Willow, Salix alba, ext.

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05.02.2019 - 09.04.2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

α-hydroxy-o-tolyl β-D-glucopyranoside

EC Number:

205-331-6

EC Name:

α-hydroxy-o-tolyl β-D-glucopyranoside

Cas Number:

138-52-3

Molecular formula:

C13H18O7

IUPAC Name:

(2R,3S,4S,5R,6S)-2-(hydroxymethyl)-6-[2-(hydroxymethyl)phenoxy]oxane-3,4,5-triol

Test material form:

solid: particulate/powder

Details on test material:

Name: White willow bark extract
CAS number: 84082-82-6

Specification:
Purity: 100% (UVCB substance)
Appearance: Brown powder
Odour: Characteristic
Sieve analysis: 100% pass 80 mesh

Loss on drying: 5.0 % max
Residue on ignition: 5.0 % max
Bulk density: 40 – 55 g/100ml
Extract solvent: Alcohol & water
Heavy metal: 10 ppm max
Lead (Pb): 2 ppm max
Arsenic (As): 2 ppm max
Cadmium (Cd): 2 ppm max
Mercury (Hg): 2 ppm max
Residual solvents: Complies with Eur.Pharm.


Microbiological:
===============
Total Plate Count: 1000 cfu/g max
Yeast & Mould: 100 cfu/g max
E. coli: Negative
Salmonella: Negative

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: ~10 weeks
- Weight at study initiation: 210 - 230 g
- Fasting period before study: fasting since night before treatment
- Housing: Group caging (3 animals/cage)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 19 or 20 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.4 – 22.7 °C
- Humidity (%): 25 – 74 %
- Air changes (per hr): 12-20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: default vehicle with appropriate solubility
- Purity: not applicable

MAXIMUM DOSE VOLUME APPLIED:


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: A limit test with 2000 mg/kg bw was performed as no toxicity was expected based on knowledge of consituents

Doses:

2000 mg/kg bw (limit test)

No. of animals per sex per dose:

6. 3 in inital test, 3 in confirmatory test.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical observations 0.5, 1, 2, 3, 4, 6 h after treatment and daily thereafter. Weighing weekly and at necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: macroscopic observation of organs and tissues

Statistics:

Not applicable (limit test, no mortality)

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The test item did not cause mortality at a dose level of 2000 mg/kg bw in any animal (2 groups of 3 females)

Clinical signs:

other: There were no systemic clinical signs noted in any animal throughout the study

Gross pathology:

There was no evidence of the macroscopic changes in any animal.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item, White willow bark extract was found to be above 2000 mg/kg bw in female Crl:WI Wistar rats.

According the GHS criteria, classification of the test item can be ranked as "Category 5 or Unclassified" for acute oral exposure.

Executive summary:

The single-dose oral toxicity study with the test item was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris) in Crl:WI Wistar female rats in compliance with GLP.

Two groups of three female Crl:WI rats were treated with the test item at a dose level of 2000 mg/kg body weight (bw) (Group 1 and Group 2).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was administered at the dose level of 2000 mg/kg bw.

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory 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.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 7 and before necropsy (Day 14). All animals were subjected to a necropsy and a macroscopic examination.

The test item did not cause mortality at a dose level of 2000 mg/kg bw.

There were no systemic clinical signs noted in any animal throughout the study. There were no treatment related body weight changes. The body weights of the treated animals were within the range commonly recorded for this strain and age.

There was no evidence of the macroscopic changes at a dose level of 2000 mg/kg bw.