Santicizer P1400

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-08-11 to 2011-03-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

not specified

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

Scientifically valid and acceptable delayed contact dermal sensitization test in guinea pigs. The study was conducted in the US in 2011.

Test material

Test material information

Constituent 1

Reference substance name:

benzyl butyl cis-cyclohexane-1,2-dicarboxylate

Cas Number:

1931129-39-3

Molecular formula:

C19H26O4

IUPAC Name:

benzyl butyl cis-cyclohexane-1,2-dicarboxylate

Test material form:

liquid

Specific details on test material used for the study:

Label Identity: Santicizer P-1400
Batch No: RP-620
Supplied by: Ferro Corporation
Data received: 08/19/2010
Storage: Room temperature and humidity
Description: Clear yellow liquid
Sample preparation: Induction and challange - The test article was used as received.
Screen: Used as received (100% concentration) and was dissolved in ethanol and diluted to appropriate concentrations (10, 25 and 50) with distilled water (opaque liquid).

TEST DATES:
Study Initiation: 08 November 2010
Experimental Start Date: 10 November 2010
Experimental Term Date: 16 December 2010
Draft Report Signed: 06 January 2011
Final Report Signed: 03 March 2011

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Elm Hill Breeding Labs, Inc., Chelmsford, MA
- Age at study initiation:
- Weight at study initiation: Male: 277-474 g - Female: 300-465 g
- Housing: suspended wire cages
- Diet (e.g. ad libitum): PMI Guinea Pig Chow - ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): This parameter deviated from the standard protocol. This did not have any effect on the study animals and the integrity of the study.
- Humidity (%): This parameter deviated from the standard protocol. This did not have any effect on the study animals and the integrity of the study.
- Air changes (per hr): no info
- Photoperiod (hrs dark / hrs light): 12 hr light and 12 hr dark

IN-LIFE DATES: From: 11/10/10 to12/06/11

Study design: in vivo (non-LLNA)

Induction

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

0.4 ml

Day(s)/duration:

6 hours

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

0.4 mL

Day(s)/duration:

6 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10 males and 10 females ( 0.4 mL concentration)
5 males and 5 females (control)

Details on study design:

Site preparation:
Site 1 and 2: tests
Site 3 and 4: alternative site in the event that severe irritation was noted

SCREEN:
Each animal received 4 concentration of the test article, one/site. Each concentration (10, 25, 50, 100%) was placed in 25 mm Hilltop Chamber contained a cotton pad. The sites were covered with a strip of rubber dental dam to cover the treated sites and wrapped with tape to provide occlusion. After 6 hours the dams and test article were removed and the sites cleaned with distilled water and dried with a soft towel.

INDUCTION EXPOSURE
Group 1 (10 males and 10 females) was treated with 0.4 ml of the test article (100%). The dose was applied to the left shoulder area (site1) using 25 mm Hilltop Chamber contained a cotton pad in order to facilitate the contact of the liquid test article with the site. The chamber was covered with a strip of rubber dental dam and the torso was wrapped with no-irritant tape to provide occlusion. After 6 hours, the dam and test articles were removed and the site was cleaned with ethanol.
This procedure was performed one/week for three weeks. Group 2 (5 males and 5 females) were untreated for three weeks induction and served as negative control.

CHALLENGE EXPOSURE
After 14 days from the last induction, the animals of Group 1 and 2 were challanged using the same dosing procedure as in the induction phase. 100% was chosen as the highest non-irritating concentration for the challange (based on the screen results). The doses were applied to Site 3, lower left dorsal area.

Challenge controls:

5 males and 5 females

Positive control substance(s):

yes

Remarks:

HCA

Results and discussion

Positive control results:

Hexylcinnamaldehyde, HCA: Incidence index : 24 hr: 0.05; 48 hr: 0.32. Severity index 24 hr: 0.18; 48hr: 0.39 - Historic data retested every 6 months in the lab.

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

negative control

Dose level:

100%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No abnormal systemic observations

Remarks on result:

other: Naive to induction

Results 2

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

negative control

Dose level:

100%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No systemic abnormalities

Remarks on result:

other: Naive to induction

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

other: First induction

Results 4

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

Mo systemic abnormalities

Remarks on result:

other: First induction

Results 5

Key result

Reading:

2nd reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

other: Second induction

Results 6

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

other: Second induction

Results 7

Key result

Reading:

other:

Hours after challenge:

24

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

other: Third induction

Results 8

Key result

Reading:

other:

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

other: Third induction

Results 9

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

no indication of skin sensitisation

Results 10

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No abnormal systemic observations

Remarks on result:

no indication of skin sensitisation

Results 11

Key result

Reading:

other: HCA Positive control data retested every 6 months

Hours after challenge:

24

Group:

positive control

Dose level:

HCA (85%)

Remarks on result:

other: Incidence index 0.05

Remarks:

Severity index 0.18

Results 12

Key result

Reading:

other: HCA Positive control restested every 6 months

Hours after challenge:

48

Group:

positive control

Dose level:

HCA (85%)

Remarks on result:

other: Incidence index 0.32

Remarks:

Severity index 0.39

Any other information on results incl. tables

Table 1. Results
Challenge	Incidence Index		Severity Index
	24 hours	48 hours	24 hours	48 hours
Santicizer P1400	0	0	0	0
Naïve Control	0	0	0	0
Historical Positive Control^a
HCA Positive Control	0.05	0.32	0.18	0.39
Naïve Control	0	0	0	0

^aHistorical Positive Control data (MB 10-19286.06) attached in Appendix A of the study report.

Table 2. Dermal Observations and Body Weights (g) - Screen
Animal No. / sex	Pretest Body Weight (g)	Site	Concentration (%)	Erythema
				24 hours	48 hours
D4195 / Male	464	1	100	0.5	0
		2	50	0	0
		3	25	0	0
		4	10	0	0
D4196 / Male	474	1	100	0.5	0
		2	50	0	0
		3	25	0	0
		4	10	0.5	0
D4197 / Female	465	1	100	0	0
		2	50	0	0
		3	25	0	0
		4	10	0	0
D4198 / Female	410	1	100	0	0
		2	50	0	0
		3	25	0	0
		4	10	0	0

Table 3. Dermal Observations: Group 1 Post Patch Removal (Induction) – 0.4 mL (100% Concentration)
Animal No. / Sex	Induction 1, Erythema Site# 1		Induction 2, Erythema Site# 1		Induction 3, Erythema Site# 1
	24 hours	48 hours	24 hours	48 hours	24 hours	48 hours
D4271 / Male	0	0	0	0	0	0
D4272 / Male	0	0	0	0	0	0
D4273 / Male	0	0	0	0	0	0
D4274 / Male	0	0	0	0	0	0
D4275 / Male	0	0	0	0	0	0
D4276 / Male	0	0	0	0	0	0
D4277 / Male	0	0	0	0	0	0
D4278 / Male	0	0	0	0	0	0
D4279 / Male	0	0	0	0	0	0
D4280 / Male	0	0	0	0	0	0
D4281 / Female	0	0	0	0	0	0
D4282 / Female	0	0	0	0	0	0
D4283 / Female	0	0	0	0	0	0
D4284 / Female	0	0	0	0	0	0
D4285 / Female	0	0	0	0	0	0
D4286 / Female	0	0	0	0	0	0
D4287 / Female	0	0	0	0	0	0
D4288 / Female	0	0	0	0	0	0
D4289 / Female	0	0	0	0	0	0
D4290 / Female	0	0	0	0	0	0

 

Table 4. Body Weight (Group 1): Induction
Animal No. / sex	Pretest (g)	Term (g)
D4271 / Male	288	520
D4272 / Male	303	489
D4273 / Male	277	528
D4274 / Male	293	529
D4275 / Male	307	541
D4276 / Male	311	513
D4277 / Male	346	645
D4278 / Male	294	520
D4279 / Male	315	588
D4280 / Male	289	515
Mean	302	539
S.D.	19.3	45.1
#	10	10
D4281 / Female	357	567
D4282 / Female	314	492
D4283 / Female	317	433
D4284 / Female	308	482
D4285 / Female	312	514
D4286 / Female	316	494
D4287 / Female	300	523
D4288 / Female	354	549
D4289 / Female	305	451
D4290 / Female	318	416
Mean	320	492
S.D.	19.5	48.7
#	10	10

 

Table 5. Dermal Observations: Groups 1 and 2 Post Patch Removal (Challenge)
Animal No. / sex	Challenge Erythema (100% Concentration) Site# 3		Animal No. / sex	Challenge Erythema (Naïve Control) Site# 3
	24 hours	48 hours		24 hours	48 hours
D4271 / Male	0	0	D4291 / Male	0	0
D4272 / Male	0	0	D4292 / Male	0	0
D4273 / Male	0	0	D4293 / Male	0	0
D4274 / Male	0	0	D4294 / Male	0	0
D4275 / Male	0	0	D4295 / Male	0	0
D4276 / Male	0	0
D4277 / Male	0	0
D4278 / Male	0	0
D4279 / Male	0	0
D4280 / Male	0	0
D4281 / Female	0	0	D4296 / Female	0	0
D4282 / Female	0	0	D4276 / Female	0	0
D4283 / Female	0	0	D4298 / Female	0	0
D4284 / Female	0	0	D4299 / Female	0	0
D4285 / Female	0	0	D4300 / Female	0	0
D4286 / Female	0	0
D4287 / Female	0	0
D4288 / Female	0	0
D4289 / Female	0	0
D4290 / Female	0	0

 

Table 6. Body Weight (Naïve Controls - Group 2): Challenge
Animal No. / sex	Pretest (g)	Term (g)
D4291 / Male	319	640
D4292 / Male	312	558
D4293 / Male	294	535
D4294 / Male	310	581
D4295 / Male	349	648
Mean	317	592
S.D.	20.2	49.9
#	5	5
D4296 / Female	356	487
D4276 / Female	352	535
D4298 / Female	326	542
D4299 / Female	308	438
D4300 / Female	316	424
Mean	332	485
S.D.	21.5	54.0
#	5	5

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Not sensitising

Conclusions:

Based on the results observed, Santicizer P1400 was determined to not be a dermal sensitizer under the conditions of this delayed contact dermal sensitization test in guinea pigs.

Executive summary:

In a key EPA OPPTS Guideline 870.2600 delayed contact dermal sensitization study, the potential of the test material (Santicizer P1400) to promote skin sensitization reactions following repeated applications was evaluated in guinea pigs using the Buehler method.

 

Seventeen healthy male and female Hartley Albino guinea pigs were utilized in the study. Four guinea pigs were used to screen for the highest non-irritating concentration which produced scores no more severe than two scores 0.5 and two scores of 0. Based on the results of this screen, 100% was selected as the highest non-irritating concentration for the challenge and administered to two groups of guinea pigs (Group 1 and Group 2) two weeks subsequent to the third induction.

 

Group 1 (10 males and 10 females) was induced with the test material at a concentration of 100%. Group 2 (5 males and 5 females) animals were not induced and served as the naïve control. For the induction phase, a dose of 0.4 mL (100% concentration) was applied to the left shoulder area (25 mm²area) of 10 male and 10 females guinea pigs in Group1. After application, the torso was wrapped with non-irritant tape to provide occlusion. The site was exposed for 6 hours and then cleaned with ethanol. Group 1 animals received three topical induction applications, one per week for three weeks. Skin reactions of all animals were recorded 24 and 48 hours following patch removal. 14 days after the last treatment, animals of Group 1 and 2 were challenged following the same procedure as used for the induction phase. The dose was applied on the lower left dorsal area. Body weights were recorded pretest and at termination and all animals were observed once per day for mortality and toxicity. 85% hexylcinnemaldehyde (HCA) was the historical positive control with its toxicity confirmed in the laboratory every six months.

 

No mortality was observed and body weight changes in all animals were normal through the study period. Treated as well as control animals appeared normal through the observation period. Erythema was absent at induction as well as at challenge in both the induced as well as naïve control groups.

 

Based on the results observed, Santicizer P1400 was determined to not be a dermal sensitizer under the conditions of this delayed contact dermal sensitization test in guinea pigs.