Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 August 2017 - 05 September 2017
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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
December 2001
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
December 2002
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to
Guideline:
other: Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF)
Version / remarks:
November 2000, including the most recent revisions
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Physical appearance: white powder
Storage conditions: at room temperature
Specific details on test material used for the study:
No correction factor for the purity of the test item was applied.

Test animals

Species:
rat
Strain:
other: Crl: WI(Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Animals: 9 young adult females (nulliparous and non-pregnant) of approximately 10-12 weeks old
- Weight at study initiation: 177 -219 g
- Fasting period before study: Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the test item. Water was available.
- Housing: up to 5 animals of the same sex and same dosing group were housed together in in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material equipped with water bottles. For psychological/environmental enrichment, animals were provided with paper, except when interrupted by study prcedures/activities.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
- Water: municipal tap-water was available ad libitum.
- Acclimation period: at least 5 days before the commencement of dosing.

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 18-24 (actual: 21-22)
- Humidity (%): 40-70 (actual: 49-72
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 15 August 2017 - 05 September 2017

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Remarks:
specific gravity: 1.036
Details on oral exposure:
- Justification for choice of vehicle: Trial preparations (non-GLP) were performed at the Test Facility to select the suitable vehicle and to establish a suitable formulation procedure. The vehicle is accepted by international guidelines.
- Method of application: a single dose of test item was administered to the appropriate animals by oral gavage on day 1, using a syringe with a plastic gavage cannula attached. The dosing formulations were stirred continuously during dose administration.
- Justifiction for dose level: the dose level was based on the OECD guidelines.
Doses:
2000 and 300 mg test item/kg bodyweight
Treatment was performed in a stepwise manner; a first group was treated at a dose level of 2000 mg/kg bw, based on the results two additional groups were dosed at 300 mg/kg.
No. of animals per sex per dose:
3 females per treatment group (9 animals in total)
Control animals:
no
Details on study design:
- Method: The test was performed in a stepwise treatment of 3 females, starting with a dose level of 2000 mg/kg bw. Based on the results, two additional groups were dosed at 300 mg/kg.
- Duration of observation period following administration: 14 days
- Frequency of observations:
Mortality and moribundity: twice daily
Weighing: individually on day 1 (pre-dose), day 8 and day 15.
Other examinations: at least three times on the day of dosing and once daily thereafter.
- Necropsy: yes, all moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
No statistical analysis was performed.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
500 mg/kg bw
Based on:
test mat.
Remarks on result:
other: The oral LD50 was within the range of 300-2000 mg/kg bw.
Mortality:
At 2000 mg/kg, two animals were found dead on day 1 and one animal was found dead on day 6.
At 300 mg/kg, no mortality occurred.
Clinical signs:
At 2000 mg/kg, hunched posture, flat posture, piloerection, clonic spasms, uncoordinated movements, quick breathing, shallow respiration and/or ptosis were noted between days 1 and 5 for the three animals found dead during the study.
At 300 mg/kg, hunched posture, uncoordinated movements and piloerection were noted for all animals between days 1 and 4.
Body weight:
The body weight gain shown by the majority of the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
The incidence of very slight body weight loss between days 8 and 15 for one animal treated at 300 mg/kg was considered not indicative of toxicity, based on the absence of any corroborative findings in this animal.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Remarks:
Classified as Category 4 according to Regulation (EC) No. 1272/2008
Conclusions:
The oral LD50 of HYDROPIP-NR was established in an acute oral toxicity study in rats (Acute Toxic Class Method), to be within the range of 300-2000 mg/kg bw. Based on these results, the test item is classified as Category 4 (harmful if swallowed) for acute toxicity by oral route according to the GHS and according to Regulation (EC) No. 1272/2008.
Executive summary:

An acute oral toxicity study (Acute Toxic Class method) was performed according to OECD guideline 423 and GLP principles, to determine the potential toxicity of HYDROPIP-NR. Nine female Winstar rats were dosed in a stepwise manner starting with the treatment of one group of three animals with a dose of 2000 mg/kg bw. Based on the results two additional groups of three females were treated with a dose of 300 mg/kg bw. At 2000 mg/kg, two animals were found dead on day 1 and one animal was found dead on day 6, whereas at 300 mg/kg, no mortality occurred. At 2000 mg/kg, hunched posture, flat posture, piloerection, clonic spasms, uncoordinated movements, quick breathing, shallow respiration and/or ptosis were noted between days 1 and 5 for the three animals found dead during the study. At 300 mg/kg, hunched posture, uncoordinated movements and piloerection were noted for all animals between days 1 and 4.

The mean bodyweight gain shown by the animals over the study period was considered to be normal for all treatment groups and no abnormalities were found at marcroscopic post mortem examination of the animals.

The oral LD50 value of HYDROPIP-NR in Wistar rats was established to be within the range of 300 -2000 mg/kg body weight. The LD50 cut-off value was considered to be 500 mg/kg body weight, according to OECD guideline 423.

Based on these results, HYDROPIP-NR is classified as Category 4 (harmful when swallowed) for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).