Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-01-09 to 2018-03-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
adopted 17 December 2001
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
- CAS number: 39464-70-5
- Appearance: Clear, yellow-amber liquid
- Purity: 100%
- Storage conditions: Controlled room temperature (15-25 °C, ≤ 70% relative humidity (RH))

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle, in the Pharmacy of Citoxlab Hungary Ltd. on the day of administration. The formulation container was stirred continuously up to finishing the treatment

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Young healthy adult rats, 12 weeks old
- Weight at study initiation: 215 – 244 g
- Fasting period before study: Night before the treatment
- Housing: 3 animals / cage
- Diet (e.g. ad libitum): Yes, ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany (Batch no.: 382 24962, expiry date: 30 April 2018)
- Water (e.g. ad libitum): Yes , tap water
- Acclimation period: at least 32 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 -22.5
- Humidity (%): 36-83
- Air changes (per hr): 15 – 20
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Justification for choice of vehicle: On the basis of the trial formulations with the test item, the vehicle used was distilled water.
- Lot/batch no. (if required): 8130917

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: A limit dose of 2000 mg/kg bw was selected by the Sponsor. Initially, three females (assigned to Group 1) were treated at a dose level of 2000 mg/kg bw. The test item did not cause mortality in this group and a second group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required according to OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Frequency of weighing: Days -1, 0 and 7 and Day 14
- Necropsy of survivors performed: All animals were subjected to a necropsy and a macroscopic examination. The animals were exsanguinated after verification of narcosis following an injection of pentobarbital sodium (Euthanimal 40%; Lot No.: 1609291-03, Expiry Date: October 2019, Produced by: Alfasan International B.V.). After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross macroscopic changes were recorded for each animal.
Statistics:
Mean and standard deviation

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed
Clinical signs:
Slight clinical signs were noted in animals treated at the dose level of 2000 mg/kg bw with Ethoxylated phenol phosphate. These included decreased activity (2 / 6), hunched back (6 / 6), incoordination (6 / 6), and piloerection (6 / 6). The clinical signs reversed by day 2.
Body weight:
No treatment related effects were observed
Gross pathology:
No treatment related effects were observed

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In an acute oral toxicity study in rats conducted according to OECD 423 no mortality occurred at the limit dose of 2000 mg/kg bw. Hence, the LD50 value was determined to be greater than 2000 mg/kg bw.
Executive summary:

In an acute oral toxicity study (acute toxic class method, OECD 423), two groups of fasted, 12 weeks old, female Wistar rats (3 rats/group) were given a single oral dose of the test item (100% purity) in water at the limit dose of 2000 mg/kg bw and were observed for 14 days. All animals survived until the end of the study showing only mild signs of toxicity. The clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, hunched back, incoordination and piloerection. All symptoms were no longer detectable by day 2 after dosing.

Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain. At necropsy, no macroscopic findings were observed in any animal of any step. Based on the results from this study, the oral LD50 in rats is considered to exceed 2000 mg/kg bw.