Extract of fava d'anta, obtained from the fruits of Dimorphandra mollis (Leguminosae) by solvent extraction

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25.03.2021-19.04-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Extract of fava d'anta, obtained from the fruits of Dimorphandra mollis (Leguminosae) by solvent extraction

EC Number:

953-265-8

Molecular formula:

Not applicable

IUPAC Name:

Extract of fava d'anta, obtained from the fruits of Dimorphandra mollis (Leguminosae) by solvent extraction

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: not specified
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10-11 weeks
- Weight at study initiation: 202.52 g to 224.30 g
- Housing: Maximum of three animals were housed in a standard polypropylene cage (430x285x 150 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube. For range finding study animals were housed individually during and after treatment. For main study, during treatment, the animals were housed individually; and after patch removal, animals were housed together. Clean sterilized paddy husk was provided as bedding material. Paper shredding was provided as enrichment.
- Diet: Altromin Maintenance Diet for rats and mice manufactured by Altromin Spezialfutter GmbH & Co. KG was provided ad libitum
- Water: It was provided ad libitum throughout the experimental period. Deep bore-well water passed through Reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes.The contaminant analysis test reports for the water are included
- Acclimation period: 5 (200 mg/kg body weight), 7 (1000 mg/kg body weight) and 9 days (2000 mg/kg body weight) for range finding study, and 11 days for the main study animals
- Method of randomisation in assigning animals to test and control groups: Randomization was done during acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6°C to 22.9°C
- Humidity (%): 47% to 66%
- Air changes (per hr): 12 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours fluorescent light and 12 hours dark cycle

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

Mysore Research Chemical Laboratories, Batch. No 435

Details on dermal exposure:

TEST SITE
- Area of exposure: dorso-lateral of the trunk
- % coverage: 10%
- Type of wrap if used: cotton gauze dressing and non-irritating adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with distilled water and dried with absorbent cotton
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 46.4 to 472.9 mg in Range Finding Study, 431.4 to 435.6 mg in Main Study
- Constant volume or concentration used: yes/no
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit):0.5 mL
- Lot/batch no. (if required): 435

Duration of exposure:

72 hours

Doses:

- 200 mg/kg bw (Range Finding)
- 1000 mg/kg bw (Range Finding)
- 2000 mg/kg bw (Range Finding and Main Study)

No. of animals per sex per dose:

Range Finding: 1 female per dose (3 females in total)
Main Study: 2 females per dose (2000 mg/kg bw)

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All the animals were observed for clinical signs of toxicity and mortality at 20 to 30 mins, 1 hr , 2 hrs , 4 hrs and 6 hrs (±10 mins) on treatment day 1 and thereafter once daily for clinical signs of toxicity and twice daily for mortality during the 14 days observation period. The body weights were recorded at receipt, on day 1 before test item application, on days 8 and 15.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: Observations included changes in skin, fur, eyes and mucous membranes along with changes in respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Individual animal body weights were recorded at receipt, on day 1 before test item application and on days 8 and 15 during the experimental period.
- Other examinations performed: all the animals were subjected to necropsy and a complete gross pathological examination. Histopathological examination was not carried out as there were no gross pathological findings in any of the animals.

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 treatment related mortality was observed in both range finding study and main study animals.

Clinical signs:

other: No treatment related clinical signs of toxicity were observed in both range finding study and main study animals.

Gross pathology:

No treatment related gross pathological changes were in both range finding study and main study animals.

Any other information on results incl. tables

Table 1. Clinical signs of toxicity and mortality record

Phase of the Experiment

Dose (mg/kg body weight)

Animal No.

Sex

Time of Dosing (AM)

Clinical Signs of Toxicity and Mortality on Day 1

Clinical Signs of Toxicity and Mortality on days

20-30 mins

1 hr (±10 mins)

2 hrs (±10 mins)

4 hrs (±10 mins)

6 hrs (±10 mins)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Range Finding Study

200

Rf3976

F

10:15

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

1000

Rf3977

F

10:17

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

2000

Rf3978

F

10:20

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Main Study

2000

Rf3979

F

10:32

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rf3980

F

10:34

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

 N: Normal; F: Female; mins: minutes; hr/hrs: hour/hours

 

Table 2. Individual animal local skin reactions

Phase of the Experiment	Animal No	Sex	Dose (mg/kg body weight)	Days
				3 (24 hours)		4 (48 hours)		5 (72 hours)
				ER	ED	ER	ED	ER	ED
Range finding Study	Rf3976	Female	200	0	0	0	0	0	0
	Rf3977	Female	1000	0	0	0	0	0	0
	Rf3978	Female	2000	0	0	0	0	0	0
Main Study	Rf3979	Female	2000	0	0	0	0	0	0
	Rf3980	Female	2000	0	0	0	0	0	0

ER: Erythema; ED: Edema; 0: No erythema/edema

 

Table 3. Body weight (g) and percent change in body weight with respect to day 1

Phase of the Experiment	Dose (mg/kg body weight)	Animal No.	Sex	Quantity Administered (mg)	Body Weight (g) on Days				Percent Change in Body Weight with Respect to Day
					1	8	15		1 to 8	1 to 15
Range Finding Study	200	Rf3976	F	46.4	231.93	246.12	261.28		6.12	12.65
	1000	Rf3977	F	232.7	232.68	246.21	260.02		5.81	11.75
	2000	Rf3978	F	472.9	236.47	249.83	264.21		5.65	11.73
Main Study	2000	Rf3979	F	431.4	215.72	230.31	245.20		6.76	13.67
		Rf3980	F	435.6	217.78	233.14	248.12		7.05	13.93
		Mean			216.75	231.73	246.66		6.91	13.80
		±SD			1.46	2.00	2.06		0.20	0.19
		n			2	2	2		2	2

F: Female; SD: Standard Deviation; n: number of animals.

 

Table 4. Gross pathology findings

Phase of the Experiment	Dose (mg/kg body weight)	Animal No.	Sex	Fate	Gross Pathology Findings
					External	Internal
Range finding Study	200	Rf3976	F	TS	NAD	NAD
	1000	Rf3977	F	TS	NAD	NAD
	2000	Rf3978	F	TS	NAD	NAD
Main Study	2000	Rf3979	F	TS	NAD	NAD
		Rf3980	F	TS	NAD	NAD

NAD: No Abnormality Detected; F: Female; TS: Terminal Sacrifice

Applicant's summary and conclusion

Interpretation of results:

other: Not classified (CLP Regulations EC no. 1272/2008)

Conclusions:

The LD50 value of the test item is >2000 mg/ kg body weight by dermal route in rat.

Executive summary:

The acute dermal toxicity of the test item has been performed in accordance with OECD Test Guideline 402, following GLP. A thin paste of the test item and water was applied to a total of 5 female Sprague-Dawley rats in two differents studies. A range finding study using three female rats was performed using a concentration of 200, 1000 and 2000 mg/kg b.w., respectively. As no clinical signs of toxicity nor mortality were observed at any of these concentrations, the main test was carried out using 2 more female rats at a concentration of 2000 mg/kg b.w. Again, no clinical signs of toxicity nor mortality were observed. No adverse changes were observed in body weight and percent change in body weight with respect to day 1 and no gross pathological changes were observed in any of the studies performed. Therefore, the test item has a LD50 value of >2000 mg/kg b.w. As such, the test item is not classified acccording to CLP Regulations EC no. 1272/2008.