Diisopropyl adipate

Skin irritation / corrosion

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Diisopropyl adipate

EC Number:

230-072-0

EC Name:

Diisopropyl adipate

Cas Number:

6938-94-9

Molecular formula:

C12H22O4

IUPAC Name:

1,6-bis(propan-2-yl) hexanedioate

Test material form:

liquid

Details on test material:

SOURCE OF TEST MATERIAL
- Identification: Di isopropyl adipate (CAS No. - 6938-94-9)
- Source of test material: Sustainability Support Services (Europe) AB
- Appearance: Colourless Liquid
- Batch No.of test material: Lot 1/05
- Manufactured date: Not provided
- Expiration date of the lot/batch: Not provided
- Purity: 99.55%

RADIOLABELLING INFORMATION (not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored in cool place. Kept container tightly closed in a dry and well- ventilated place.
- Stability under test conditions: No data available
- Solubility and stability of the test substance in the solvent/vehicle: No data available
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data available

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No data available
- Preliminary purification step (if any): No data available
- Final dilution of a dissolved solid, stock liquid or gel: No data available
- Final preparation of a solid: No data available

FORM AS APPLIED IN THE TEST (if different from that of starting material): No data available

OTHER SPECIFICS:
- Safety precautions: Avoided contact with skin and eyes. Avoided inhalation of vapour or mist. Normal measures for preventive fire protection were taken. Aprons, caps, mask, gloves and goggles were used to ensure the health and safety of the personnel.
- Disposal: The remaining unused test item was disposed as per internal SOPs of sa-FORD and the same was documented in the raw data.

Test animals

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

Test system

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.

Results and discussion

In vivo

Results

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).

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.