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Administrative data

Description of key information

Acute oral toxicity - Weight of evidence suggests the median lethal dose to be close to 2000 mg/kg. In the absence of reliable data on the substance itself a key, well documented study, on the analogue tetrasodium EDTA describes the LD50 of that substance as being 1780 mg/kg. This is used to characterise the acute toxicity of this substance and the LD50, re-based according to the molecular weight of the two substances, is therefore taken as being 1303 mg/kg.
Acute inhalation toxicity - The LC50(4h) for the structurally related substance Na3-HEDTA is in excess of 3.95 mg/L, the highest exposure concentration achievable.
Acute dermal toxicity - No reliable data available.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
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:
other: Study meets generally accepted scientific principles (non-GLP) For read-across justification refer to section 13.
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
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals 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
Sex:
male
Dose descriptor:
LD50
Effect level:
1 913 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
1 780 mg/kg bw
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 780 - < 2 000 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
Interpretation of results:
harmful
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The observed acute oral toxicity indicates that the substance should be classified with R22 according to EU criteria and acute toxicity category 4 according to GHS criteria.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
1 303 mg/kg bw
Quality of whole database:
A number of studies are available regarding acute oral toxicity providing a weight of evidence that suggests that the LD50 is close to 2000 mg/kg. In the absence of reliable data on the substance itself a key, well documented study, on the analogue tetrasodium EDTA describes the LD50 of that substance as being 1780 mg/kg. This is used to characterise the acute toxicity of this substance and the LD50, re-based according to the molecular weight of the two substances, is therefore taken as being 1303 mg/kg.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
05 November 2012 - 04 December 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline Study (according to OECD 403)
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories B.V., Kreuzelweg 53, 5961 NM Horst, Niederlande
- Age at study initiation: males approximately 8-9 weeks, females approximately 10-11 weeks
- Weight at study initiation (g): Group 1 males = 229.4 +/- 5.7, Control males = 233.4 +/- 5.1, Group 1 males = 231.5 +/- 8.6, Group 1 females = 199.7 +/- 5.5, Group 2 males = 260.1 +/- 3.0, Group 2 females = 215.0 +/- 4.9
- Fasting period before study: not reported
- Housing: single housing or up to 5 animals in Makrolon cages, type M III (floor area about 800 cm^2) for single housing, or Polysulfon cages (H-Temp [PSU]), floor area about 2065 cm^2 (610 x 435 x 215 mm); supplied by TECNIPLAST, HohenpeiBenberg, Germany when caged in groups, if the animals were free from clinical signs and finding
- Enrichment: wooden gnawing blocks (Typ NGM E-022): Abedd Lab and Vet Service GMBH Vienna, Austria
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days before exposure
- Identification: individual identification by cage cards and tail markings

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 degrees C
- Humidity (%): 30-70 % relative humidity
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 03 November 2012 To: 04 December 2012
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose/head only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: head-nose inhalation system INA 20 (glass-steel construction, BASF SE, volume V ~ 55 L)
- Exposure chamber volume: 55 L
- Method of holding animals in test chamber: restrained in glass tubes with their snouts projected into the inhalation system
- Source and rate of air: compressed conditioned air at 1.5 m^3/h
- Method of conditioning air: central air conditioning system provides cold air of about 15 degrees C. This cold air will pass through an activated charcoal filter, be adjusted to room temperature of 20 to 24 degrees C and pass through a second particle filter (H13 (HEPA) Camfil Farr, Germany). The so generated conditioned air is used to generate inhalation atmospheres.
- System of generating particulates/aerosols: aerosols were produced by continuously pumping amounts of the test substance to a two-component atomizer. Using compressed air, the aerosol was produced with the atomizer inside the exposure system.
- Method of particle size determination: vacuum sample from the breathing zone 30 minutes after the beginning of the exposure, the amounts of material adsorbed to the walls of the impactor and in the sampling probe were also determined quantitatively. The particle size distribution was calculated based on the non-volatile fraction of test substance.
- Treatment of exhaust air: filtered and conducted into the exhaust air of the building
- Temperature, humidity, pressure in air chamber: about 21 degrees C, 40-70% humidity, 1.5 bar pressure

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric measurement of the inhalation atmosphere concentration was used
- Samples taken from breathing zone: yes

VEHICLE - Not used

TEST ATMOSPHERE: See tables below
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
0, 1.963 mg/L and 10.054 mg/L (maximum achievable concentration) - based on substance as supplied.
0, 1.003 mg/L and 5.138 mg/L (maximum achievable concentration) - based on non-volatile fraction (51.1%)
0, 0.77 mg/L and 3.95 mg/L (maximum achievable concentration) - based on substance itself (39.3% concentration in formulation)
No. of animals per sex per dose:
5
Control animals:
yes
Details on study design:
- Duration of observation period following administration: at least 14 days
- Frequency of observations and weighing: detailed clinical observations were recorded for each animal separately several times during exposure and at least once daily on the pre-exposure day and during the observation period. Individual body weights were once during the acclimatization period, shortly before exposure (day 0), and at least on days 1, 3, and 7, and before the sacrifice of the animals at the end of the observation period.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, histopathology
- To evaluate morphological changes of respiratory tract, a satellite group (5 male rats) was exposed to the test substance simultaneously with the main group animals.
Statistics:
For weight parameters, non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each test group with the control group was performed using the WILCOXON test for the hypothesis of equal medians. Binomial test was used for statistical evaluation for mortality.
Preliminary study:
Not applicable
Sex:
male/female
Dose descriptor:
LC0
Effect level:
> 10.054 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC0
Effect level:
5.138 mg/m³ air (analytical)
Based on:
other: Non-volatile fraction
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC0
Effect level:
3.95 mg/L air (analytical)
Based on:
act. ingr.
Remarks:
HEDTA at 39.3% concentration
Exp. duration:
4 h
Mortality:
Test group 1 (1.963 mg/L): No lethality was observed in male and female animals during the study period of 14 days.
Test group 2 (10.054 mg/L): No lethality occurred in male and female animals during the study period of 14 days.
Controls: No lethality was reported in the male or female group.
Clinical signs:
No abnormalities were detected in the control animal of the satellite group during the study period.
For the exposed animals, Tables 2-4 (below) show the duration of signs. No abnormalities were detected in the test group 1 animals during the post exposure observation period from study day 3 onwards. No abnormalities were detected in the test group 2 animals during the post exposure observation period from study day 7 onwards.
Body weight:
Control animals: The minimally body weight decreased from exposure day to study day 1 was most likely to be a result of food and water withdrawal during exposure.
Test group 1 (1.963 mg/L): Satellite animals: The mean body weights of the animals decreased during the first post exposure observation day.
Main group animals: The mean body weights of the animals decreased during the first post exposure observation day and increased from study day 3 onward.
Test group 2 (10.054 mg/L): The mean body weights of the animals decreased during the first post exposure observation day and increased from study day 3 onward.
Gross pathology:
No relative organ weight changes were recorded for any group.
No macroscopic lesions were recorded for any group.
In the Test group 1 satellite group, the larynx findings are summarized in Table 6 below. Overall, The necrosis of larynx was characterized by a multifocal to coalescing loss of all epithelial layers with hemorrhage and fibrin exudation as well as infiltration of inflammatory cells.
Other findings:
Not applicable

Table 2. Duration of signs, test group 1, main group (1.963 mg/L).

 

Clinical Sign

Male Animals

Female Animals

Fur, substance contaminated

d0

d0

Piloerection

d0

d0 – d1

Respiration, labored

h3 – h4

h3 – h4

Respiration, sounds

d0 – d2

d0 – d2

Table 3.  Duration of signs, test group 2 (10.054 mg/L).

 

Clinical Sign

Male Animals

Female Animals

Fur, substance contaminated

d0 – d4

d0 – d4

Nose, red encrusted

d0 – d1

--

Piloerection

d0 – d4

d0 – d4

Respiration, labored

h1 – h4

h1 – h4

Respiration, sounds

d0 – d6

d0 – d6

Table 4. Duration of signs, test group 1, satellite group (exposed for 4 hours at 1.963 mg/L and killed after 24 hours)

Clinical Sign

Male Animals

Fur, substance contaminated

d0

Piloerection

d0 – d1

Respiration, labored

h3 – h4

Respiration, sounds

d0 – d1

Table 5. Particle size analysis

Test Group

Sample

MMAD (um)

Geometrical Standard Deviation

1

1

0.6

6.8

1

2

1.6

4.9

2

1

2.0

3.1

2

2

1.5

3.5

MMAD - mass median aerodynamic diameter) is the calculated aerodynamic diameter which divides the size distribution in half when measured by mass

Table 6. Group 1 Satellite (5) Males, summary of larynx histopathology.

Larynx

Test Group

 

 

0 (Control)

1 (1.963 mg/L)

Organs Examined

5

5

Level II

 

 

 Necrosis, (multi)focal

0

5

·  Grade 1

 

1

·  Grade 3 

 

2

·  Grade 4

 

2

 Infiltration, inflammatory cells

0

5

·   Grade 1

 

1

·   Grade 2

 

1

·   Grade 3

 

3

Level III

 

 

 Necrosis, (multi)focal

0

5

·   Grade 2

 

2

·   Grade 3

 

2

·   Grade 4

 

1

 Infiltration, inflammatory cells

 

5

·   Grade 2

 

4

·   Grade 3

 

1

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the current study conditions the LC50 for male and female rats after liquid aerosol inhalation exposure of Masquol EDTA OH for 4 hours was estimated to be > 10.054 mg/L (> 3.95 mg/L HEDTA), this being the maximum technically achievable concentration.
Executive summary:

The acute inhalation toxicity (single 4 -hour exposure) of Masquol EDTA OH has been investigated in the rat according to OECD, EU and US EPA test guidelines and using an aerosol generated from the liquid formulation.

The following measured concentrations were tested:

0 (control), 1.963 mg/L and 10.054 mg/L expressed as the supplied formulation

0 (control), 1.003 mg/L and 5.138 mg/L expressed as the non-volatile fraction of the supplied formulation (51.1%)

0 (control), 0.77 mg/L and 3.95 mg/L expressed as the active substance content of the supplied formulation (39.3% HEDTA)

The higest concentration investigated was the maximum technically attainable with the test system and particle size distributions (MMAD) was between 0.6 and 2.0 μm, within the respirable range.

No lethality was observed in male and female animals in both concentrations during the study period of 14 days and the LC50 is therefore in excess of the maximum concentration achievable, 10.054 mg/L supplied formulation / 3.95 mg/L HEDTA.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
3 950 mg/m³
Quality of whole database:
Single study available on the the structurally related substance Na3-HEDTA.

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
No reliable studies available

Additional information

A number of studies is available regarding acute oral toxicity providing a weight of evidence that suggests that the LD50 is close to 2000 mg/kg. In the absence of reliable data on the substance itself a key, well documented study, on the analogue tetrasodium EDTA describes the LD50 of that substance as being 1780 mg/kg. This is used to characterise the acute toxicity of HEDTA-H3 and the LD50, re-based according to the molecular weight of the two substances, is therefore taken as being 1303 mg/kg.

 

An inhalation study on the structurally related substance HEDTA-Na3 indicates the LC50 to be in excess of 3.95 mg/L following 4 hours exposure, this being the highest exposure concentration achievable. An acute inhalation toxicity study with EDTA-Na2H2 showed a 6 -h LC30 value of 1000 mg/m3 and was thus more toxic than HEDTA-Na3, probably due to the higher binding capacity of EDTA compared to HEDTA.

 

No reliable data are available on dermal toxicity but the expected low dermal absorption of the substance does not indicate a concern for acute effects.


Justification for selection of acute toxicity – oral endpoint
Well documented study on the analogue tetrasodium EDTA.

Justification for classification or non-classification

Taking into account the provisions laid down in CLP (1272/2008/EC), classification as acute toxicity category 4 (H302) according to GHS criteria is required.