Methyl ricinoleate

Endpoint summary

Description of key information

There is one key valid skin irritation study performed under OECD 404 guideline (Manciaux, 1999) and one valid eye irritation study perfomed according OECD 405 (Manciaux , 1999).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 MARCH-31 MARCH 1999

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Animals
Sex, species, strain: male New Zealand White rabbits.
Reason for this choice: species generally accepted by regulatory authorities for this type of study. Breeder: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
Number of animais: three animais were used, as recommended by the international guidelines. Identification: the animais were identified individually with a metal ear tag.
Weight: on the day of treatment, the animais had a mean body weight ± standard deviation of 2.3 ± 0.1 kg.
Acclimatization: at least 5 days before the beginning of the study.

Environmental conditions
The conditions in the animal room were set as follows:
.temperature: 18 ± 3°C
.relative humidity: 30 to 70%
. light/dark cycle: 12 h/12 h
.ventilation: approximately 12 cycles/hour of filtered, non-recycled air.
The temperature and relative humidity were under continuous control and recording. The records were checked daily and filed. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment were verified and calibrated at regular intervals.
The animais were housed individually in polystyrene cages (48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.

Food and water
During the study, the animais had free access to 112 C pelleted diet (UAR, 91360 Villemoisson­ sur-Orge, France).
Each batch of food was analysed by the supplier for composition and contaminant levels. The diet formula is presented in appendix 2.

Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analyses of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by extemal laboratories.
The results of these analyses are archived at CIT.

No contaminants were known to have been present in the diet, drinking water or bedding material at levels which may be expected to have interfered with or prejudiced the outcome of the study.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

The test substance was used undiluted.

A single dose of 0.5 ml of the test substance was placed on a dry gauze pad, which was then applied to the right flank of the animals for 4 hours.

Duration of treatment / exposure:

4 HOURS

Observation period:

The skin was examined approximately l hour, 24, 48 and 72 hours after removal of the dressing.

Number of animals:

3

Details on study design:

The test substance and the gauze pad were held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

The untreated skin served as control.
The skin was examined approximately l hour, 24, 48 and 72 hours after removal of the dressing. FollowingtheOECDandECguidelines:
Following the OECD and EC guidelines:
. when there was no evidence of dermal irritation after 72 hours, the study was ended.
. when there was persistent cutaneous irritation after 72 hours, the observation period was extended to a maximum of 14 days (until day 15) in order to determine the progress of the lesions and their reversi bili ty.
. when severe irritant effects were observed, the animais were killed on humane grounds. Any change in the animais' behaviou r was noted

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible

Table1:Individualcutaneousexaminationsandmeanvaluesofthescoresrecordedateachreading(24,48and72hours)foreachanimal

 

 

Rabbitnumber	DermalIrritation			Scores		Mean irritation	Interpretation (+)
		lhDl	24h 02	48h 03	72h 04	score(1)	(-)
716	Erythema	2	2			1.3	(-)
	Oedema	2	2	2	2	2.0	(+)

 

Other		*	*	*	*
717	Erythema	2			0	0.7	(-)
	Oedema	2	0	0	0	0.0	(-)

 

Other		*	*	*	*
718	Erythema	2	2	2	2	2.0	(+)
	Oedema	2	0	0	0	0.0	(-)

Other                    *        *        *         s

 

(l)meanofscoresondays2,3and4h       -hour

D -day

(+)-irritantaccordingtoE.E.C.criteria

(-)-non-irritantaccordingtoE.E.C.criteria

*  -None

S-Drynessoftheskin

Table1(continued)

 

 

Rabbit

Dermal

Scores

number

Irritation                                                                                                                       

 

		D5	06	D7	D8	D9
716	Erythema			0
	OedemaOther	0 •	0   s	0 •
717	Erythema
	Oedema
	Other
718	Erythema		0	0	0	0
	Oedema	0	0	0	0	0
	Other	s	s	s	s	•

 

D -day

•     -None

s-Drynessoftheskin

-Cutaneousexaminationnot performed

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Executive summary:

The studydesign was established accordingto available information on the test substance and the above OCDE 404 guideline.

 The test substance was applied for 4hours to three male New Zealand White rabbits.

 

A single dose of 0.5 ml of the undiluted testsubstance was applied to the closely-clippeds kin of one flank.

 The test substance was held in contact with the skin by means of as emi-occlusive dressing. Cutaneous reactions were observed approximately1hour,24,48 and72 hours after removal of the dressing and then daily until reversibility of cutaneous reactions.

 The mean values of the scores for erythema and oedema were calculated for each animal.

 

 Results

 

A very slight or well-defined erythema was noted in all animals from day1 up to day3 ,5 or 6 . A slight oedema was observed in all animals on day 2, it persisted up to day 4 in one of them.

Dryness of the skin was recorded in one animal on day 6and in another one between days 4 and 8.

 

Mean scores over 24, 48 and 72 hours for each animal were 1.3, 0.7 and 2.0 for erythema and

2.0 ,0.0 and 0.0 for oedema.

 

Conclusion

 

Under our experimental conditions, the test substance RICINOLEATE DEMETHYLE (batchNo.9802001) is irritant when applied topically to rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 March 1999 -4 APril 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Animais
Sex, species, strain: male New Zealand White rabbits.
Reason for this choice: species generally accepted by regulatory authorities for this type of study. Breeder: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
Number of animais: three animais were used, as recommended by the international guidelines. Identification: the animais were identified individually with a metal ear tag.
Weight: on the day of treatment, the animais had a mean body weight ± standard deviation of 2.5 ± 0.2 kg.
Acclimatization: at least 5 days before the beginning of the study.

Environmental conditions
The conditions in the animal room were set as follows:
.temperature: 18 ± 3°C
. relative humidity: 30 to 70%
. light/dark cycle: 12 h/12 h
. ventilation: approximately 12 cycles/hour of filtered, non-recycled air.
The temperature and relative humidity were under continuous control and recording. The records were checked daily and filed. ln addition to these daily checks, the housing conditions and corresponding instrumentation and equipment were verified and calibrated at regular intervals.
The animais were housed individually in polystyrene cages (48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.

Food and water
During the study, the animais had free access to 112 C pelleted diet (UAR, 91360 Villemoisson­ sur-Orge, France).
Each batch of food was analysed by the supplier for composition and contaminant levels. The diet formula is presented in appendix 2.

Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analyses of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by extemal laboratories.
The results of these analyses are archived at CIT.

No contaminants were known to have been present in the diet, drinking water or bedding material at levels which may be expected to have interfered with or prejudiced the outcome of the study.

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

A single dose of 0.1 ml of the test substance was instilled into the conjunctival sac of the left eye after gently pulling the lower lid away from the eyeball.

Observation period (in vivo):

The eyes were examined approximately 1 hour, 24, 48 and 72 hours after administration of the test substance.

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

The study design was established according to available information on the test substance and the above guidelines.

As possible irritant effects were anticipated, the test substance was administered to a single male New Zealand White rabbit, in the first instance.
Since the test substance was not irritant for this first animal, it was administered to two other male New Zealand White rabbits.

A single dose of 0.1 ml of the test undiluted substance was instilled into the left conjunctival sac. The right eye was not treated and served as control.
The eyes were not rinsed after administration of the test substance.

Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration.

The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and comeal opacity were calculated for each animal

Irritation parameter:

cornea opacity score

Basis:

other: mean 3 r

Remarks:

rabbits

Time point:

other: 24-48-72 h

Score:

0

Max. score:

0

Irritation parameter:

chemosis score

Basis:

other: mean 3 rabbits

Time point:

24/48/72 h

Score:

0 - 0

Max. score:

0

Irritation parameter:

conjunctivae score

Basis:

other: mean 3 rabbits

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

other: mean 3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

0

lndividual ocular examinations and mean values of the scoresrecorded at each reading (24,48and72hours) for each animal

 

Rabbit    Region                  Description                                  Scores                                Mean      Interpretationnumber                ofeye                    ofocular                                                                             irritation                 (+)

reactions          lh            24h          48h          72h        score(1)              (-)

Dl             02            03          04

716	Conjunctivae	Chemosis	0	0	0	0	0.0	(-)
		Redness	0	0	0	0	0.0	(-)
		Discharge	E	0	0	0	0.0
	Iris		0	0	0	0	0.0	(-)
	Comealopacity	Intensity	0	0	0	0	0.0	(-)
		Arca	0	0	0	0	0.0
	OtherFluorescein		* I	*u	* I	* I
717	Conjunctivae	Chemosis	2	0	0	0	0.0	(-)
		Redness	2	0	0	0	0.0	(-)
		Discharge	E	0	0	0	0.0
	Iris		0	0	0	0	0.0	(-)
	Comealopacity	Intensity	0	0	0	0	0.0	(-)
		Arca	0	0	0	0	0.0
	OtherFluorescein		"' I	"' u	"' I	"' I
718	Conjunctivae	Chemosis	1	0	0	0	0.0	(-)
		Redness	1	0	0	0	0.0	(-)
		Discharge	E	0	0	0	0.0
	Iris		0	0	0	0	0.0	(-)
	Comealopacity	Intensity	0	0	0	0	0.0	(-)
		Arca	0	0	0	0	0.0
	OtherFluorescein		* I	"' u	"' I	"'

 

Interpretation of results:

GHS criteria not met

Remarks:

Non irritating

Executive summary:

The study design was established according to available information on the test substance and the above guidelines.

A single dose of 0.1ml of the test undiluted substance was instilled into the left conjunctival sac.The right eye was not treated and served as control.

The eyes were not rinsed after administration of the test substance.

 

Ocular  reactions  were  observed  approximately  1hour,  24,  48  and  72 hours  after  the administration.

 

The mean values of the scores for chemosis, redness of the conjunctiva,iris lesions and comeal opacity were calculated for each animal.

 

 Results

 

Very slight reactions were observed in two animals on day 1 only; No other ocular reactions were observed during the study.

Mean scores alculated for eachanimal over 24,48 and 72hours were 0.0,0.0 and 0.0 for chemosis, 0, 0, 0, 0 and 0 for redness and 0 for iris lesions and 0 for corneal opacity.

Conclusion

Under our experimental conditions, the test substance RICINOLEATE DE METHYLE (batch No.9802001) is non-irritant when administered by ocular route to rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

In the skin irritation study (Manciaux, 1999), mean scores over 24, 48 and 72 hours for each animal were 1.3, 0. 7 and 2.0 for erythema and 2.0 ,0.0 and 0.0 for oedema. A very slight or well-defined erythema was noted in all animals from day1 up to days 3 ,5 or 6 . A slight oedema was observed in all animals on day 2, it persisted up to day 4 in one of them. Dryness of the skin was recorded in one animal on day6and in another one between days4and 8. Under our experimental conditions, the test substance RICINOLEATE DE METHYLE (batchNo.9802001) is irritant when applied topically to rabbits.

In the eye irritation study (Manciaux, 1999), very slight reactions were observed in two animals on day 1 only. No other ocular reactions were observed during the study. Mean scores calculated for each animal over 24,48 and 72hours were 0.0,0.0 and 0.0 for chemosis, 0, 0, 0, 0 and 0 for redness and 0 for iris lesions and 0 for corneal opacity.

Under our experimental conditions, the test substance RICINOLEATE DE METHYLE (batch No.9802001) is non-irritant when administered by ocular route to rabbits.

Justification for classification or non-classification

According to EU regulation (EC) No 1272/2008 (CLP) :

Skin irritation:

EU regulation (EC) No 1272/2008 (CLP), ricinoleate de methyle is classified in category 3 on skin irritation.

Justifications :mean erythema score was 1.3 (< 2).

Eye irritation:

EU regulation (EC) No 1272/2008 (CLP), ricinoletate de methyle is not classified for eye irritation.

Justifications : Cornea opacity = 0, conjunctivae score (redness) = 0, chemosis = 0 .