Pregnenolone

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 July 2017 - 11 August 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

May 2008

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

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:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Pregnenolone

EC Number:

205-647-4

EC Name:

Pregnenolone

Cas Number:

145-13-1

Molecular formula:

C21H32O2

IUPAC Name:

3-hydroxypregn-5-en-20-one

Test material form:

solid: particulate/powder

Details on test material:

- Physical appearance: white crystalline powder
- Storage conditions: at room temperature

Specific details on test material used for the study:

- No correction factor for purity required.
- pH: 6.25 - 5.93

Test animals

Species:

rat

Strain:

other: Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Animals: 6 young adult females (nulliparous and non-pregnant) of approximately 10 weeks old
- Weight at study initiation: 176 -197 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
- Temperature (°C): 21 to 22°C
- Humidity (%): 46-69
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 25 July 2017 - 11 August 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.
- 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 mg test item (10 mL) /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 a second group was dosed at 2000 mg/kg.

No. of animals per sex per dose:

3 females per treatment group (6 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, an additional group was dosed at 2000 mg/kg bw.
- 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 (the method used is not intended to allow the calculation of a precise LD50 value).

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 occurred.

Clinical signs:

other: Hunched posture, piloerection and/or uncoordinated movements were noted for the animals on Day 1. Additionally, lethargy was noted in a single animal on Day 1.

Gross pathology:

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

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Not classified according to Regulation (EC) No. 1272/2008

Conclusions:

The oral LD50 of PRECH-PURCH was established in an acute oral toxicity study in rats (Acute Toxic Class Method), to exceed 2000 mg/kg bw. Based on these results, the test item salt does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the GHS.

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 PRECH-PURCH. Six female Winstar rats were dosed with a single limit dose of 2000 mg/kg bodyweight. The study was performed in a stepwise manner: a first group of 3 animals was treated at a dose level of 2000 mg/kg bw, based on the results a second group of 3 animals were treated. No mortality occurred. Hunched posture, piloerection and/or uncoordinated movements were noted for the animals on day 1. Additionally, lethargy was noted in a single animal on day 1. The mean bodyweight gain shown by the animals over the study period was considered to be normal and no abnormalities were found at marcroscopic post mortem examination of the animals.

The oral LD50 value of PRECH-PURCH in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, PRECH-PURCH does not have to be classified and has no obligatory labelling requirement 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).