Aluminate(2),[[N,N'-1,2-ethanediylbis[N-(carboxymethyl)glycinato]](4-)-N,N',O,O',ON]hydroxy-,disodium

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05/02/2016 to 23/03/2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

1st addendum, 2001

Deviations:

yes

Remarks:

3 animals were between 7-8 weeks old because of limited availability of animals but this deviation did not influence the quality or integrity of the results. Prior to dosing, food was withheld for 20 to 22 hours (technical reason).

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

EPA 712-C-02-190, 12/2002

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: flakes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:040801#
- Expiration date of the lot/batch: 05.08.2016
- Purity test date:08/01/2015
- purity: 91.61%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: stable in water, instable after repeated contact to air
- Solubility and stability of the test substance in the solvent/vehicle: 100% - table in water, instable after repeated contact to air
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: no

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: dilution in aqua ad injectabilia
- Preliminary purification step (if any): a correction factor of 1.092 was applied

FORM AS APPLIED IN THE TEST (if different from that of starting material): liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River, 97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: animal n°1 and 2: 8-9 weeks
Animal n° 3-5 : 7-8 weeks
- Weight at study initiation: Animal n°1: 170 g
Animal n°2: 172 g
Animal n°3: 135 g
Animal n°4: 138 g
Animal n°5: 141 g
- Fasting period before study: between 20 and 22 hours prior testing (access to water permitted)
- Housing: The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 02102151120)
- Diet (e.g. ad libitum): Altromin 1324 maintenance diet for rats and mice (lot no. 0922) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 55 +/- 10%
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): 12 hr dark / 12 hr light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2000 mg/kg
- Amount of vehicle (if gavage): Animals n° 1/2: 2.5 ml
Animals 3/4/5: 10 ml
- Justification for choice of vehicle: chosen due to its non-toxic characteristics
- Lot/batch no. (if required): 511535 (AlleMan Pharma)

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg bw

DOSAGE PREPARATION (if unusual):
Animal 1/2: 0.546g test item into vehicle (final volume 2.5 ml)
Animal 3/4/5: 2.184g test item into vehicle (final volume 10 ml)

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: at least once during the first 30 min and with special attention during the first 4 hours post-dose. Animals were weighed on day 1 prior to the administration, day 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, other: gross pathological changes

Statistics:

Not necessary

Results and discussion

Preliminary study:

No acute oral toxicity characteristics after a single dose administration at 2000 mg/kg body weight on one animal.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

> 2 000 mg/kg bw

Based on:

test mat. (total fraction)

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No

Clinical signs:

1 animal showed spontaneous activity slightly reduced, piloerection (slight), and eyes half closed from 2 to 3h post-dosing.
1 animal showed same clinical signs from 2h to 4h post-dosing.
No specific findings in the 3 other animals.

Body weight:

Two out of 5 animals showed weight loss during the first study week but all of the animals showed weight gain during the second week.

Gross pathology:

None animals showed specific gross pathological changes during the observation period.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to the OECD 420 guideline, a single oral application of this substance to rats at a dose of 2000 mg/kg body weight was associated with minimal signs of toxicity but no mortality.
The median lethal dose of this substance after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat) above 2000 mg/kg and inferior or equal to 5000 mg/kg bw.

Executive summary:

Five female WISTAR Crl: WI(Han) rats were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The solid test item was suspended in the vehicle aqua ad injectablia (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study showing minimal signs of toxicity between 2 hours and 4 hours post-dosing.

The most relevant clinical findings were slight reduced spontaneous activity, slight piloerection and eyes half closed.

Throughout the 14-day observation period, the weight gain of the animals was within the normal range of variation for this strain except for animal no. 5 which showed a 14.9% weight loss during the first week post-dose but recovered a normal weight gain the week after.

At necropsy, no treatment-related macroscopic findings were observed in any animal of any step.

LD50:       > 2000 mg/kg ≤ 5000 mg/kg bw

Species/strain:       WISTAR Crl: WI(Han) rats

Number of animals:       5

Vehicle:       aqua ad injectabilia (sterile water)

Method:       OECD 420, Commission Regulation (EC) 440/2008