Cinnamaldehyde

Endpoint summary

Description of key information

Skin irritation

The dermal irritation potential of target chemical was assessedin various in- vitro and in-vivo experimental studies which were conducted for test chemical.Based on the available key data and supporting study,it can be concluded that the testchemical is able to cause skin irritation and considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category“Category 2 (irritant)”.

 

 

Eye irritation

The ocular irritation potential of target chemical was assessedin an in- vitro and in-vivo experimental studies which were conducted for test chemical.Based on the available studies,it can be concluded thatchemical is able to cause eye irritation and considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category“Category 2 (irritant)”.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

The objective of the study was to assess the irritant and/or corrosive effects of test chemical after topical application on the intact skin in Rabbits.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:Institute for Industrial Research &Toxicology
- Age at study initiation:10 to 12 weeks
- Weight (Prior to Treatment):2.0kg±200g
- Accommodation:Animal was housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
- Diet (e.g. ad libitum):Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi
- Water (e.g. ad libitum):Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad libitum
- Acclimation period:The healthy rabbits selected for study was acclimatized to standard laboratory condition for one week in experimental room under Veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22-25°C
- Humidity (%):40-60%
- Air changes (per hr):10-15 air changes per hour
- Photoperiod (hrs dark / hrs light):12:12

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 ml

Duration of treatment / exposure:

4 hrs.

Observation period:

60 min., 24, 48 and 72 hours after application.

Number of animals:

1

Details on study design:

TEST SITE
- Area of exposure:The dorsal area of trunk
- % coverage:approximately 6 cm2.
- Type of wrap if used:impervious dressing which was secured in position with adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done):The residual test item was removed by using lukewarm water.
- Time after start of exposure:no data
SCORING SYSTEM:Draize Method

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

3.25

Reversibility:

no data

Remarks on result:

other: moderately irritant to skin of the New Zealand white rabbits

Irritant / corrosive response data:

The test compound when applied at the dose level of 0.5 ml on shaven back skin of rabbit produced focal necrotic areas, erythema and edema after four hours of patch removal. The test compound also produced deep injury to skin.

Other effects:

The test compound applied on the shaven back skin of rabbit did not produce any clinical signs of toxicity.

INDIVIDUAL ANIMAL DERMAL IRRITATION SCORES

 

Rabbit No.	Sex	INTACT SKIN
		3 Min.		4 Hours		24 Hours		48 Hours		72 Hours		14 days
		Erythema	Oedema	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema
01	F	0	0	4	2	2	1	2	1	2	0	0	0
Total		0	0	4	2	2	1	2	1	1	0	0	0
Mean		0	0	4	2	2	1	2	1	1	0	0	0
Grand Total		13.00

 

Dermal Irritation Index: 13.0/4 = 3.25

CLINICAL SIGNS

SEX	ANIMAL NO.	Time (Min.)	Time (Hours)
		3	1	4	24
FEMALE	01	N	N	N	N

N: No Clinical Signs

C: Clinical Signs Observed

Interpretation of results:

Category 2 (irritant)

Conclusions:

The dermal irritation index for test chemical was calculated as 3.25.Thus considering this irritation index score it is concluded that the test chemical is moderately irritant to skin of rabbit.

Executive summary:

The Acute Dermal Irritation study of test chemical was conducted on New Zealand white rabbit in accordance with OECD 404 to assess the irritation parameter of the test chemical.In the initial test one healthy rabbit of body weight 2.12kg selected for study after acclimatization. The test compound in the amount of 0.5 ml was applied at the different sites on the shaven back skin of animal. The hairs of back sides were removed (approximately 6 cm²) one day earlier before the treatment.The site of application was covered with impervious dressing secured in position with adhesive tape. The first patch was applied on the shaven back skin of rabbit and removed after three minutes. No serious reaction was observed at the site of application. The second patch was applied on the different shaven back side and removed after one hour. There were no signs of skin reaction observed at this site of application. Finally, a third patch was applied at a different site and was removed after four hour. Focal necrotic areas, erythema and edema were observed at the site of application at moderate level. The test was terminated after three sequential exposures due to corrosive effect of test compound.The test compound when applied at the dose level of 0.5 ml on shaven back skin of rabbit produced focal necrotic areas, erythema and edema after four hours of patch removal. The test compound also produced deep injury to skin. The test compound applied on the shaven back skin of rabbit did not produce any clinical signs of toxicity.Hence,The test compound applied on the shaven back skin of rabbit did not produce any clinical signs of toxicity.

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:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

To evaluate the acute eye irritation index of the test chemical in New Zealand White rabbit.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Age : 10 to 12 weeks
Sex : Female
Body weight range : 2.0kg ±200g
No. of animals : Three
Acclimatization : The rabbit was acclimatized to standard laboratory condition for 24 hours in experimental room before study.



ENVIRONMENTAL CONDITIONS
- Temperature (°C):22-250C
- Humidity (%):40-60%
- Air changes (per hr):10-15 air changes per hour
- Photoperiod (hrs dark / hrs light):to 12 hours artificial fluorescent light and 12 hours dark

HUSBANDRY

Accommodation:Rabbit was housed singly in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
Diet:Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
Water: Community tap water passed through ‘Aqua Guard on line water filter’ was kept in bottles, ad libitum.

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

A dose of 0.1ml of test substance was applied into the one eye of the rabbits. The other eye, which remains untreated, serves as a control.

Duration of treatment / exposure:

21 days

Observation period (in vivo):

1, 24, 48 and 72 hours

Number of animals or in vitro replicates:

Three female rabbits

Details on study design:

PREPARATION AND TREATMENT OF ANIMALS
Selection of species:
The albino rabbit is the preferable laboratory animal and only healthy young adult animals are used.
Preparation of animals:
Both eyes of each experimental animal provisionally selected and examined for any eye irritation, ocular defects, and pre-existing corneal injury within 24 hours before testing of compound.

TEST PROCEDURE:
The test compound in the amount of 0.1 ml was applied in the conjunctival sac of one eye of each animal after gentle pulling the lower eye lid away from the eye ball. The eye lids were the gently held together for about one second to prevent the loss of the material. The other eyes which remains untreated, served as control.

Irrigation:
The eyes of the test animal did not wash for at least 24 hours following instillation of the test compound. At 24 hours a washout may be used if considered appropriate.

Irritation parameter:

other: eye irritation scoring index

Basis:

animal #3

Time point:

other: 1 hr,24 hr, 498 hr72 hrs.

Score:

5

Reversibility:

not specified

Remarks on result:

other: slightly irritating

Irritant / corrosive response data:

The test compound when applied to the eye of New Zealand white rabbit at the dose level of 0.1 ml produced some blood vessels hyperemic after two hours of application of test compound upto 24 hours.

Other effects:

Clinical Signs
The test compound applied in conjunctival sac of rabbits did not show any observable clinical signs such as cage side activity and pain or stress etc. throughout the observation period of 21 days.

The eyes were examined at 1, 24, 48 and 72 hours after test substance application. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animal was observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. After recording the observations at 24 hours, the eyes were further examined with the aid of fluorescein. Individual animal weight before and during the study was also observed.

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405. The eye irritation index score is 5.0.Based on thefindings, it can be concluded that the test compound can be considered as minimally irritant when applied in the amount of 0.1ml in the conjunctival sac of the rabbits under the test condition.

Executive summary:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405.

One healthy rabbit of body weight 2.03kg was selected for study after acclimatization. Both eyes of rabbits were examined for any abnormal discharge such as eye irritation, ocular defects or pre-existing corneal injury from eye 24 hours prior to application of test compound.

The test compound was applied in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball at the dose rate of 0.1ml. The lids were then gently held for about one second in order to prevent loss of the material. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hours after the treatment. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. Further, to determine the reversibility of the effect the animal was observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weight before and during the study was observed. The test treated rabbit showed some blood vessels hyperemic after two hours of application of test compound upto 48 hours. However, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc.

 

The result obtained from the initial test was confirmed in additional two animal of same sex and same dose level (OECD-405).

 

In the confirmatory test the test compound was applied in the amount of 0.1 ml in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hours after the treatment as performed in initial test. In the confirmatory test, the treated rabbits produced some blood vessels hyperemic after two hours of application of test compound upto 48 hours. However, no other clinical signs recorded in both of the animals during the whole observation period for 21 days. The eye irritation index score is 5.0.

 

Based on above findings, it can be concluded that the test compound can be considered as minimally irritant when applied in the amount of 0.1ml in the conjunctival sac of the rabbits under the test condition.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion:

In experimental study reports, the test chemical has been investigated for potential for dermal irritation to a greater or lesser extent. The study is based on in vivo and in- vitro experiment which have been summarized as below:

The Acute Dermal Irritation study of test chemical was conducted on New Zealand white rabbit in accordance with OECD 404 to assess the irritation parameter of the test chemical.In the initial test one healthy rabbit of body weight 2.12kg selected for study after acclimatization. The test compound in the amount of 0.5 ml was applied at the different sites on the shaven back skin of animal. The hairs of back sides were removed (approximately 6 cm²) one day earlier before the treatment.The site of application was covered with impervious dressing secured in position with adhesive tape. The first patch was applied on the shaven back skin of rabbit and removed after three minutes. No serious reaction was observed at the site of application. The second patch was applied on the different shaven back side and removed after one hour. There were no signs of skin reaction observed at this site of application. Finally, a third patch was applied at a different site and was removed after four hour. Focal necrotic areas, erythema and edema were observed at the site of application at moderate level. The test was terminated after three sequential exposures due to corrosive effect of test compound.The test compound when applied at the dose level of 0.5 ml on shaven back skin of rabbit produced focal necrotic areas, erythema and edema after four hours of patch removal. The test compound also produced deep injury to skin. The test compound applied on the shaven back skin of rabbit did not produce any clinical signs of toxicity.Hence,The test compound applied on the shaven back skin of rabbit did not produce any clinical signs of toxicity.

The dermal irritation potential of test article was determined according to the OECD 439 test guideline for this study. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to the test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met and passed the acceptance of criteria.The Mean % tissue viability compared to negative control (n=3) of the test substancewas determined to be 4.1%.Hence, under the current experimental test conditions it was concluded that test substance was considered to be irritating to human skin and can thus be classified as

''Irritating to skin in Category 2” as per CLP Regulation.

Based on the available in vivo and in vitro results, the test chemical indeed have the potential to cause irritation to skin. Hence, the test chemical can be considered to be irritating to skin. Comparing the above annotations to the criteria of CLP Regulation, the test chemical can be classified under the category “category 2”.

Eye irritation:

In different studies, test chemical has been investigated for potential for dermal irritation to a greater or lesser extent. The studies are based on in- vitro and in-vivo experimental conducted in human and rabbits conducted for target chemical which have been summarized as below;

 

Acute eye irritation study was conducted to evaluate the eye irritant nature of the toxic chemical. This study was performed as per OECD guideline no. 405. Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 ml of test item was instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48, 72 hours, day 7 and 14 after test item instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of test item was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed ocular lesions, hence confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 ml of test item was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Ocular lesions were seen in animal no. 1, 2 and 3 till day 7 observations which recovered at day 14. Untreated eye of all the three rabbits was normal throughout the experimental period of 14 days. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of all 3 animals was observed at day 14. Hence under the experimental test conditions, test chemical is “An Eye Irritant (Irritating to Eyes)” to New Zealand White Female rabbit eyes. Thus, the test chemical is being classified as an eye irritant.

 

 

Another, acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405. One healthy rabbit of body weight 2.03kg was selected for study after acclimatization. Both eyes of rabbits were examined for any abnormal discharge such as eye irritation, ocular defects or pre-existing corneal injury from eye 24 hours prior to application of test compound. The test compound was applied in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball at the dose rate of 0.1ml. The lids were then gently held for about one second in order to prevent loss of the material. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hours after the treatment. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. Further, to determine the reversibility of the effect the animal was observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weight before and during the study was observed. The test treated rabbit showed some blood vessels hyperemic after two hours of application of test compound upto 48 hours. However, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc. The result obtained from the initial test was confirmed in additional two animal of same sex and same dose level (OECD-405). In the confirmatory test the test compound was applied in the amount of 0.1 ml in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hours after the treatment as performed in initial test. In the confirmatory test, the treated rabbits produced some blood vessels hyperemic after two hours of application of test compound upto 48 hours. However, no other clinical signs recorded in both animals during the whole observation period for 21 days. The eye irritation index score is 5.0. Based on above findings, it can be concluded that the test compound can be considered as minimally irritant when applied in the amount of 0.1ml in the conjunctival sac of the rabbits under the test condition.

 

 

The above in-vitro results were further supported by an-in-vitro study conducted to determine the ocular irritation potential of test article according to the OECD 492. The MatTek EpiOcular™ model was used to assess the potential ocular irritation of the test articles by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to liquid test articles and controls for ~30 minutes, followed by a ~12 minute post-soak and approximately 2 hour recovery after the post-soak. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met, passing the acceptance criteria. The mean % tissue viability of test substance was determined to be 4.1%. Hence, under the experimental test conditions it was concluded that test substance was considered to be irritating to the human eyes.

 

Based on the above summarized studies for target chemical,it can be concluded that the testchemical is able to cause eye irritation and considered as irritating.Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category“Category 2 (irritant)”.

Justification for classification or non-classification

Available data for the test chemical suggests that it is likely to cause any irritation to eyes and skin.

The test chemical can be considered to be irritating to eyes and skin and can be classified under the category “category 2 as per CLP regulation.