Hydrocotyle asiatica, ext.

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

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:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

2017

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Test material

Test material information

Constituent 1

Reference substance name:

Hydrocotyle asiatica, ext.

EC Number:

283-640-5

EC Name:

Hydrocotyle asiatica, ext.

Cas Number:

84696-21-9

Molecular formula:

not applicable

IUPAC Name:

Hydrocotyle asiatica, ext.

Test material form:

solid

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species/strain: WISTAR rats Crl: WI(Han)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female (non-pregnant and nulliparous)
Number of animals: 3 animals
Body weight at the
beginning of the study: 215 – 226 g
Age at the
beginning of the study: 12 - 14 weeks
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to the German Act on Animal Welfare the animals were bred for experimental purposes.
This study was performed in an AAALAC-accredited laboratory. According to German animal protection law, the study type has been reviewed and accepted by local authorities. Furthermore, the study has been subjected to Ethical Review Process and was authorised by the Bavarian animal welfare administration.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

sterile water

Duration of exposure:

24h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

1female + 2 female

Control animals:

yes

Details on study design:

Procedure Dose Range Finding Test
One animal was dosed at a dosage of 2000mg/kg body weight. As the animal survived without showing any signs of systemic toxicity, this dose was used as a starting dose for the main study.

Procedure Main Test
The dose was administered to two animals sequentially. As both animals survived at 2000 mg/kg body weight without showing any signs of systemic toxicity, no further dose was tested.

Exposure Period
The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was removed using aqua ad injectionem (sterile water).

Observation Period
All animals were observed for 14 days after dosing.

Primary Skin Irritation
Signs of erythema and oedema were assessed using the scoring system laid down in OECD Guideline 404, 24, 48 and 72 hours after patch removal

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

other: ATE

Remarks:

2000mg/kg bw

Effect level:

>= 2 000 - <= 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

0

Clinical signs:

0

Body weight:

unchanged

Gross pathology:

0

Any other information on results incl. tables

 Table: Absolute Body Weights [g] and Body Weight Change [%] - DRF and Main Study

Dose: 2000 mg/kg body weight
Animal No. / Sex	Absolute Body Weights [g]			Body Weight Change [%]
	Day 1	Day 8	Day 15	Day 1-15
1 / female	215	218	215	0
2 / female	222	220	233	5
3 / female	226	227	233	3

bw = body weight

Pathology

No specific gross pathological changes were recorded for any animal (for individual data seeTable9).

Table:  Macroscopic Findings - Individual Data – DRF and Main Study

Dose: 2000 mg/kg bw
Animal No. / Sex	Organ	Macroscopic Findings
1 / female	-	nsf
2 / female	-	nsf
3 / female	-	nsf

nsf = no specific findings

 Acute Toxicity Estimate (ATE)

Table: Acute Toxicity Estimate (ATE) according to GHS, 2017[9]

Dose (mg/kg bw)	Number of Animals Investigated (Main Study)	Number of Intercurrent Deaths	Category	ATE (mg/kg bw)
2000 mg/kg bw	3 females	0	5	2000 < ATE≤5000

bw = body weight

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present study, single dermal application of the test item Centella asiatica dry ext. to rats at a dose of 2000 mg/kg body weight was associated with no mortality and neither signs of toxicity nor signs of irritation.
According to OECD guideline 402 the test item Centella asiatica dry ext. has no obligatory labelling requirement for percutaneous toxicity and is unclassified.
The Acute Toxicity Estimate according to GHS (Globally Harmonized Classification System), 2017 was determined to be 2000 < ATE ≤ 5000 mg Centella asiatica dry ext. / kg body weight.