Cyperus scariosus, ext.

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

6th February 2018-22nd March 2018

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

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Cyperus scariosus, ext.

EC Number:

294-955-2

EC Name:

Cyperus scariosus, ext.

Cas Number:

91771-62-9

IUPAC Name:

Cyperus scariosus, ext.

Test material form:

liquid

Specific details on test material used for the study:

Batch Manufactured and supplied by
Jagat Aroma Oils Distillery,
Kannauj-209725,
Uttar Pradesh, India

Test Item : CYPRIOL (Cyperus Scariosus ext. Oil)

CAS No. : 91771-62-9 / 68916-60-9

Chemical Name (IUPAC)
Not applicable

Physical Appearance : Amber colored or Light Brown Liquid

Purity as per Certificate of Analysis: 100% Natural

Batch No. : Lot # 11/CYP/2016

Manufactured Date : November 2016

Retest Date : October 2018

Recommended Storage Condition: Ambient (+15 to +25°C)

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: In-Vivo Biosciences, Kodigehalli Village, Magadi Road, Bangalore, Code-29, Karnataka State

- Females (if applicable) nulliparous and non-pregnant: yes

- Age at study initiation: 9 to 11 weeks

- Weight at study initiation: 189.5-197.8g

- Fasting period before study: 16-18 hours

- Housing: Rats were housed individually in standard polysulfone cages
(Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottle. Additionally, polycarbonate rat huts were placed inside the cage as an enrichment object and were changed along with the cage once a week.

Bedding: steam sterilized corn cob was used and changed once a week along with the cage.

- Diet (e.g. ad libitum): yes. Hypro Rat & Mice pellet feed, manufactured by Krishna Valley Agro Tech LLP, MIDC Kupwad block, Sangli, Maharashtra, was provided to animals.

- Water (e.g. ad libitum): yes. Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd, Mumbai 400 001, India, was provided to animals in polycarbonate bottles with stainless steel sipper tubes.

- Acclimation period: After physical examination for good health and suitability for experiment, the animals were acclimatized for six, eight, ten and twelve days before treatment G1-FTS, G1-STS, G2-FTS and G2-STS respectively. Animals were observed once daily during acclimatization period. Females were nulliparous and non-pregnant.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24 °C
- Humidity (%): 64 to 67 %
- Air changes (per hr): 13.2 to 14.0 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The undiluted test item as supplied by the sponsor was administered as a single oral gavage to the fasted (16 to 18 hours) access to water was not interrupted. Each animal was administered the test item orally by gavage using disposable plastic syringe attached with metal feeding cannula. Food was offered about 3 to 4 hours after dosing. Water was not withheld

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: There was no acute oral toxicological information is available for the test item Cyperus Scariosus. Hence, the study was initiated with the starting dose of 300 mg/kg body weight. Hence the test was started as per Annex 2c of the OECD 423 test guideline. The starting dose was 300 mg/kg body weight (G1 FTS).

Doses:

300 & 2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

Group Dose
(mg/kg) No. of
Rats Sex Rat numbers
From To
G1 (FTS) 300 3 Female Rm8031 Rm8033
G1 (STS) 300 3 Female Rm8034 Rm8036
G2 (FTS) 2000 3 Female Rm8037 Rm8039
G2 (STS) 2000 3 Female Rm8040 Rm8042

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

G1 - [300 mg/kg body weight – (FTS and STS)]: There were no pre-terminal deaths.
G2 - [2000 mg/kg body weight - (FTS and STS)]: There were no pre-terminal deaths.

Clinical signs:

other: G1 - [300 mg/kg body weight – (FTS and STS)]: There were no clinical signs. G2 - [2000 mg/kg body weight - (FTS and STS)]: There were no clinical signs.

Gross pathology:

No abnormalities were detected at necropsy.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the present study, the test item, CYPRIOL (Cyperus Scariosus ext. Oil) has oral LD50 in the range of 2000 to 5000 mg/kg. The test item does not meet the criteria for classification per Regulation (EC) No 1272/2008.

Executive summary:

The acute oral toxicity study with CYPRIOL (Cyperus Scariosus ext. Oil) in Wistar rats was conducted to assess the toxicological profile of the test item.  The undiluted test item was administered as a single oral gavage to overnight fasted (16 to 17 hours) three female rats (G1-FTS) at a dose volume of 0.31 mL/kg body weight to attain the dose of 300 mg/kg body weight (G1-FTS). There were no clinical signs and pre-terminal deaths.

Based on the scheme - Annex 2c of the guideline OECD 423, three additional female rats were tested at the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs and pre-terminal deaths. Based on the scheme - Annex 2c of the guideline OECD 423, three female rats were tested at the next higher dose of 2000 mg/kg body weight and at the dose volume of 2.06 mL/kg body weight (G2-FTS). There were no clinical signs and pre-terminal deaths. Based on the scheme - Annex 2c of the guideline OECD 423, additional three female rats were tested at the same dose of 2000 mg/kg body weight (G2-STS). There were no clinical signs and pre-terminal deaths. Based on the scheme - Annex 2c of the guideline OECD 423, the dose was stopped.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Necropsy was performed for all the survived rats at terminal sacrifice. There were no clinical signs and mortality during the experiment period. All rats gained weight during experimental period. The rats were subjected to necropsy at termination and there were no abnormalities detected at necropsy.