Diisopropyl adipate

Endpoint summary

Description of key information

Skin irritation:

The dermal irritation potential of target chemical was assessed in various in-vivo experimental studies.Based on the available studies,it can be concluded that the 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:

The ocular irritation potential of target chemical was assessed in various in-vivo experimental studies.Based on the available studies,it can be concluded that the test chemical is unable to cause eye 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''.

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report.

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

The study now reported was designed and conducted to determine the dermal Irritation/corrosion potential of test chemical in Sprague Dawley rats. This study was performed as per OECD guideline No. 402.

GLP compliance:

yes

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: In-House Bred at sa-Ford, Animal Facility
- Health Status : Healthy young adult animals were used for the study.
- Females were nulliparous and non-pregnant: yes
- Age at study initiation: No data available
- Weight at study initiation: Male: Minimum: 234 g and Maximum: 258 g Female: Minimum: 228 g and Maximum: 246 g
- Fasting period before study: No data available
- Housing: The animals were housed individually in polycarbonate cages.
- Bedding: All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) Batch No.: SPAR – 24/2013
- Room Sanitation: The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle: All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No. 400012.
- Water (e.g. ad libitum): Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period: All animals were acclimatized to the test conditions for 5 days prior to administration of the test item.
- Identification: During Acclimatization, animals were temporarily marked by permanent marker, on their tails. After acclimatization, the animals were marked by toe pad micro tattooing and cage cards. Individual cage cards were labelled with project /Study No., species, strain, sex, animal ID. and No. of animal per cage, experiment start and end date.
- Randomization: Animals were selected manually. No computer generated randomization program was used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 20.40 °C Maximum: 23.10 °C
- Humidity (%): Minimum: 38.40% Maximum: 56.00%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: From: February 10, 2014 To: March 01, 2014

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): No data available

VEHICLE
- Amount(s) applied (volume or weight with unit): No data available
- Concentration (if solution): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): No data available
- Concentration (if solution): No data available

POSITIVE CONTROL
- Amount(s) applied (volume or weight): No data available
- Concentration (if solution): No data available

Duration of treatment / exposure:

24 hours

Observation period:

14 days

Number of animals:

10 (Five per sex)

Details on study design:

TEST SITE
- Area of exposure: Dorsal area of rat skin.
- % coverage: Approx. 10% of body surface area of rat
- Type of wrap if used: Porous gauze dressing and non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test item was removed by using distilled water.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : All animals were observed once daily during days 1-14.

SCORING SYSTEM:
- Method of calculation: No data available

OTHER OBSERVATIONS
Clinical Observation
After test item administration, individual animals were frequently observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all animals were observed once a day during the 14 day observation period.

Mortality
Animals were observed twice daily for any mortality during the experimental period.

Body weight
All rats were weighed on days 0 (prior to dosing), 7 and 14.

Statistical Analysis
No statistical analysis was performed since the study was terminated with limit test.

Pathology
At the end of 14 day observation period, all the surviving rats were euthanised by overdose of CO2 and subjected to gross pathology examination, for external and internal observations.

Irritation parameter:

overall irritation score

Basis:

animal: 1 - 10

Time point:

14 d

Reversibility:

not specified

Remarks on result:

other: No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with mild erythema from day 2 to 8 and scab from day 9 to 12.

Irritant / corrosive response data:

Only female animal nos. 8 and 10 were observed with mild erythema from day 2 to 8 and scab from day 9 to 12 (refer table 2).

Other effects:

Clinical Observation
No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with erythema from day 2 to 8 and scab from day 9 to 12.

Mortality
No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period (refer table 3).

Body weight
The body weight gain was observed in male and female animals on day 7 and 14 as compared to day 0, except decline in mean body weight gain was observed in males on day 7 (refer table 1 and 4).

Pathology
The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality (refer table 5).

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Dose:2000 mg/ kg bodyweight                                                                              Density:0.9464

Animal No.	Sex	Dose (ml) Applied*	Body Weight (gram)			Body Weight Change (%)
			Day 0	Day 7	Day 14	Day 0-7	Day 0-14
1	Male	0.52	247	245	265	-0.81	7.29
2		0.49	234	215	243	-8.12	3.85
3		0.52	245	240	256	-2.04	4.49
4		0.55	258	250	261	-3.10	1.16
5		0.52	244	238	264	-2.46	8.20
6	Female	0.52	244	242	251	-0.82	2.87
7		0.52	246	246	249	0.00	1.22
8		0.49	231	235	231	1.73	0.00
9		0.48	228	229	226	0.44	-0.88
10		0.52	246	250	254	1.63	3.25

Key:* = Based on density of test item and day 0 body weight taken prior to dose application.

Table 2: Individual Animal Clinical Signs and Symptoms

 

Dose:2000 mg/kg body weight

Animal No.	Sex	Hour(s) - Day 0				Day
		1	2	3	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	Male	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
2		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
3		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
4		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
5		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
6	Female	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
7		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
8		1	1	1	1	1	65^+	65^+	65^+	65^+	65^+	65^+	65^+	146	146	146	146	1	1
9		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
10		1	1	1	1	1	65^+	65^+	65^+	65^+	65^+	65^+	65^+	146	146	146	146	1	1

Key: 1 = Normal, 65 = Erythema, 146 = Scab, + = Mild

Table 3: Individual Animal Mortality Record

 

Dose:2000 mg/kg body weight

Animal No.	Sex	Days of Observation (0 to 14)
		Morning Observations	Evening Observations
1	Male	No mortality and morbidity	No mortality and morbidity
2		No mortality and morbidity	No mortality and morbidity
3		No mortality and morbidity	No mortality and morbidity
4		No mortality and morbidity	No mortality and morbidity
5		No mortality and morbidity	No mortality and morbidity
6	Female	No mortality and morbidity	No mortality and morbidity
7		No mortality and morbidity	No mortality and morbidity
8		No mortality and morbidity	No mortality and morbidity
9		No mortality and morbidity	No mortality and morbidity
10		No mortality and morbidity	No mortality and morbidity

Table 4:Summaryof Animal Body Weight (g) and Body Weight Changes (%)

 

Dose:2000 mg/kg body weight

Sex		Body Weight (gram)			Body Weight Changes (%)
		Day 0	Day 7	Day 14	Day 0-7	Day 0-14
Male	Mean	245.60	237.60	257.80	-3.31	5.00
	SD	8.56	13.46	8.98	2.82	2.82
	n	5	5	5	5	5
Female	Mean	239.00	240.40	242.20	0.60	1.29
	SD	8.77	8.44	12.76	1.09	1.78
	n	5	5	5	5	5

Keys:SD= Standard deviation, n = Number of animals

Table 5: GrossNecropsyObservation

 

 Dose:2000 mg/kg body weight                                               Mode of Death:Terminal Sacrifice

Animal No.	Sex	Gross Observation
		External	Internal
1	Male	No abnormalities detected	No abnormalities detected
2		No abnormalities detected	No abnormalities detected
3		No abnormalities detected	No abnormalities detected
4		No abnormalities detected	No abnormalities detected
5		No abnormalities detected	No abnormalities detected
6	Female	No abnormalities detected	No abnormalities detected
7		No abnormalities detected	No abnormalities detected
8		No abnormalities detected	No abnormalities detected
9		No abnormalities detected	No abnormalities detected
10		No abnormalities detected	No abnormalities detected

Interpretation of results:

other: not irritating

Conclusions:

It was concluded that test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- Unclassified” as per CLP Classification.

Executive summary:

The study now reported was designed and conducted to determine the dermal Irritation/corrosion potential of test chemical in Sprague Dawley rats. This study was performed as per OECD guideline No. 402.

 

Five male and five female healthy young adult rats were randomly selected and used for conducting dermal irritation/ corrosion study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item was applied by single dermal application and observed for 14 days after treatment.

On test day 0,as such test item, calculated based on density (0.9464) and body weight was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area.This porous gauze dressing was covered with a non-irritating adhesive tape.The porous gauze dressing was wrapped around the abdomen and anchored with non-irritating adhesive tape.After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water.The skin reactions were assessed.

The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.

No mortality was observed in any animal till the end of the experimental period.

No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with mild erythema from day 2 to 8 and scab from day 9 to 12.

The body weight gain was observed in male and female animals on day 7 and 14 as compared to day 0, except decline in mean body weight gain was observed in males on day 7.

The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

 

Hence, it was concluded that test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- not classified” as per CLP Classification.

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 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:

TEST ANIMALS
- Source:Institute for Industrial Research & Toxicology
- Age at study initiation:10 to 12 weeks
- Weight at study initiation:2.0kg ±200g
- Housing: Rabbit was housed singly in stainless steel cages provided with stainless steel mesh bottom
- Diet (e.g. ad libitum):Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
- Water (e.g. ad libitum):Community tap water passed through ‘Aqua Guard on line water filter’ was kept in bottles, ad libitum
- Acclimation period: for 24 hours before study.

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

IN-LIFE DATES: From: To: No data available

Vehicle:

unchanged (no vehicle)

Controls:

other: The other eye, which remains untreated, served as a control.

Amount / concentration applied:

A dose of 0.1gm 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:

24 hours

Observation period (in vivo):

21 days( 1, 24, 48 and 72 hours )

Number of animals or in vitro replicates:

3 female rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): 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.
- Time after start of exposure: Afetr 24 hours. If required.

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:

72 h

Reversibility:

not specified

Remarks on result:

other: not irritating

Irritant / corrosive response data:

In the confirmatory test, the test chemical when applied to the conjunctival sac of the rabbits in the amount of 0.1 ml, did not produce any eye irritation as well as eye discharge. Furthermore, there was no other clinical sign recorded in both of the animals during the whole observation period for 21 days.

Other effects:

Irritation Scoring:
The test chemical when applied to the eye of New Zealand white rabbit at the dose level of 0.1 ml did not produce any eye irritation. Furthermore, any lesions such as pannus, staining were not recorded throughout the observation period of 72 hours.
Clinical Signs:
The test chemical applied in conjunctival sac of rabbits at the dose level of 0.1 ml 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	0×0×5=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+0+0×5=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	0×0×5=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+0+0×5=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	0×0×5=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+0+0×5=0
		B.       Chemosis	0	0	0	0	0
		C.       Discharge	0	0	0	0	0
Grand total								0.00
Mean								0.00
Eye Irritation Scoring index								0.00

Interpretation of results:

other: not irritating

Conclusions:

In the confirmatory test, the test chemical when applied to the conjunctival sac of the rabbits in the amount of 0.1 ml, did not produce any eye irritation as well as eye discharge. Furthermore, 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 was not irritating when applied in
the amount of 0.1 gm in the conjunctival sac of the rabbits under test condition.

Executive summary:

Eye irritation study was conducted as per the OECD guideline 405 to evaluate the irritation potential of the test chemical. One healthy rabbit of body weight 2.4kg 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 chemical was applied in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball at the dose rate of0.1ml.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 hoursafter 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 chemical when applied to conjunctival sac of rabbit in the amount of 0.1ml did not produce any eye irritation during the observation period. 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 of same sex and same dose level. In the confirmatory test the test compound was applied in the amount of 0.1 ml in the conjunctival sac of each 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 hoursafter 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 chemical when applied to the conjunctival sac of the rabbits in the amount of 0.1 ml, did not produce any eye irritation as well as eye discharge. Furthermore, 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 was non-irritating when applied in the amount of 0.1 gm in the conjunctival sac of the rabbits under 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:

In different studies, the test chemical has been investigated for potential for dermal irritation to a greater or lesser extent. The studies are based on in-vivo experiment conducted in human and rabbits. The predicted data using the Danish QSAR database has also been compared with the experimental data and summarized as below :

The study now reported was designed and conducted to determine the dermal Irritation/corrosion potential of test chemical in Sprague Dawley rats. This study was performed as per OECD guideline No. 402. Five male and five female healthy young adult rats were randomly selected and used for conducting dermal irritation/ corrosion study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item was applied by single dermal application and observed for 14 days after treatment.On test day 0,as such test item, calculated based on density (0.9464) and body weight was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area.This porous gauze dressing was covered with a non-irritating adhesive tape.The porous gauze dressing was wrapped around the abdomen and anchored with non-irritating adhesive tape.After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water.The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1-14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.No mortality was observed in any animal till the end of the experimental period.No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with mild erythema from day 2 to 8 and scab from day 9 to 12.The body weight gain was observed in male and female animals on day 7 and 14 as compared to day 0, except decline in mean body weight gain was observed in males on day 7.The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.Hence, it was concluded that test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- not classified” as per CLP Classification.

The above studies were supported by a dermal irritation study which 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 1.90kg 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) with the help of electric clipper one day earlier before the treatment. Animal was also observed for any dermal irritation or skin related infection at the site of application one day earlier.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. No skin reaction was observed at the site of application of test compound. 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 using two additional animals.In the confirmatory test the test compound in the amount of 0.5 ml was applied on the shaven back skin of two animals, 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 chemical in the amount of 0.5 ml did not produce any dermal irritation in terms of erythema or edema in any of the animals after the four hours application. Both the animals were also free from any clinical signs of toxicity throughout the observation period of 14 days.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".

Both the above studies were further supported by skin irritation study which was performed according to Draize method in 9 albino rabbits to assess the irritation potential of test chemical.In experiment,0.1 ml of the undiluted test chemical was applied under occlusion to the clipped back skin of nine albino rabbits. After 24 h of exposure period, the dressing was removed and the sites scored on a Primary Skin Irritation (PSI) scale of 0 (no effect) to 4 (severe erythema with or without edema).Out of 9 albino rabbits, one rabbit hardly showed perceptible erythema.The PDII score of 0.06 stipulated that the test chemical was non irritant to animal's skin.Hence,Test chemical was considered as not irritating to animal's skin after 24 hours exposure.

Furthermore, two different studies were conducted according to Patch test in 19 male and female to assess the irritation potential of test chemical.Occlusive patches were used. The undiluted test chemical was applied to the volar surface of the forearm and/or the medial aspect of arm of human volunteers at a dose 0.1 ml for 24 hours under occlusive condition.After 24 h of exposure period, the dressing was removed and the sites scored on a Primary Skin Irritation (PSI) scale of 0 (no irritation) to 4 (severe deep red erythema,vesiculation).Since the test chemical did not caused any skin lesions in both the studies, it was considered as not irritating to human skin after 24 hours exposure.

In addition with the above tests, two different studies were conducted according to Patch test in 15 male and female to assess the irritation potential of test chemical.Occlusive patches were used. The undiluted test chemical was applied to the volar surface of the forearm and/or the medial aspect of arm of human volunteers at a dose 0.1 ml for 24 hours under occlusive condition.After 24 h of exposure period, the dressing was removed and the sites scored on a Primary Skin Irritation (PSI) scale of 0 (no irritation) to 4 (severe deep red erythema,vesiculation).Since the test chemical did not caused any skin lesions in both the studies, it was considered as not irritating to human skin after 24 hours exposure. 

 

According to Danish QSAR database, Skin irritation effects were estimated by three different models i.e, Battery, SciQSAR and CASE Ultra used within Danish QSAR database for the test chemical. Based on estimation, No severe skin irritation effects were known when the test chemical was exposed to rabbit skin.

All these studies lead to a conclusion that Test chemical is indeed not irritating to skin. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical 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 experiment conducted in rabbits conducted which have been summarized as below:

Eye irritation study was conducted as per the OECD guideline 405 to evaluate the irritation potential of the test chemical. One healthy rabbit of body weight 2.4kg 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 chemical was applied in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball at the dose rate of0.1ml.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 hoursafter 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 chemical when applied to conjunctival sac of rabbit in the amount of 0.1ml did not produce any eye irritation during the observation period. 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 of same sex and same dose level. In the confirmatory test the test compound was applied in the amount of 0.1 ml in the conjunctival sac of each 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 hoursafter 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 chemical when applied to the conjunctival sac of the rabbits in the amount of 0.1 ml, did not produce any eye irritation as well as eye discharge. Furthermore, 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 was non-irritating when applied in the amount of 0.1 gm in the conjunctival sac of the rabbits under test condition.

In addition to another study, undiluted test chemical was tested for ocular irritation using six albino rabbits. One eye of each animal received 0.1 ml of the ingredient and examinations for irritation were made daily until all scores were negative or up to seven days. No irritation was caused by the test chemical. This suggests that test chemical is not irritating to the eyes of rabbits.

Both the above studies are further supported by undiluted test chemical was tested for ocular irritation using six albino rabbits. One eye of each animal received 0.1 ml of the ingredient and examinations for irritation were made daily until all scores were negative or up to seven days. On day 1 negligible irritation was caused which disappeared by day 2. This suggests that test chemical is not irritating to the eyes of rabbits.

All these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical was observed in various studies. The results obtained from these studies indicate that the test chemical is not likely 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.