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Description of key information

An Open Epicutaneous Test (OET) was performed on guinea pigs  to assess the skin sensitization potential of test chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7).On day 1 during induction, 0.1 ml of the alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was applied at concentrations of 100%, 30%, 10%, 3%, 1%, or 0.3% in vehicle to an area measuring 8 cm2 on the clipped flank skin of the guinea pigs. The applications are repeated daily for 3 weeks or done 5 times weekly during 4 weeks, usually on the same skin sites. The application sites were left uncovered and the reactions, if continuous daily applications were performed, can be read 24 h after each application, or at the end of each week. To determine whether or not contact sensitization was induced, all groups of guinea pigs previously treated for 21 days, as well as 10 untreated, or only pretreated with the vehicle, controls are tested on days 21 and 35 on the contralateral flank with the test material. This test was performed by applying with a pipette 0.025 ml of each concentration to skin areas measuring 2 cm2. The reactions were read after 24,48 and/or 72h. It was observed that none of the guinea pigs induced contact sensitization at challenge concentration of 6%.Thus the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered to be not sensitizing on skin of guinea pigs at concentration of 6% in an Open Epicutaneous Test (OET).

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed journal
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
An Open Epicutaneous Test (OET) was performed on guinea pigs to assess the skin sensitization potential of test chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7).
GLP compliance:
not specified
Type of study:
open epicutaneous test
Justification for non-LLNA method:
not specified
Specific details on test material used for the study:
- Name of test material (as cited in study report): alpha-Amyl cinnamaldehyde
- Molecular formula: C14H18O
- Molecular weight: 202.2952 g/mol
- Substance type: organic
- Physical state: liquid
-Smiles: c1(\C=C(\CCCCC)C=O)ccccc1
-InChI: 1S/C14H18O/c1-2-3-5-10-14(12-15)11-13-8-6-4-7-9-13/h4,6-9,11-12H,2-3,5,10H2,1H3/b14-11-
Species:
guinea pig
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
- Weight at study initiation: 300-450g
Route:
epicutaneous, open
Vehicle:
other: ethanol, acetone, H20, petroleum, PEG and/or other suitable vehicles
Concentration / amount:
Concentration:100%, 30%, 10%, 3%, 1%, or 0.3%
Amount: 0.1ml
Day(s)/duration:
3 weeks (0-21 days)
Route:
epicutaneous, open
Vehicle:
other: ethanol, acetone, H20, petroleum, PEG and/or other suitable vehicles
Concentration / amount:
on days 21 and 35
Day(s)/duration:
Concentration:6% Amount: 0.025ml
Adequacy of challenge:
not specified
No. of animals per dose:
6-8 guinea pigs
Details on study design:
RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 21
- Exposure period:24 hours
- Test groups:6-8 guinea pigs
- Control group:3 guinea pigs
- Site: an area measuring 8 cm2 on the clipped flank skin of the guinea pigs
- Frequency of applications: The applications are repeated daily for 3 weeks or done 5 times weekly during 4 weeks
- Duration: 21 days (3 weeks)
- Concentrations: 100%, 30%, 10%, 3%, 1%, or 0.3% in vehicle.
Amount: 0.1ml

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: on days 21 and 35
- Exposure period:24 hours
- Test groups: 6-8 guinea pigs
- Control group: 3 guinea pigs
- Site: contralateral flank measuring 2 cm2
- Concentrations: Concentration:6%
Amount: 0.025ml
- Evaluation (hr after challenge): 24,48 and/or 72h.
Challenge controls:
Yes concurrent vehicle.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
6%
No. with + reactions:
0
Total no. in group:
8
Clinical observations:
No known signs of skin sensitization were observed.
Remarks on result:
no indication of skin sensitisation
Cellular proliferation data / Observations:
No known signs of skin sensitization were observed.

1 day before starting the induction procedure, the threshold irritating concentration of the test material is estimated on the guinea pigs subsequently used for the experimental group, A single application of 0.025 ml of each test concentration (e.g, 100, 30, 10 and 3%) is simultaneously performed on one of the areas measuring 2 cm2 of the flank skin previously clipped and marked with a circular stamp. Reactions are read 24 h after the application of the test material. The minimal irritating and the maximal nonirritating concentrations are determined by an all-or-none criterion. The minimal irritating concentration is defined as the lowest one causing skin irritation. The maximal nonirritating concentration is defined as the highest one not causing macroscopic skin reactions in any of the animals. The estimation of the threshold concentration (minimal irritating and maximal nonirritating concentration) is essential for the evaluation of the allergenic capacity of the test material.

Interpretation of results:
other: Not sensitizing
Conclusions:
The test material alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered to be not sensitizing on skin of guinea pigs at concentration of 8% in an Open Epicutaneous Test (OET).
Executive summary:

An Open Epicutaneous Test (OET) was performed on guinea pigs to assess the skin sensitization potential of test chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7).

 

On day 1 during induction, 0.1 ml of the alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was applied at concentrations of 100%, 30%, 10%, 3%, 1%, or 0.3% in vehicle to an area measuring 8 cm2 on the clipped flank skin of the guinea pigs. The applications are repeated daily for 3 weeks or done 5 times weekly during 4 weeks, usually on the same skin sites. The application sites were left uncovered and the reactions, if continuous daily applications were performed, can be read 24 h after each application, or at the end of each week.

 

To determine whether or not contact sensitization was induced, all groups of guinea pigs previously treated for 21 days, as well as 10 untreated, or only pretreated with the vehicle, controls are tested on days 21 and 35 on the contralateral flank with the test material. This test was performed by applying with a pipette 0.025 ml of each concentration to skin areas measuring 2 cm2. The reactions were read after 24,48 and/or 72h.

 

It was observed that none of the guinea pigs induced contact sensitization at challenge concentration of 6%.Thus the chemicalalpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered to be not sensitizing on skin of guinea pigs at concentration of 6% in an Open Epicutaneous Test (OET).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

SKIN SENSITIZATION:

Various studieshas been investigated for the test chemicalalpha-Amyl cinnamaldehyde (CAS No: 122-40-7)to observe the potential for skin sensitization to a greater or lesser extent. The studies are based on in vivo experiments in rabbits, guinea pigsand humans for target chemical-Amyl cinnamaldehyde (CAS No: 122-40-7) which have beensummarized as below;

An Open Epicutaneous Test (OET) was performed by G. Klecak (1985)on guinea pigs  to assess the skin sensitization potential of test chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7).On day 1 during induction, 0.1 ml of the alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was applied at concentrations of 100%, 30%, 10%, 3%, 1%, or 0.3% in vehicle to an area measuring 8 cm2 on the clipped flank skin of the guinea pigs. The applications are repeated daily for 3 weeks or done 5 times weekly during 4 weeks, usually on the same skin sites. The application sites were left uncovered and the reactions, if continuous daily applications were performed, can be read 24 h after each application, or at the end of each week. To determine whether or not contact sensitization was induced, all groups of guinea pigs previously treated for 21 days, as well as 10 untreated, or only pretreated with the vehicle, controls are tested on days 21 and 35 on the contralateral flank with the test material. This test was performed by applying with a pipette 0.025 ml of each concentration to skin areas measuring 2 cm2. The reactions were read after 24,48 and/or 72h. It was observed that none of the guinea pigs induced contact sensitization at challenge concentration of 6%.Thus the chemicalalpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered to be not sensitizing on skin of guinea pigs at concentration of 6% in an Open Epicutaneous Test (OET).

 

The G. Klecak , H. Geleick and J. R. Frey (1985) carried out two screening test namely Draize test (DT) and maximisation test (MT) of chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) in Himalayan white-spotted guinea pigs as follows;

In theDraize test, the 6-8 guinea pigs received a dose of 0.05 ml of a 0.1 % solution of the chemical tested in isotonic saline intradermally on day 0 and further doses of 0.1 ml each were injected on 9 alternate days (total dose = 0.95 rag). The treated animals and untreated controls were challenged intradermally with 0.05 ml of a 0.1 per cent solution on days 35 and 49. The evaluation criterion was the mean diameter of the popular reactions. None of the treated animals showed positive skin reactions.

In Guinea pig maximisation testduring induction, on day 0the animals were injected intradermally with 0.1 ml of a   5% solution of the compound tested, with 0.1 ml of a 5 % emulsion of the same compound in Freund's complete adjuvant (FCA) and with 0.1 ml of FCA alone. On day 8 the animals were induced epicutaneously with 250 mf of 25% test compound in dissolved in petrolatum on clipped skin area of the neck under occlusion for 2 days. On day 21, animals were challenged with the compound at a subirritant concentration in petrolatum was applied to the flank for 24 h. The reactions were read 24 and 48 h after removing the patch.

Since no skin reactions were observedin both the tests, the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7)was considered to be not sensitizingto the skin of Himalayan white-spotted guinea pigs.

 

G. Klecak (1985) performed another FREUND'S COMPLETE ADJUVANT (FCAT) in Himalayan white-spotted guinea pigs (male and female) of alpha-Amyl cinnamaldehyde (CAS No: 122-40-7).In this test, two groups of 10-20 guinea pigs each were involved: (a) the experimental group, and (b) the control group. Additionally, 4 guinea pigs were used for determination of the primary irritating concentration of the test substance so that nonirritating concentrations may be chosen for challenge For induction, the scapula region of test and control animals is shaved. During days 1-9 the animals of the experimental group are treated with 0.1 ml of the test material in FCA intradermally into the scapula region three times (days I, 5 and 9) on an area of 6 x 2 cm. The animals of the control group are treated with 0.1 ml of FCA only.To determine whether or not contact allergy was induced, both animal groups, the experimental and the control one, are challenged on days 21 and 35 on the contralateral flanks with the same compound at the minimal irritating and some lower nonirritating concentrations. The tests are performed by applying with a pipette 0.025 ml of each concentration to skin areas measuring 2 cm2 The reactions are read 24, 48 and 72 h after application.No known signs of skin reactions were observed during 72 hours observation period. Thus the chemicalalpha-Amyl cinnamaldehyde (CAS No: 122-40-7)was considered to be not sensitizingto the skin of Himalayan white-spotted guinea pigs.

 

 

The above results were further supported by Human maximisation test reported by D. L. J. OPDYKE (1973)and the MAK Collection for Occupational Health and Safety (2014) of alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) on 25 volunteers in which each subject received 6% of alpha-Amyl cinnamaldehyde and then observed for skin reactions. The chemical did not lead to sensitization in any of 25 persons. Therefore alpha-Amyl cinnamaldehyde (CAS No: 122-40-7)was considered to be not sensitizing.

 

 

A. C. DE GROOT, A.M. J. VANDER KLEY, D.P. BRUYNZEEL, M. M. H. M. MEINARDI G. SMEENK, TH. VAN JOOST AND S. PAVEL (1993) carried out a patch test of alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) in 677 patients. The patients were exposed to 5% of alpha-Amyl cinnamaldehyde in petrolatum and then observed for skin reactions. Positive skin reactions were observed in only 3 patients out of 677 patients. Thus the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered as not sensitizing.

 

J. D. Wilkinson, K. Andersen, J. Camarasa, G. Ducombs, P. Frosch, A. Lahti, T.Menne, R.J.G.Rycroft, and I.White (1989) performed a patch test on 78 patients to establish the frequency of positive reactions of test chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7). A patch of 2% of alpha-Amyl cinnamaldehyde in petrolatum was applied dermally on skin of each patient and then observed for skin reactions. Positive reactions were observed in 2 patients out of 78 patients. Thus the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered as not sensitizing.

 

Temesvári, E., Németh, I., Baló-Banga, M. J., Husz, S., Kohánka, V., Somos, Z., Judák, R., Remenyik, É. V. A., Szegedi, A., Nebenführer, L., Mészáros, C. and Horváth, A. (2002) conducted patch test of chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) to assess fragrance sensitization in the Hungarian population in the years 1998 and 1999. A patch of 1% of alpha-Amyl cinnamaldehyde in petrolatum was applied epicutaneously on skin of 116 patients for 24 hours exposure period and then observed for skin reactions.The skin lesions were evaluated at 20, 40, and 0 min of testing, and at 24, 48 and 72h after a 24-h application. Skin lesions were observed in only 5% of the 116 patients. Thus the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered as not sensitizing.

B. SANTUCCI, A. CRISTAUDO, C. CANNISTRACI AND M. PICARDO (1987) and the MAK Collection for Occupational Health and Safety (2014) carried out two patch studies of chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) as follows;

The first patch test of chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was carried out in 63 patients to evaluate the incidence of contact dermatitis from November 1983 to June 1984. The patches were applied on 2 Sides of the upper back at concentration of 3% in petrolatum under occlusive condition using Finn Chambers on Scanpor. Skin reactions were evaluated at 48, 72 and 96 h, according to the ICDRG scale; the last reading was taken as defimtive. Since the chemical induce contact allergy in 3out of 63 patients, the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considerednot sensitizing.

The second patch test of chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was carried out in 54 patients from October 1984 to June 1985 by the same procedure as mentioned in previous patch test. Since the chemical induce contact allergy in 1out of 54 patients, the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered as not sensitizing.

 

Another study conducted by Gerberick GF, Ryan CA, Kern PS, Schlatter H, Dearman RJ, Kimber I, Patlewicz GY, Basketter DA (2005) by using the Mouse Local Lymphnode Assay on chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7).The LLNA was conducted on groups of CBA mice (7-12 weeks of age) by mean of topical application of chemical on the dorsum of both ears at a dose of 25µl or to an equal volume of relevant vehicle (Acetic acid in olive oil (4:1))only. Treatment was performed daily for 3 consecutive days. Five days after initiation of exposure all mice were injected via the tail vein with 250µl of PBS containing 20µCi of tritiatied thymidine. The mice were sacrificed 5 hours later, and draining the auricular lymph nodes were excised and pooled for each experimental group or each individual animal. The incorporation of tritiated thymidine measured by β-scintillation counting and was reported in disintegrations /minute. An SI was calculated for each chemical group as the ratio of disintegrations/minute of the treated group to the disintegrations/minute of the concurrent vehicle control group. A substance was classified skin sensitizer, if at one or more than one concentrations, it induced a three-fold or greater increase in local lymph node proliferative activity when treated with the concurrent vehicle treated controls (SI ≥3).The approach to estimation of the relative skin sensitization potential is based on the mathematical estimation of the concentration of chemical necessary to obtain a threshold positive response (SI = 3); this is termed as the EC3 value.The relative potency index of alpha-Amyl cinnamaldehyde was estimated to be 11.Based on the relative potency index alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) was considered as weak sensitizer.

Jere D. Guin, M.D., and Philip Haffley, Ph.D. Kokomo, IN. (1983) carried out patch ofα-Amylcinnamic aldehyde (CAS No: 122-40-7) in humans. Fourteen humans were used for patch testing at concentration of 5%α-Amylcinnamic aldehyde applied on the back of human subjects for 3 year period. A review of patch test results for a 3-year period revealed that fourteen patients had reactions of α-Amylcinnamic aldehyde. Of these 14 humans, ten showed mild reaction (+) reactions may or may not be clinically significant. The concentration was well below that reported to be nonirritating to human skin. Strong (++) patch test reactions to perfume materials in petrolatum applied to the back can be associated with clinical dermatitis of the face and groin that is caused by very dilute concentrations of fragrance ingredients. The only male patient with a strongly positive (+ + +) patch test reaction had numerous sensitivities that were probably causally related to his condition, but what part fragrances played is not clear because his condition cleared promptly with simple topical treatment, together with avoidance of all potential problem products. The chronic vulvar dermatitis found in all three female patients with strongly positive patch test reactions could be the cause or the result of sensitization to α-Amylcinnamic aldehyde.On the basis of positive indication of skin sensitizationin humans by patch test it was concluded that the α-Amylcinnamic aldehyde alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) is sensitizing to the skin.

 

 Another Local lymph node assay (LLNA) was reported by European Commission Health & Consumers (2012) of alpha-Amyl cinnamaldehyde (CAS No: 122-40-7). During the assay, 4 animals were exposed to alpha-Amyl cinnamaldehyde at a concentration of 1.0, 2.5, 5.0, 10.0, and 25.0%. Vehicle used was 1:3 (ethanol: diethyl phthalate). On the basis of observations and EC3 valuei.e. 7.6 obtained from the assay it was concluded that thealpha-Amyl cinnamaldehyde (CAS No: 122-40-7)was moderately sensitizing.

 

 

Although positive skin reactions were observed in few studies but the potential of skin sensitization was not enough to classify the chemical as skin sensitizer. Thus on the basis of available dataand majority of results obtained in various studies, the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) can considered as not sensitizing.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The skin sensitization potential of test substance alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) were observed in various studies. On the basis of available data and majority of results obtained in various studies, the chemical alpha-Amyl cinnamaldehyde (CAS No: 122-40-7) can considered as not sensitizing and hence can be classified as non skin sensitizer.