2-Butenedioic acid (2Z)-, reaction products with ammonium di-.mu.3-hydroxyhexacosa-.mu.-oxododecaoxododecatungstate(6-) (6:1), ammonium octa-.mu.-oxodi-.mu.3-oxodi-.mu.4-oxododecaoxoheptamolybdate(6-) (6:1), nickel(2+) nitrate (1:2) and nickel(2+) sulfate (1:1)

Endpoint summary

Description of key information

Oral, rat: estimated LD50: 1000 mg/kg bw (Colas, 2012)

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

Study period:

09 - 30 May 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

GROUPE INTERMINISTERIEL DES PRODUITS CHIMIQUES

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER, Le Genest St Isle, France
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: 183 - 231 g
- Fasting period before study: animals were fasted one day prior to substance administration until 4 hrs after substance administration (diet only).
- Housing: 3 animals per cage in solid-bottomed clear polycarbonate cages with stainless steel mesh lids
- Diet: M20 (pelleted), Specific Diet Services, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle:
2000 mg/kg bw: 200 mg/mL
300 mg/kg bw: 30 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw


MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 (controls), 12 (test substance group; 3 animals per step in a two-step testing procedure with 2 concentrations)

Control animals:

other: 3 animals received 10 mL/kg bw olive oil via gavage in a current control study performed from 14 Feb - 28 Feb 2012

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were observed 0.5, 1, 3 and 4 h after substance administration and daily thereafter and individual body weights were determined periodically on day 0 (prior to substance administration), 2, 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, other: macroscopical examination of oesophagus, stomach, duodenum, jejunum, ileon, caecum, colon, rectum, spleen, liver, thymus, trachea, lungs, heart, kidneys, urinary bladder, ovaries, uterus, adrenals and pancreas

Sex:

female

Dose descriptor:

LD50

Effect level:

1 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut-off according to OECD 423

Mortality:

2000 mg/kg bw: In the first step, 2/3 animals died (2 or 4 days after substance application); in the second step, 3/3 rats died (Day 2, 3 and 4)

300 mg/kg bw: No mortality was observed during the study period.

For details, please refer to Table 1 under "Any other information on results".

Clinical signs:

other: 2000 mg/kg bw: Mortalities were preceded by a decrease in muscle tone and piloerection in 2/5 and 5/5 rats, respectively. The surviving high-dose animal showed piloerection after dosing. 300 mg/kg bw: Decreased spontaneous activity and piloerection was o

Gross pathology:

2000 mg/kg bw: Non-surviving animals: Rigor mortis was noted before necropsy in 5/5 animals. Marked lysis preventing macroscopical examinations was observed in 3/5 animals. Examination of the other animals revealed thinning of the forestomach in 1/5 animals with red coloration, black coloration of the forestomach, black coloration of the corpus (1/5) and thinning of the corpus (2/5).

Surviving animals: The surviving high-dose animal revealed a thickening of the forestomach.

300 mg/kg bw: Macroscopical examinations revealed no substance-related findings.

Table 1. Mortality and clinical signs after test substance administration.

 

Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death	Mortality (%)
Females
300	3/2/6	0.5 – 3 h	---	0
2000	5/6/6	0.5 h - Day 3	Day 2, Day 3 + Day 4	83%
LD50 cut off = 1000 mg/kg bw

                                                                                           

* first number = number of dead animals                                 

 second number = number of animals with clinical signs         

 third number = number of animals used         

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: Acute Oral 4, H302
DSD: Xn, R22

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute Toxicity

Oral Route

A GLP-guideline study according to the acute toxic class method (OECD 423) was performed to determine hazardous properties of 2-Butenedioic acid (2Z)-, reaction products with ammonium di-µ3-hydroxyhexacosa-µ-oxododecaoxododecatungstate(6-) (6:1), ammonium octa-µ-oxodi-µ3-oxo-µ4-oxododecaoxoheptamolybdate(6-) (6:1), nickel(2+) nitrate (1:2) and nickel(2+) sulfate (1:1) after acute exposure (Colas, 2012). Exposure to 2000 mg/kg bw revealed mortality of 5/6 female Sprague Dawley rats which was accompanied by a decrease in body weight, muscle tone and piloerection. No mortality was observed after gavage of the next lower dose of 300 mg/kg bw. Pathological examinations of non-surviving animals showed rigor mortis prior to necropsy, marked lysis in 3/5 animals preventing macroscopical examinations and thinning of the forestomach (1/5 animals) and the corpus (2/5 animals) in the remaining animals. In contrast to the surviving high-dose animal, which revealed thickening of the forestomach, no substance – related findings were noted in low-dose animals. According to OECD 423, a LD50 cut-off value of 1000 mg/kg bw was defined in the conducted study.

Justification for selection of acute toxicity – oral endpoint
There is only one study available.

Justification for classification or non-classification

The available data on acute oral toxicity meet the classification criteria as harmful if swallowed according to Regulation (EC) 1272/2008 and Directive 67/548/EEC. No data on acute dermal and inhalation toxicity are available.