Sorbitan monostearate, ethoxylated

Endpoint summary

Description of key information

Skin irritation

The dermal irritation potential of test chemical was assessed in various experimental studies conducted on rabbits and human. Also the predicted data for target chemical using the Danish QSAR database has also been compared with the experimental data. Based on the available data for the key and supporting studies, it can be concluded that the test chemical is unable to cause skin irritation and thus considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

 

 Eye irritation

An ocular irritation potential of target chemical was assessed in various experimental studies conducted on rabbits. Based on the available data for the key and supporting study, it can be concluded that the test chemical is unable to cause eye damage and thus can be considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

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 report

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

To evaluate the dermal irritation potential of the test chemical in New Zealand white rabbits

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

-Source : Institute for Industrial Research & Toxicology
-Age : 10 to 12 weeks
-Body weight range : 2.0kg±200g
-Identification : By cage tag and corresponding colour body marking.
-Acclimatization : The healthy rabbits selected for study was acclimatized to standard laboratory condition for one week in experimental room under Veterinary examination.
-Randomization : After acclimatization and Veterinary examination three females were randomly selected.
-Accommodation : Animals were housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
-Diet : Pelleted feed
-Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad libitum.

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

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

0.5 gm of test compound

Duration of treatment / exposure:

4 hrs.

Observation period:

14 days

Number of animals:

Three female rabbits

Details on study design:

Details on study design
TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage:approx 6 square cm
- Type of wrap if used: impervious dressing
REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: after patch removal, the site of application was cleaned with luke warm
water
OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : 1, 24,48 and 72 hours
SCORING SYSTEM: The intact skin site of application was observed for erythema and edema and scored according to Draize method

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

8

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Skin reaction:
The test compound when applied in the amount of 0.5 gm on the shaven back skin (approximately 6 cm2) of New Zealand white rabbit did not produce any skin reaction after four hours of patch removal upto the observation period of 14 days.

Other effects:

Clinical signs:
The test compound applied on the shaven back skin of rabbit at the dose level of 0.5 gm did not produce any clinical signs of toxicity throughout the examination period of 14 days.

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	0	0	0	0	0	0	0	0	0	0
02	F	-	-	0	0	0	0	0	0	0	0	0	0
03	F	-	-	0	0	0	0	0	0	0	0	0	0
Total		0	0	0	0	0	0	0	0	0	0	0	0
Mean		0	0	0	0	0	0	0	0	0	0	0	0
Grand Total		0.00

 

Dermal Irritation Index: =0.00

TABLE – 2

CLINICAL SIGNS

 

SEX	ANIMAL NO.	Time (Min.)	Time (Hours)					Time (Day)
		3	1	4	24	48	72	14
FEMALE	01	N	N	N	N	N	N	N
	02	N	N	N	N	N	N	N
	03	N	N	N	N	N	N	N

 

N: No Clinical Signs

C: Clinical Signs Observed

 

Interpretation of results:

other: not irritating

Conclusions:

The dermal irritation index of the test chemical in New Zealand white rabbits was calculated as 0.00 and test compound can be classified under "Not Classified".

Executive summary:

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of the test chemical.

In the initial test one healthy rabbit of body weight 2.04 kg was selected for study after acclimatization. The test compound in the amount of 0.5 gm was applied uniformly 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 test substance in the amount of 0.5 gm was moistened with distilled water and then applied to a small area (approximately 6 cm2) of skin and covered with a gauze patch, which was held in place with non-irritating 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. No dermal irritation was observed at the site of application of the test compound after 4 hours of patch removal. Finally, the animal was observed for 14 days, for any irritation and corrosion.

Because no corrosive effect was observed in the initial test, a confirmatory test was done in order to confirm the irritant or negative response of the test substance by taking two additional animals. In the confirmatory test, the test compound in the amount of 0.5 gm was applied on the shaven back skin (approximately 6 cm2) of two animals (body weight of each animal was 2.13 and 2.10kg), each with one patch, for an exposure period of four hours. After four hours the patch was removed and the skin reactions were graded according to Draize’s method. The test compound applied dermally on New Zealand White rabbit in the amount of 0.5 gm did not produce any skin reaction or inflammation during the observation period. Furthermore, no clinical signs were observed during the entire observation period.

The dermal irritation index of New Zealand white rabbits is calculated as 0.00 and test compound can be classified under non- irritant category at the tested dose level of 0.5 gm.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye 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 reports

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:

-Source : Institute for Industrial Research & Toxicology
-Age : 10 to 12 weeks
-Body weight range : 2.0kg±200g
-Identification : By cage tag and corresponding colour body marking.
-Acclimatization : The healthy rabbits selected for study was acclimatized to standard laboratory condition for one week in experimental room under Veterinary examination.
-Randomization : After acclimatization and Veterinary examination three females were randomly selected.
-Accommodation : Animals were housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
-Diet : Pelleted feed
-Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad libitum.

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

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A dose of 0.1gm of test substance

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

1, 24, 48 and 72 hours after test substance application

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

Three female rabbits

Details on study design:

TEST SITE
- Area of exposure:conjunctival sac of one eye of each animal
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not washed
SCORING SYSTEM:Scale of weighted scores for grading the severity of ocular lesions developed by //Draize et al
TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein: hand-slit lamp
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.

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

110

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test compound when applied to the eye of New Zealand white rabbit at the dose level of 0.1 gm did not produce any eye irritation or any eye discharge throughout the observation period. Furthermore, any lesions such as pannus, staining were not recorded throughout the observation period of 72 hours

Other effects:

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.

TABLE- 1     GRADING OF OCULAR LESIONS

S.NO/ SEX		OBSERVATION	Score				Total	Total Score
1/F			1 hour	24hours	48 hours	72 hours
	Cornea	A.      Opacity-Degree of Density	0	0	0	0	0	0
		B.      Area of Cornea Involved	0	0	0	0	0
	Iris	A.      Values	0	0	0	0	0	0
	Conjunctivae	A.      Redness	0	0	0	0	0	0
		B.      Chemosis	0	0	0	0	0
		C.      Discharge	0	0	0	0	0
2/F	Cornea	A.      Opacity-Degree of Density	0	0	0	0	0	0
		B.      Area of Cornea Involved	0	0	0	0	0
	Iris	A.      Values	0	0	0	0	0	0
	Conjunctivae	A.      Redness	0	0	0	0	0	0
		B.      Chemosis	0	0	0	0	0
		C.      Discharge	0	0	0	0	0
3/F	Cornea	A.      Opacity-Degree of Density	0	0	0	0	0	0
		B.      Area of Cornea Involved	0	0	0	0	0
	Iris	A.      Values	0	0	0	0	0	0
	Conjunctivae	A.      Redness	0	0	0	0	0	0
		B.      Chemosis	0	0	0	0	0
		C.      Discharge	0	0	0	0	0
Grand total								0
Mean								0.00
Eye Irritation Scoring index								0.00

 

 

Interpretation of results:

other: not irritating

Conclusions:

Based on findings, it can be concluded that the test compound is practically non irritant when applied in the amount of 0.1 gm 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 1.96 kg was selected for study after acclimatization. Both eyes of rabbit 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 0.1gm. 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. 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 compound did not produce any eye irritation or any eye discharge. Furthermore, 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 animals (body weight ranges 200±gm) of same sex and same dose level .

In the confirmatory test the test compound was applied in the amount of 0.1 gm 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. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animals were 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.

In the confirmatory test, the test compound when applied to the conjunctival sac of the rabbits in the amount of 0.1 gm did not produce any eye irritation or inflammation during the entire observation period. However, there was no other clinical sign recorded in both of the animals during the whole observation period for 21 days.

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

Various studies has been investigated for the test chemical to observe the potential for dermal irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits and humans for target chemical. Also the predicted data for target chemical using the Danish QSAR database has also been compared with the experimental data that have been summarized as below;

 

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of the test chemical. In the initial test one healthy rabbit of body weight 2.04 kg was selected for study after acclimatization. The test compound in the amount of 0.5 gm was applied uniformly 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 test substance in the amount of 0.5 gm was moistened with distilled water and then applied to a small area (approximately 6 cm2) of skin and covered with a gauze patch, which was held in place with non-irritating 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. No dermal irritation was observed at the site of application of the test compound after 4 hours of patch removal. Finally, the animal was observed for 14 days, for any irritation and corrosion. Because no corrosive effect was observed in the initial test, a confirmatory test was done in order to confirm the irritant or negative response of the test substance by taking two additional animals. In the confirmatory test, the test compound in the amount of 0.5 gm was applied on the shaven back skin (approximately 6 cm2) of two animals (body weight of each animal was 2.13 and 2.10kg), each with one patch, for an exposure period of four hours. After four hours the patch was removed and the skin reactions were graded according to Draize’s method. The test compound applied dermally on New Zealand White rabbit in the amount of 0.5 gm did not produce any skin reaction or inflammation during the observation period. Furthermore, no clinical signs were observed during the entire observation period. The dermal irritation index of New Zealand white rabbits is calculated as 0.00 and test compound can be classified under non- irritant category at the tested dose level of 0.5 gm.

 

The above result was supported by the skin irritation study conducted in humans to assess the skin irritation efficacy of test chemical via Schwartz prophetic patch test.The Undiluted or 60 percent of test chemical in water were tested in 50 subject. A 72-hour occlusive patch was applied to the skin, followed after 7 days with a second 72-hour patch. No evidence of irritation was observed in any of the patients at each tested concentration. Hence the test chemical was considered to not irritating to the skin of treated subjects.

 

The potential for primary skin irritation caused by the test chemical was again evaluated using the Draize rabbit skin patch test technique.About 0.5 ml of undiluted or 15% of test chemical in water were applied and occluded for 24 hours, after which the patch sites were graded for erythema and edema on the Draize scale or another, similar scale. No evidence of skin irritation was observed at tested concentrations. Hence the test chemical was considered to not irritating to the skin of treated subjects.

 

The overall results were further supported by the predicted Danish QSAR study. According to Danish QSAR database, skin irritation effects were estimated by using four different models i.e, Battery, Leadscope, SciQSAR and CASE Ultra for test chemical. Based on estimation, no severe skin irritation effects were known when the test chemical was exposed to rabbit skin. Hence, the test chemical can be considered as not irritating to the rabbits skin.

 

Based on the available data for key and supporting studies, it can be concluded that test chemical is unable to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

 

Eye irritation:

In different studies, the test chemical has been investigated for potential for ocular irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits for target chemical that have been summarized as below;

 

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405. One healthy rabbit of body weight 1.96 kg was selected for study after acclimatization. Both eyes of rabbit 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 0.1gm. 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. 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 compound did not produce any eye irritation or any eye discharge. Furthermore, 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 animals (body weight ranges 200±gm) of same sex and same dose level. In the confirmatory test the test compound was applied in the amount of 0.1 gm 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. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animals were 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. In the confirmatory test, the test compound when applied to the conjunctival sac of the rabbits in the amount of 0.1 gm did not produce any eye irritation or inflammation during the entire observation period. However, there was no other clinical sign recorded in both of the animals during the whole observation period for 21 days. Based on above findings, it can be concluded that the test compound can be considered as practically non-irritant when applied in the amount of 0.1 gm in the conjunctival sac of the rabbits under the test condition.

 

The above result was supported by another ocular irritation study conducted for test chemical. The Draize rabbit eye irritation procedure or a modification of the test was used to evaluate the eye irritation potential of test chemical.About 0.1 ml sample was instilled into one eye of each six rabbits with no subsequent washing; some rabbits received a water wash (three rabbits received water wash). Treated eyes were examined and graded on the Draize eye irritation scale at 1, 2,3,4, and 7 days.No evidence of ocular reactions were observed at tested concentrations. Hence the test chemical was considered to not irritating to the eyes of treated subjects.

 

Thus based on the available data for the target as well as supporting study, it can be concluded that test chemical is unable to cause eye irritation and considered not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical were observed in various studies. The results obtained from these studies indicates that the chemical is unable to cause skin and eye irritation. Hence the test chemical can be classified under the category “Not Classified” for skin and eye as per CLP.