Disodium [[N,N'-ethylenediylbis[N-(carboxylatomethyl)glycinato]](4-)-N,N',O,O',ON,ON']zincate(2-)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May-June 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Disodium [[N,N'-ethylenediylbis[N-(carboxylatomethyl)glycinato]](4-)-N,N',O,O',ON,ON']zincate(2-)

EC Number:

237-865-0

EC Name:

Disodium [[N,N'-ethylenediylbis[N-(carboxylatomethyl)glycinato]](4-)-N,N',O,O',ON,ON']zincate(2-)

Cas Number:

14025-21-9

Molecular formula:

C10H12N2O8Zn.2Na

IUPAC Name:

Zinc(2+) ion disodium 2-({2-bis(carboxylatomethyl)amino]ethyl} (carboxylatomethyl) amino)acetate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Molecular formula: C10H12N2O8Zn.2Na
Molecular weight: 399.6
CAS Number: 14025-21-9
Description: White to yellowish powder
Batch: Laboratory batch
Purity: 88-95%
Test substance storage: at room temperature in the dark
Stability under storage conditions: stable
Expiry date: 15 April 2011 (allocated by NOTOX, 1 year after receipt of the test substance)
pH: 6-7 at concentration of 1%
Stability in vehicle: Water Minimum 1 week
Solubility in vehicle: Water 1000 g/L

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Source: Charles River Deutschland, Sulzfeld, Germany.
Number of animals: 6 Females (nulliparous and non-pregnant). Each dose group consisted of 3 animals.
Age and body weight: Young adult animals (approx. 8-9 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification: Earmark.

Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, temperature of 21.0 ± 3.0ºC (actual range: 19.6 - 21.4ºC), relative humidity of 40-70% (actual range: 39 - 62%) and 12 hours artificial fluorescent light and 12 hours darkness per day.

Accommodation: Group housing of 3 animals per cage in labeled Macrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom). Acclimatization period was at least 5 days before start of treatment under laboratory conditions.

Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
Water: Free access to tap water.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Method: Oral gavage, using plastic feeding tubes.
Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available.
Frequency: Single dosage, on Day 1.
Dose volume: 10 mL/kg bw
Rationale: the vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.
Preparation: the formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 females

Control animals:

no

Details on study design:

The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

Mortality/Viability: twice daily.
Body weights: days 1 (pre-administration), 8 and 15.
Clinical signs: at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
Necropsy: at the end of the 14-day observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

Not applicable (limit test)

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Hunched posture was noted for all animals on Days 1 and/or 2. Chromodacryorrhoea on Day 1 or piloerection on Day 3 were noted for individual animals.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 value of zinc-disodium EDTA in Wistar rats was established to exceed 2000 mg/kg body weight.

Executive summary:

The acute oral toxicity with zinc-disodium EDTA was examined in the rat (Acute Toxic Class Method). The study was carried out based on the guidelines described in: OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method" Commission Regulation (EC) No 440/2008, B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method" EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity" JMAFF guidelines (2000) including the most recent partial revisions. Zinc-disodium EDTA was administered by oral gavage to two groups of three female Wistar rats at 2000 mg/kg body weight (one week in between dosing of the groups). Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice 14 days after treatment. No mortality occurred. Hunched posture was noted for all animals on Days 1 and/or 2. Chromodacryorrhoea on Day 1 or piloerection on Day 3 were noted for individual animals. The body weight gain shown by the animals over the study period was considered to be normal. No abnormalities were found at macroscopic post mortem examination of the animals. The oral LD50 value of zinc-disodium EDTA in Wistar rats was established to exceed 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.