1,2-Cyclohexanedicarboxylic Acid, 1-(phenylmethyl) ester, ester with 2,2,4-trimethyl, 1,3-petanediol mono(2-methyl propanoate)

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 June 2015 to 16 September 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

The pretest body weight of Animal 5 exceeded the +/- 20% of the mean body weight by 2 grams. No impact is expected since the degree of weight deviation is negligble. Each animal received a dose based on their individual body weight.

Principles of method if other than guideline:

Deviation to the Protocol
The pretest body weight of Animal 5 exceeded the +/- 20% of the mean body weight by 2 grams. No impact is expected since the degree of weight deviation is negligible. Each animal received a dose based on their individual body weight.

Amendment to the Protocol
At the request of the Sponsor, additional analysis was conducted according to the current Globally Harmonized System of Classification and Labeling of Chemicals.
The animal supplier, Charles River, was clarified in the protocol subsequent to study initiation.

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1,2-Cyclohexanedicarboxylic Acid, 1-(phenylmethyl) ester, ester with 2,2,4-trimethyl, 1,3-petanediol mono(2-methyl propanoate)

EC Number:

950-347-5

Cas Number:

1661012-65-2

Molecular formula:

C27H40O6

IUPAC Name:

1,2-Cyclohexanedicarboxylic Acid, 1-(phenylmethyl) ester, ester with 2,2,4-trimethyl, 1,3-petanediol mono(2-methyl propanoate)

Test material form:

liquid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test Animals
Animals were received from Charles River, Stone Ridge, NY on 19 May 2015 and 09 Jun 2015. Following an acclimation period of at least five days, five healthy male and five healthy, non-pregnant and nulliparous female Sprague-Dawley rats were randomly assigned to the treatment groups.

The animals were born 23 Mar 2015, 13 Apr 2015 and 20 Apr 2015. The pretest body weight range was 243 - 285 g for males and 204 - 223 g for females.

The animals were identified by an indelible body mark and individually housed in suspended wire cages. Absorbent paper bedding was placed beneath the cages and changed at least three times per week. Fresh PMI Rat Chow (Diet #5012) was provided daily. Water was available ad libitum. The animal room, reserved exclusively for rats on acute tests, was temperature controlled, had a 12-hour light/dark cycle, and was kept clean and vermin free.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Site Preparation
The day prior to application of the test article, the dorsal area of the trunk of each animal was clipped free of hair. The clipped area began at the shoulders and extended to the hipbone and half way down the flank of each side of the animal. The prepared site was approximately 10% of the body surface and remained intact.

Dosing
A single dose of the test article was applied to the prepared site, over a porous gauze dressing at a dose level of 2000 mg/kg. The dose was based on the sample weight as calculated from the specific gravity. The torso was covered with a piece of porous dressing (semi-occlusive) to retain the gauze patch and was secured with non-irritating tape. The test article remained in contact with the skin for 24 hours at which time the wrappings were removed. Residual test article was removed by gently washing with distilled water.

Duration of exposure:

24 hours

Doses:

Test article applied unchanged (100% concentration)

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Type and Frequency of Observations
In vivo
- The test sites were scored for dermal irritation at 24 hours postdose and on Day 14 using the numerical Draize scoring code below. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction. Additional signs were described.

Erythema and Eschar
No erythema 0
Very slight erythema (barely perceptible) 1
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Edema
No edema 0
Very slight edema (barely perceptible) 1
Slight edema (edges of area well-defined by definite raising) 2
Moderate edema (raised approximately 1.0 mm) 3
Severe edema (raised more than 1.0 mm, extending beyond the area of exposure) 4

Type and Frequency of Observations (continued)
The animals were observed 1and 4 hours postdose and once daily for 14 days for toxicity and pharmacological effects.

Body weights were recorded pretest, weekly and at termination.

Post Mortem
– All animals were humanely sacrificed using CO2 following study termination and were examined for gross pathology.

Analysis of Data
An estimate of the LD50 was made based on mortality occurring during the study. Analysis was conducted according to the current Globally Harmonized System of Classification and Labeling of Chemicals.

Statistics:

None - N/A

Results and discussion

Preliminary study:

None

Effect levels

Key result

Sex:

male/female

Dose descriptor:

approximate LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat. (total fraction)

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

All five male and all five female rats survived the 2000 mg/kg 24-hour dermal exposure.

Clinical signs:

other: Dermal Observations At 24 hours post-dosing, erythema was absent to well-defined and edema was absent. By Day 14, erythema was absent to very slight and edema was absent; flaking skin was observed. Systemic Observations Abnormal physical signs including

Gross pathology:

Gross necropsy of all animals revealed no observable abnormalities.

Any other information on results incl. tables

Table 1. Body Weights and Dose Volume
Animal No.	Sex	Dose Volume (mL)	Body Weight (g)
			Day 0	Day 7	Day 14
1	Male	0.43	243	273	307
2	Male	0.45	252	288	326
3	Male	0.46	258	295	331
4	Male	0.46	257	285	314
5	Male	0.51	285	334	385
Mean			259	295	333
S.D.			15.7	23.2	30.8
#			5	5	5
6	Female	0.37	207	211	213
7	Female	0.40	223	238	244
8	Female	0.39	217	234	252
9	Female	0.37	204	219	224
10	Female	0.39	218	231	234
Mean			214	227	233
S.D.			8.0	11.2	15.5
#			5	5	5

Table 2. Dermal Observations
Time Periods		Animal Number and Sex
		Male					Female
		1	2	3	4	5	6	7	8	9	10
24-hour	Erythema	0	0	0	0	0	0	1	2	2	1
	Edema	0	0	0	0	0	0	0	0	0	0
Day 14	Erythema	0	0	0	0	0	0	0	0	1, f	0
	Edema	0	0	0	0	0	0	0	0	0	0

f = flaking skin

Table 3. Necropsy Observations
Animal Number	1	2	3	4	5	6	7	8	9	10
Sex	Male					Female
Death (D) / Sacrifice (S)	S	S	S	S	S	S	S	S	S	S
Observation
Appeared normal / No findings	X	X	X	X	X	X	X	X	X	X

X: Observed

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Based on the results observed, the dermal LD50 of Santicizer P1700 was determined to be greater than 2000 mg/Kg of body weight in rats. According to GHS Classification, Santicizer P1700 meets the criteria to be classified as Acute Toxic Category 5.

Executive summary:

A key EPA Guideline OPPTS 870.1200 study was conducted to determine the potential for toxicity of the test material (Santicizer P1700) when applied dermally. Five male and five female Sprague-Dawley rats were dosed dermally with Santicizer P1700 at 2000 mg/Kg of body weight. The test material was kept in contact with the skin for 24 hours and dermal responses recorded at 24 hours post exposure and on Day 14. Animals were observed for toxicity and pharmacological effects at 1 and 4 hours post exposure and once daily for 14 days. Body weights were recorded pretest, weekly and at termination and all animals were examined for gross pathology.

 

No mortality was observed through the study period and abnormal physical signs such as chromorhinorrhea, wetness and soiling of the anogenital area, erythema on the ears and partially chewed food on the cage pan liner were observed. At 24 hours post exposure, erythema was absent to well-defined and edema was absent. By Day 14, erythema was absent to very slight and edema was absent; flaking skin was observed. All animals in both sexes gained body weight by study termination and gross necropsy revealed no remarkable findings.

 

Based on the results observed, the dermal LD₅₀ of Santicizer P1700 was determined to be greater than 2000 mg/Kg of body weight in rats. According to GHS Classification, Santicizer P1700 meets the criteria to be classified as Acute Toxic Category 5.