropsy nothing abnormal was observed.

An additional study by BASF was performed with a 40% Na4EDTA solution in water (Trilon B liquid). In this study groups consisting of 5 males and 5 females were administered by gavage with 3160, 4640, 6810, 10000 mg/kg bw of a 12 - 38% solution of in water. After administration a 14 day observation period followed. The oral LD50 was reported to be 3200 mg/kg bw. The clinical signs were dyspnoe, apathy, aggressiveness, prone position, ataxia, tremor, spastic gait, convulsions, exsicosis, saliva and a generally bad state.

Inhalation

In a subacute repeated dose toxicity study (BASF, 2010) 10 male Wistar rats per dose were exposed to a respirable dust aerosol

of Na2H2EDTA (CAS 139 -33 -3) 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 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

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008 

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. The oral LD50 was 1780 mg/kg bw. As a result the substance is classified for acute oral toxicity (Cat.4, H302:" Harmful if swallowed") under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation (EU) No 2016/1179.

Based on the results of a repeated dose toxicity study (5 consecutive days, see chap. 7.5.3) a classification for acute inhalation toxicity (Cat. 4; H332:"Harmful if inhaled") is done under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation (EU) No 2016/1179.

 

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